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REVIEWS Electrical synapses and their functional interactions with chemical synapses Alberto E. Pereda Abstract | Brain function relies on the ability of neurons to communicate with each other. Interneuronal communication primarily takes place at synapses, where information from one neuron is rapidly conveyed to a second neuron. There are two main modalities of synaptic transmission: chemical and electrical. Far from functioning independently and serving unrelated functions, mounting evidence indicates that these two modalities of synaptic transmission closely interact, both during development and in the adult brain. Rather than conceiving synaptic transmission as either chemical or electrical, this article emphasizes the notion that synaptic transmission is both chemical and electrical, and that interactions between these two forms of interneuronal communication might be required for normal brain development and function. Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA. e‑mail: alberto.pereda@ einstein.yu.edu doi:10.1038/nrn3708 Published online 12 March 2014 Communication between neurons is required for brain function, and the quality of such communication enables hardwired neural networks to act in a dynamic fashion. Functional interactions between neurons occur at anatomically identifiable cellular regions called synapses. Although the nature of synaptic transmission has been an area of enormous controversy (BOX 1), two main modalities of synaptic transmission — namely, chemical and electrical — are now recognized. At chemical synapses, information is transferred through the release of a neurotransmitter from one neuron and detection of the neurotransmitter by an adjacent cell1, whereas in electrical synapses, the cytoplasm of adjacent cells is directly connected by clusters of intercellular channels called gap junctions2. Although cellular specializations for both forms of transmission can be found at various neuronal sites (dendrites, somata and axons), chemical transmission normally occurs between synaptic terminals from axons and the dendrite or soma of a second neuron, muscle fibre or gland cell. The presence of these two modalities of synaptic transmission does not exclude the possibility that brain cells could also communicate by alternative mechanisms, such as volume transmission (diffusion of neurotransmitters through the extracellular space to reach remote target cells)3 and the generation of electrical fields that are capable of influencing the excitability of nearby neurons4. Electrical and chemical synapses are now known to coexist in most organisms and brain structures, but details of the properties and distribution of these two modalities of transmission are still emerging. Most research efforts have focused on exploring the mechanisms of chemical transmission, and considerably less is known about those underlying electrical transmission. It was thought that electrical synapses were more abundant in invertebrates and cold-blooded vertebrates than in mammals. However, a wealth of data now indicate that electrical synapses are widely distributed in the mammalian brain5. In addition to the retina, inferior olive and olfactory bulb, structures in which electrical transmission has been known to occur for some time5, electrical synapses have been found in disparate regions of the mammalian CNS and have been shown to constitute a distinct phenotypic feature of inhibitory interneurons in general6–8. Perhaps because the appreciation of each form of synaptic transmission occurred in a sequential manner (at least in mammals), there is a widespread notion that electrical and chemical synapses operate independently. This article reviews the evidence that chemical and electrical synapses functionally interact during development and in adulthood. Rather than being a comprehensive review of the experimental evidence, the focus here is on a few examples that highlight such functional interactions. 250 | APRIL 2014 | VOLUME 15 www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS Box 1 | The debate over the nature of synaptic transmission S. Ramón y Cajal and C. Sherrington, the fathers of modern neuroscience, established that networks of multiple elementary units, called ‘neurons’, communicate with each other through functional specializations called ‘synapses’. Their seminal contributions were followed by a bitter debate over the nature of synaptic transmission: was it mediated by chemical or electrical signals? This controversy was known as ‘The War of the Soup and the Sparks’ (REF. 170). Although several researchers, most notably T. R. Elliott171 and later O. Loewi172, demonstrated the existence of neurotransmitters with actions on postsynaptic cells, there was still controversy over whether transmitter release could occur in a fraction of a millisecond, the synaptic ‘delay’ indirectly measured by Sherrington173. B. Katz and colleagues174 demonstrated that synaptic transmission at the frog neuromuscular junction was an electrically mediated, calcium-dependent form of transmitter release, which occurred within a fraction of a millisecond. Transmission by such a mechanism was also shown to occur in the CNS, leading to a general agreement that synaptic transmission was chemically mediated. However, in 1958, D. Potter communicated at a ‘Monday night fight’ of the Marine Biological Laboratory in Woods Hole (USA) (so called because of the contentious nature of the scientific exchanges) the striking properties of synaptic transmission in crayfish, which challenged all the criteria established for chemical transmission. Postsynaptic signals reproduced the time course of presynaptic signals, and transmission was bidirectional and, surprisingly, voltage-dependent. The findings provided the earliest evidence in support of the existence of electrical synaptic transmission175 and were soon followed by seminal studies in the teleost brain by M. V. L. Bennett and colleagues176, J. D. Robertson and colleagues177 and E. J. Furshpan178, in which physiological and ultrastructural analyses were combined. Their search for the anatomical basis of electrical transmission greatly contributed to the identification of the cellular structures that we know today as gap junctions. The more recent demonstration of the ubiquitous presence of electrical synapses in the mammalian brain led to the indisputable conclusion that chemical transmission and electrical transmission coexist in all nervous systems. Lateral excitation The ability of an excited neuron (or sensory afferent) to excite its neighbours. Although it reduces discrimination, lateral excitation greatly enhances input sensitivity. It is a less-appreciated property of sensory and cortical networks. Are chemical synapses more sophisticated? Chemical versus electrical synapses. From an evolutionary point of view, chemical communication between cells preceded electrical communication9. Communication with chemical signals occurs in unicellular organisms such as bacteria10, in which it mediates important phenomena such as quorum sensing 11 (the mechanism underlying the detection of bacterial population density). More-specific communication between individual cells requires complex cellular specializations between the interacting partners. The presence of such cellular specializations is not exclusive to neurons and is a characteristic that neuronal chemical and electrical synapses share with immunological synapses between a lymphocyte and an antigen-presenting cell12, which dynamically form to facilitate the transfer of information between these two cells. Chemical transmission requires sophisticated presynaptic molecular machinery that regulates neurotransmitter release in a probabilistic manner when an action potential invades the synaptic terminal1 (although at some contacts, release can also be proportional to changes in resting membrane potential13). Such transmission also requires a similarly complex postsynaptic molecular machinery, including ionotropic (ligand-gated ion channels) and metabotropic receptors (G protein-coupled receptors that act indirectly through a secondary messenger) (FIG. 1a), which are capable of detecting and translating messages into various postsynaptic events, ranging from changes in resting potential (‘synaptic potential’) to gene expression1. These features enable chemical synapses to adapt to diverse functional requirements. The mechanisms and general properties of chemical synaptic transmission have been extensively reviewed (see REF. 1 for a review). Electrical transmission between neurons is mediated by gap junctions2, which are clusters of intercellular channels that directly connect the interiors of two adjacent cells (FIG. 1b), enabling the bidirectional passage of electrical currents and small molecules (for example, calcium, cyclic AMP and inositol-1,4,5‑trisphosphate). Gap junction channels are formed by the docking of two hexameric connexin ‘hemichannels’ (also called ‘connexons’); that is, one from each adjacent cell14. Strikingly, although they assemble into almost identical structures, connexons in invertebrates and vertebrates are formed by two different multigene families of proteins: namely, connexins, which are unique to chordates, and a second family including innexins and pannexins, which are found in invertebrates, prechordates and chordates2,14 (note that in contrast to innexins, pannexins do not seem to form intercellular channels in tissues and thus are thought to operate only as unpaired hemichannels in vertebrates15). Such evolutionary convergence highlights the functional importance of gap junction intercellular communication. From a family of over 20 genes in mammals14, only a handful of connexins have been found to be expressed in vertebrate neurons, including connexin 36 (CX36; also known as GJD2), CX45 (also known as GJC1), CX50 (also known as GJA8), CX30.2 (also known as GJC3) and CX31.1 (also known as GJB5) (see REFS 16–18 for reviews). To date, the retina is the neuronal tissue in which the diversity of neuronal connexins has been most investigated (see REF. 19 for a review). Because of its widespread distribution, CX36 is considered to be the main ‘synaptic’ connexin in the mammalian brain20. However, although electrical transmission is dramatically reduced in CX36‑knockout mice, some level of electrical and tracer coupling can be detected in these animals21,22, suggesting that synaptic gap junctions may be composed of additional connexins. Several of the 25 members of the innexin family have been identified in invertebrate neurons, mainly in fly 23, leech24 and worm neurons25. Although electrical synapses can act, to some extent, in a metabotropic manner by allowing the passage of small metabolites (a critical property of electrical synapses during development; see below), they, unlike chemical synapses, lack the ability to amplify and transform presynaptic signals. However, this does not mean that electrical synapses are less sophisticated; their sophistication just relies on a different functional property — their bidirectionality — which enables them to coordinate the activity of large groups of interconnected neurons2. Based on this bidirectional and analogical (that is, they do not require an action potential) nature, electrical synapses are particularly efficient at detecting the coincidence of simultaneous subthreshold depolarizations within a group of coupled neurons, a phenomenon that increases neuronal excitability and promotes synchronous firing 22,26–29. Electrical synapses are also highly effective at mediating lateral excitation and increasing the sensitivity of sensory systems, as it NATURE REVIEWS | NEUROSCIENCE VOLUME 15 | APRIL 2014 | 251 © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS a Chemical synapse b Electrical synapse Action potential Ca2+ Action potential Presynaptic terminal Synaptic vesicle Gap junction channel Neurotransmitter Ionotropic receptor Metabotropic receptor Membrane potential Postsynaptic terminal Gene expression Coupling potential Biochemical cascades Figure 1 | The two main modalities of synaptic transmission. a | Chemical transmission requires sophisticated Nature Reviews | Neuroscience presynaptic molecular machinery that regulates neurotransmitter release in a probabilistic manner upon depolarization of the presynaptic terminal — in this case, by the arrival of an action potential — which leads to the activation of voltage-gated calcium channels (VGCCs). Similarly complex postsynaptic molecular machinery is also required. This includes the presence of ionotropic and metabotropic receptors that are capable of detecting and translating the presynaptic message (neurotransmitters) into various postsynaptic events, ranging from changes in resting membrane potential to gene expression, thus amplifying the presynaptic signal. b | Electrical transmission is mediated by clusters of intercellular channels called gap junctions that connect the interior of two adjacent cells, and thereby directly enable the bidirectional passage of electrical currents carried by ions (arrows) as well as intracellular messengers and small metabolites (not illustrated here). Electrical synapses are bidirectional in nature: when a presynaptic action potential propagates to the postsynaptic cell, the membrane resting potential of the postsynaptic cell simultaneously propagates to the presynaptic cell. Escape networks Neural networks found in invertebrate and vertebrate nervous systems (usually containing a small number of cells that include sensory and motor neurons) that seem optimized to mediate fast escape behaviours. Mauthner cell A large reticulospinal neuron found in teleost fish that mediates (among other functions) tail-flip sensory-evoked escape responses. occurs in the retina30 and the primary afferents of escape networks31–33. Gap junction-mediated lateral excitation has also been shown to occur in a network of cerebellar interneurons, promoting the spread of chemically mediated synaptic inputs between remote dendritic arborizations34. Because of their reliability (transmission at electrical synapses is not probabilistic in nature) and the absence of a synaptic delay (transmission occurs instantaneously), electrical synapses are also a typical feature of escape response networks in both invertebrates33,35,36 and vertebrates37. Together, these findings indicate that, from a functional point of view, no particular form of transmission is ‘better’ than the other one. Rather, the mechanisms of chemical and electrical transmission seem to be specifically adapted to communicate different aspects of cellular processing and functioning. Structural complexity of electrical synapses. Bolstered by the implicit molecular complexity of chemical transmission and its ability to undergo plasticity, there was until recently the widespread notion that electrical transmission is a rather simple, static and rigid form of neuronal communication. However, studies in cell expression systems38–43 and in neurons44 have shown that gap junctions are highly dynamic, complex structures and that their channels are actively turned over. More specifically, ultrastructural and functional analysis of mixed (electrical and chemical) synapses on the goldfish Mauthner cell (a large reticulospinal neuron in fish37,45) revealed that — as observed at gap junctions formed by recombinant CX43 in cell expression systems43 — gap junction hemichannels are simultaneously added at the edges of gap junction plaques (clusters of intercellular gap junction channels), where they dock with hemichannels in the apposed membrane to form cell–cell channels, and that intact junctional regions are removed from the centre of these plaques into either the presynaptic axon or the postsynaptic Mauthner cell dendrite44. Moreover, in this study, electrical coupling was readily modified by peptides that interfere with endocytosis or exocytosis, suggesting that the strength of electrical synapses at these terminals is determined, at least in part, by a fast turnover of gap junction channels44. Indeed, the half-life of this turnover was estimated to be 1–3 hours44, a time consistent with that observed for other ion channels and synaptic receptors46, indicating that this dynamic process could be involved in regulating electrical transmission (FIG. 2). Thus, as it occurs with postsynaptic receptors at glutamatergic chemical synapses for the regulation of their synaptic strength47–51, regulated trafficking of hemichannels and intercellular channels could underlie modifications of gap junctional conductance. Furthermore, like their chemical counterparts, electrical synapses are highly modifiable by the action of neuromodulators such as dopamine and are capable of activity-dependent plasticity (see REF. 52 for a review). 252 | APRIL 2014 | VOLUME 15 www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS Postsynaptic density (PSD). Originally named after its identification by electron microscopy, the term refers to a macromolecular complex that supports postsynaptic function at chemical synapses and includes neurotransmitters receptors, scaffolding proteins and regulatory signalling molecules. PSD95 (Postsynaptic density protein 95). A protein that contains multiple domains that mediate its association with receptors, cell-adhesion molecules and cytoplasmic signalling molecules. By virtue of these interactions, it influences the surface delivery, stability and subcellular location of postsynaptic receptors, and facilitates their functional coupling to downstream signalling pathways. The existence of such dynamic structural and functional properties indicates that electrical transmission must rely on more than the intercellular channels. Constitutive trafficking and regulated trafficking of ligand-gated ion channels to and from the plasma membrane are important processes for the maintenance of chemical synaptic function. At glutamatergic synapses, this requires interactions between the glutamate receptors’ carboxyl termini and various cytosolic proteins and scaffolding proteins50,53,54, which are mostly located at the postsynaptic density (PSD)55 (FIG. 2a). Similarly, electrical synapses require the molecular machinery to mediate the turnover of gap junction channels. Gap junctions are now considered to be a part of multiprotein complexes51–53. Electron-dense areas similar to those of PSDs, referred to as ‘semi-dense cytoplasmic matrices’ (REF. 56), have been observed at neuronal gap junctions when explored by electron microscopy (FIG. 2b). The detailed composition of these PSD-like structures is largely unknown57, although several molecules are known to interact with CX36 (REFS 58–61) and its teleost homologue62, most notably zonula occludens protein 1 (ZO1)59,60,62 and calcium/calmodulin-dependent protein kinase II (CaMKII)63,64. ZO1 is a scaffolding protein of the MAGUK family that is known to associate with many connexins65, and it might have a similar role in neuronal gap junctions to that of PSD95 at glutamatergic synapses. Conserved regions of the C terminus of CX36 (and its two teleost homologues)61,62,66 mediate its interactions with ZO1 and are required for the insertion of new gap junction proteins into electrical synapses44,66. ZO1 has also been proposed to have an essential role a Chemical synapse in regulating the transition from connexon to intercellular gap junction channel formation in the periphery of CX43‑containing junctions67. In contrast to the permanent structural role of ZO1, CaMKII has been proposed to be a non-obligatory component of electrical synapses, with its association being linked to synaptic activity (see below)64. Associated regulatory proteins seem to be crucial for innexin-based electrical transmission too, as a stomatin-like protein (UNC‑1) was shown to be required for proper channel function at gap junctions formed by UNC‑9 (an innexin) in Caenorhabditis elegans68,69. Recent data suggest that, similarly to pre- and postsynaptic sites at chemical synapses, one side of an electrical synapse should not necessarily be considered to be the mirror image of the other 70. Molecular asymmetries in neuronal gap junctions suggest that electrical synapses are functionally diverse. The intercellular channels at electrical synapses can in some cases be constructed by the apposition of two different connexins, forming ‘heterotypic’ (as opposed to ‘homotypic’) gap junctions14. Gap junctions at auditory mixed synapses of the goldfish Mauthner cell are constructed by the apposition of hemichannels formed by two homologues of mammalian CX36 (CX35 and CX34.7)70; whereas CX35 was restricted to presynaptic hemiplaques, CX34.7 was found only in postsynaptic hemiplaques, and together they formed heterotypic junctions70. Heterotypic gap junction channels have been associated with rectification of electrical transmission70–73, which is defined as the propensity of some electrical synapses to display differential resistance to b Electrical synapse Hemichannel insertion Glutamate Semi-dense cytoplasmic matrix Gap junction channel ZO1 AMPA receptor PSD Regulated trafficking PSD95 CaMKII Removal Figure 2 | Trafficking of channels at chemical and electrical synapses. a | Ionotropic glutamate receptors are trafficked Nature Reviews | Neuroscience in and out of synapses. Postsynaptic densities (PSDs) provide a scaffold that helps to regulate this trafficking. PSD95 and calcium/calmodulin-dependent protein kinase II (CaMKII) are both abundant components of PSDs. Regulated trafficking of AMPA receptors is thought to underlie the modification of synaptic strength at glutamatergic synapses. b | There is turnover of gap junction channels at electrical synapses. New connexons are trafficked as unpaired hemichannels to the membrane in vesicles, where they are inserted at the periphery of the gap junction plaque and dock with hemichannels in the apposing membrane. They are internalized as small clusters of entire channels (light grey) into either of the coupled cells from regions near the centre of the plaque. Various proteins make up the ‘semi-dense cytoplasmic matrix’, which acts as a scaffold for these channels. Zonula occludens protein 1 (ZO1) is a structural component of this scaffold. By contrast, CaMKII (not shown) seems to be a non-obligatory component of the macromolecular complex, with functions that might be similar to those at PSDs of chemical synapses. NATURE REVIEWS | NEUROSCIENCE VOLUME 15 | APRIL 2014 | 253 © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS current flow in one versus the other direction across the junction. Thus, such molecular asymmetry can support rectification of electrical transmission, which, by favouring the spread of membrane responses from the Mauthner cell to presynaptic endings, promotes cooperativity between auditory afferents. This association was initially observed in invertebrates 71 and is also supported by a wealth of data on heterotypic channels formed by recombinant connexins in cell expression systems72–74. Although asymmetry based on the presence of two CX36 homologues is restricted to teleost fish (teleost fish underwent an additional genome duplication75), the finding provided unambiguous evidence that vertebrate electrical synapses can be asymmetrical and that such asymmetry might underlie important functional properties. Molecular asymmetry in electrical synapses might not be restricted to the existence of molecularly different connexons but could also result from post-translational modifications of individual connexins or differences in the complement of gap junction-associated proteins forming the semi-dense cytoplasmic matrix57, endowing electrical transmission with more complex properties. In summary, although chemical synapses seem to be structurally more complex and functionally dynamic, emerging evidence indicates that electrical synapses might be similarly complex, diverse and highly modifiable. Electroretinography An extracellularly recorded electrical response that reflects the activation of various cells in the retina (including photoreceptors, inner retinal cells and the output ganglion cells) in response to visual stimulation. Synaptic interactions during development Gap junctional communication between neurons is developmentally regulated: it is initially prominent but declines at later developmental stages76,77. In the rodent brain, this form of intercellular communication is widespread between postnatal days 5 and 12 and dramatically decreases after postnatal day 16 to remain restricted to some cell types76,78–81. The initial increase in coupling enables developing neurons to form functional domains that exhibit coordinated patterns of spontaneous activity 79,82,83. It has been proposed that the formation of these transient domains and networks might serve as a developmental blueprint, influencing various developmental cellular phenomena, including cell contact inhibition, neuronal differentiation, migration, circuit formation and the elimination of chemical synapses77. The formation of transient gap junction networks is a common feature of invertebrate84–87 and vertebrate76,77,80,81 nervous systems. In C. elegans, a transient network formed by the innexin gap junction protein NSY‑5 (also known as innexin‑19 (INX‑19)) is required for the left and right olfactory AWC neurons to create asymmetrical patterns of gene expression during embryogenesis87. NSY‑5 mutants failed to establish this asymmetry, suggesting that intercellular communication mediated by NSY‑5 junctions is a critical developmental requirement 87. Similarly, transient gap junctions mediate avoidance of segmental homologous cell projections during development in the leech88. Upon contact, the processes of AP neurons, two identifiable homologous motor neuron-like cells, stop growing and finally retract, thus defining the topography of their neuronal arbors89–91. Recent data suggest that transient inx1‑containing gap junctions that are formed between the processes of homologous AP cells mediate this developmental phenomenon88. Neurite growth arrest did not occur in animals in which inx1 was knocked down by the expression of a doublestrand interfering RNA (dsRNA) or in those expressing an inx1 mutant that lacked open channel pore function but retained the adhesive cellular binding properties of gap junction channels88. These findings suggest that these gap junctions act as conduits for an unknown messenger molecule that mediates avoidance of segmental homologous cell projections. Another well-established example of a transient gap junction network can be found in the developing mammalian spinal cord92–94. Developing lumbar motor neurons communicate by gap junctions, leading to synchronous patterns of nerve activity during a period in which muscle fibres are innervated by axons originating from multiple motor neurons92,94. The transition from multiple innervation to the characteristic single innervation in muscle fibres occurs at late embryonic and early postnatal stages and is an activity-dependent process, which results from competition between active synapses95. It has been shown that a transient gap junction network modulates this developmental transition from multiple- to single-fibre innervation96. Gap junctional coupling between motor neurons disappears at late embryonic and early postnatal stages, causing a lack of correlated activity between innervating axons (elimination cannot occur if synapses remain synchronously activated), which in turn paves the way for synapse competition and subsequent synapse elimination in multiply innervated muscle fibres96. Reduction of gap junctional communication with gap junction-blocking agents disrupted synchronous patterns of nerve activity and enhanced synapse elimination96. Consistent with this finding, acceleration of synapse elimination was observed in mice lacking expression of CX40 (REF. 97), a connexin that is expressed in developing motor neurons and the expression of which dramatically decreases after birth92. Together, these findings indicate that transient gap junction networks have a primary role in gating the transition from multiple- to single-fibre muscle innervation. Remarkably, the formation and subsequent elimination of gap junction coupling between neurons was found to be correlated with the emergence of chemical neurotransmission in several species. This robust phenomenon was again observed in developing invertebrate 84,98,99 and vertebrate 100–102 nervous systems. Furthermore, several studies indicate that gap junction proteins expressed during development are required for the formation of chemical synapses. Flies containing mutations in the innexin genes shaking B and ogre failed to establish appropriate functional synaptic connections in the visual system, as shown by electroretinography103. This developmental defect could be rescued by transgenic expression of the lost innexins during pulpal development but not at later developmental stages103. More-direct evidence of the relationship between electrical synapses and the formation of 254 | APRIL 2014 | VOLUME 15 www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS chemical synapses comes from motor neurons in the snail Helisoma trivolvis98. Manipulations that led to a reduction of coupling between these snail motor neurons accelerated the formation of chemical synapses, whereas exposure to cholinergic antagonists (which block chemical neurotransmission at these synapses) caused prolonged maintenance of electrical transmission98, suggesting an inverse relationship between the establishment of chemical and electrical connectivity at early stages of synaptic development. This relationship between the formation of electrical and chemical synapses is also observed in vertebrates. The presence of gap junction coupling between developing spinal motor neurons also followed an inverse relationship with the formation of chemical synapses100. The decline in motor neuron coupling that occurs at late embryonic and early postnatal stages is correlated with the formation of glutamatergic synapses in these spinal circuits100. Furthermore, exposure to NMDA receptor antagonists arrested the developmental decline in gap junction coupling between spinal motor neurons, suggesting that the increase in glutamatergic synaptic activity that is generally associated with the onset of locomotion promotes the disappearance of gap junction-mediated networks between developing motor neurons100. The exquisite relationship between the early presence of electrical connections and the subsequent formation of chemical synapses has recently been demonstrated in the leech. The sequential development of electrical and chemical synapses between two types of identifiable neurons that participate in a behaviourally relevant local-bending circuit offered the possibility of directly addressing this issue84. Synaptic transmission between the P cell (that transduces pressure) and the motor neuron-like AP cell is initially exclusively electrical but becomes predominantly chemical in the adult animal. Although electrical transmission remains in adulthood, it is considerably weakened99. Silencing inx1 in embryonic P cells when innexin-based gap junctions are initially forming and chemical synapses are absent (at ‘~50%’ of the way through embryonic development, a stage defined by external morphological features after egg deposition) prevented not only the establishment of electrical synapses but also the development of chemical transmission99 (FIG. 3a), elegantly demonstrating the requirement of electrical synapses for the formation of chemical contacts. The development of networks mediated by chemical synapses in the mouse olfactory bulb was arrested in CX36‑knockout mice101, in which electrical transmission is greatly reduced. This indicates that the early presence of electrical synapses is also a prerequisite for the formation of chemical synapses in mammals. This phenomenon was also observed in the developing neocortex. Consecutive divisions of individual radial glial progenitor cells normally produce sister excitatory neurons, which leads to the formation of ontogenetic cortical columns that preferentially develop specific chemical synapses with each other rather than with nearby non-siblings102. Sister excitatory neurons are initially electrically coupled102, and blockade of this communication impairs the subsequent formation of specific chemical synapses between sister excitatory neurons in ontogenetic cortical columns102. Finally, hippocampal long-term potentiation of glutamatergic transmission was shown to be impaired in CX36‑knockout mice, suggesting that functional properties of chemical synapses are also affected by the absence of electrical synapses at early developmental stages104. I have discussed the mechanistic relationship between the presence of electrical synapses and the subsequent formation of chemical connections. However, conversely, receptors for chemical neurotransmitters and chemical synapses have been shown to regulate the emergence and disappearance of gap junctions during development105–107. In vitro studies of cortical and hypothalamic neurons indicate that, despite the fact that chemical connections are not yet formed, activation of specific receptors for chemical neurotransmitters regulates the expression of CX36 at early developmental stages106. Prolonged (2 week)106 activation of group II metabotropic glutamate receptors (mGluRs) leads to an increase in the expression of CX36 through a cAMP–protein kinase A (PKA)-dependent intracellular pathway, whereas activation of type A GABA receptors (GABAARs) leads to a reduction of CX36 expression106 (FIG. 3b), indicating that the formation of gap junctional networks relies on the interplay between these two types of receptors. Because GABAARs are depolarizing at these early developmental stages, their effects are mediated via calcium influx and activation of PKC106. This developmental regulation of Cx36 expression involves regulation of both gene transcription (mGluR-dependent increase) and protein translation (GABAAR-dependent decrease)106. These changes were proposed to be specific for these receptors, and other classes of glutamate, GABA and acetylcholine receptors are presumed not to be involved106. Activation of glutamate receptors also reduces coupling observed at late developmental stages. As mentioned above, blocking NMDA receptors delayed the formation of chemical synapses in spinal motor neurons100, suggesting that the emergence of glutamatergic transmission regulates uncoupling. This is supported by findings in the rat hypothalamus105, in which activation of NMDA receptors at late developmental stages led to cAMP-responsive element-binding protein (CREB)-dependent downregulation of Cx36 expression105, providing a direct mechanistic relationship between the emergence of chemical transmission and the disappearance of neuronal gap junction coupling (FIG. 3b). In summary, the development of neural circuits in disparate nervous systems seems to rely critically on interactions between chemical and electrical synapses, which reciprocally and dynamically regulate the emergence of these two forms of transmission (FIG. 3c). Synaptic interactions in the adult nervous system The effects of neuromodulators on gap junction communication provide a clear example of how chemical and electrical transmission interact in the adult nervous system. Neuromodulators are released from presynaptic terminals and activate specific metabotropic G proteincoupled receptors, which trigger signalling cascades NATURE REVIEWS | NEUROSCIENCE VOLUME 15 | APRIL 2014 | 255 © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS a b 100 ms Uninjected Sham Scramble RNAi c Group II mGluR High 6 mV AP Amount of CX36 GJC or level of channel activity P cell Low GABAAR P1 NMDAR P15 P30 Time Electrical synapse Chemical synapse Gap junctions provide a blueprint for the formation of neuronal networks Neurotransmitter 1 Ca2+ Synaptic vesicle Gap junction channel Ionotropic receptor 2 Metabotropic receptor The formation of chemical synapses triggers the elimination of coupling in most neuronal cell types Figure 3 | Electrical and chemical synapses interact during development. a | Blockade of electrical synapse formation Nature Reviews | Neuroscience in leech embryos perturbs the formation of chemical synapses. Chemical synaptic potentials in an AP cell in response to a single spike in a P cell (marked by the grey bar on the AP recordings) under control conditions (uninjected, sham-injected and injected with scrambled double-stranded RNA) and after injection of double-stranded RNA (RNAi) that interferes with the translation of innexin 1 in embryonic P cells, at a developmental time (~50% of the way through embryonic development, a stage defined by external morphological features after egg deposition) at which innexin-based gap junctions are forming but chemical synapses have not yet formed. b | Effects of signalling through type A GABA receptors (GABAARs), metabotropic glutamate receptors (mGluRs) and NMDA receptors (NMDARs) on gap junction communication (GJC) during development. The red line represents the increase (upward phase) and decrease (downward phase) in the amount of neuronal GJC and expression of connexin 36 (CX36) during postnatal development. The arrows show the direction of the change in GJC after activation of the given receptor. P1, P15 and P30 indicate postnatal days 1, 15 and 30. c | Forms of interactions between electrical and chemical synapses during development. Gap junctions lead first (step 1) to the formation of neural circuits. The arrival of chemical synapses (step 2) leads to the elimination of gap junctions in most neuronal types. Part a is reproduced, with permission, from REF. 99 © (2010) Society for Neuroscience. Part b is modified, with permission, from REF. 107 © (2013) Elsevier. that modulate synaptic communication between target cells (FIG. 4a). Dopamine, for example, has been shown to modulate gap junction communication in various vertebrate and invertebrate cell types16,19 (see REFS 16,102 for reviews), suggesting that both connexin and innexin gap junction proteins are targets of its regulatory actions. At electrical synapses mediated by CX36 or CX57, and their teleost homologues, activation of dopamine can lead to either an increase or a decrease in gap junctional conductance between neurons through a cAMPdependent mechanism that involves the activation of PKA. That is, dopamine was observed to enhance coupling at goldfish electrical synapses108,109 and to promote decoupling between retinal horizontal cells110–112. Two phosphorylation sites in CX36, Ser110 and Ser293 (corresponding to Ser110 and Ser276, respectively, in its fish homologues112,113), seem to be essential for the effects of dopamine on electrical synapses. Recent evidence indicates that dopamine, acting via dopamine D1 and D5 receptors, can either promote PKA-mediated phosphorylation of CX36 at regulatory sites, leading to an increase in coupling, or promote PKA-induced activation of protein phosphatase 2A and subsequent dephosphorylation of CX36, leading to decoupling 114. This finding therefore provides a mechanism for dopamine-induced bidirectional regulation of the strength of electrical synapses. Dopamine can also activate D2 receptors, which leads to a reduction in cAMP and PKA activation, and increases coupling between rods and cones in the vertebrate retina115. Other neuromodulatory systems that regulate the strength of electrical synapses include noradrenaline116, serotonin117,118, histamine119 and nitric oxide120,121. 256 | APRIL 2014 | VOLUME 15 www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS a Chemical synapse b Electrical synapse Synaptic varicosity Mixed synapse Neuromodulator Glutamate Synaptic vesicle Gap junction channel NMDAR Neurotransmitter Ionotropic receptor Postsynaptic density Metabotropic receptor cc d Heterosynaptic interaction CaMKII Goldfish mixed synapse Dopaminergic varicosity Dopamine NMDAR or mGluR-mediated signalling mGluR CB1R eCB release D1R or D5R PKA Figure 4 | Types of interactions between electrical and chemical synapses in the adult nervous Nature Reviewssystem. | Neuroscience a | Neurotransmitter modulators released by nearby synaptic terminals (purple) regulate the synaptic strength of chemical and electrical synapses through activation of G protein-coupled metabotropic receptors. Regulation at chemical synapses could occur pre- or postsynaptically. b | Electrical and chemical synapses coexist at mixed synapses. Glutamatergic synapses regulate the strength of electrical synapses by a postsynaptic mechanism that includes the activation of NMDA receptors (NMDARs) and calcium/calmodulin-dependent protein kinase II (CaMKII). c | Regulation of electrical synapses by glutamatergic transmission could also be heterosynaptic. Nearby glutamatergic synapses can regulate electrical transmission through NMDAR or metabotropic glutamate receptor (mGluR) activation. d | Another mechanism of interaction at goldfish mixed synapses results when synaptic activity leads to mGluR activation, which in turn triggers endocannabinoid (eCB) release from the postsynaptic cell, and activates cannabinoid type 1 receptors (CB1Rs) on nearby dopaminergic fibres. CB1R activation leads to dopamine release that, by activating postsynaptic dopamine D1 receptors (D1Rs) and D5Rs and increasing protein kinase A (PKA) activity, is responsible for simultaneous enhancement of electrical and glutamatergic synaptic transmission. ON bipolar cells Retinal cells that functionally link photoreceptors (cones and rods) to ganglion cells. ON bipolar cells are excited by the release of glutamate from photoreceptors, whereas OFF bipolar cells are instead inhibited. All type amacrine cells The AII is a type of amacrine cell (a class of retinal interneuron) that relays rod-driven information through the ON‑centre cone bipolar axons to ON‑centre ganglion cells (output neurons of the retina) via electrical synapses. Regulation of electrical coupling by neuromodulators can have profound functional consequences. Chemical and electrical synapses coexist in various circuits, raising the question of how these two networks interact to generate function122. In the rabbit retina, rod and cone inputs were shown to converge on cone bipolar cells19. In the case of the cones, the connection is direct via ON bipolar cells, whereas for the rods, the connection is indirect. This indirect connection specifically relies on electrical synapses between AII type amacrine cells themselves and between AII type amacrine cells and ON bipolar cells123. Both homologous and heterologous synapses are regulated by dopamine (gap junctions between AII type amacrine cells) and nitric oxide (AII type amacrine cells-to-ON bipolar gap junctions), respectively, efficiently reconfiguring retinal circuits by switching between direct and indirect pathways123,124. Remarkably, it has been shown that fast, amino acidmediated chemical transmission also interacts with electrical synapses. ‘Mixed’ chemical and electrical synapses found at ‘large myelinated club endings’ (or simply ‘club endings’)52 — terminations of primary auditory afferents on the teleost Mauthner cells — provide a unique opportunity to study interactions between these two modalities of synaptic transmission52,125 (FIG. 4b). Activation of these synapses with high-frequency bursts (500 Hz) of activity leads to long-lasting potentiation of both the electrical and glutamatergic components of the mixed synaptic potential evoked by the activation of these terminals126,127. Thus, both forms of transmission at these contacts exhibit activity-dependent plasticity. Strikingly, potentiation of both electrical and chemical transmission was blocked by NMDA receptor antagonists126,127, suggesting that potentiation of electrical transmission is initiated by the activity of the coexisting glutamatergic NATURE REVIEWS | NEUROSCIENCE VOLUME 15 | APRIL 2014 | 257 © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS synapses. The potentiations also required an increase in postsynaptic calcium levels and the activation of CaMKII128 (FIG. 4b). CaMKII activation was shown to lead to an increase in gap junction conductance not only at goldfish club endings but also in CX36‑containing gap junctions in cell expression systems129. Consistent with these findings, CaMKII was found to phosphorylate CX36 at retinal electrical synapses at sites that enhance coupling between these cells130. CaMKII is an abundant component of the PSD at chemical synapses and has been implicated in mechanisms of activity-dependent plasticity at chemical synapses55,131. In similarity to its interaction with the NR2B (also known as GluN2B) subunit of the NMDA receptor at PSDs, CaMKII interacts with CX36 by binding to its cytoplasmic domains63,131. There are multiple CaMKIIα phosphorylation sites in CX36 and its fish homologues CX35 and CX34.7 (REFS 63,64). Ser315 in CX36, Ser298 in CX35 and Ser300 in CX34.7 are exclusive phosphorylation sites for CaMKII (that is, they are not targeted by other kinases)63,64. By contrast, Ser110 and Ser293 in CX36, Ser110 and Ser276 in CX35, and Ser110 and Ser277 in CX34.7 can be phosphorylated by CaMKII and PKA63,64. Mixed synapses provided unambiguous evidence that glutamatergic and electrical synapses interact. Because interactions occur postsynaptically (FIG. 4b), it is possible that similar interactions could also take place hetero synaptically between neighbouring glutamatergic and electrical synapses (FIG. 4c). Supporting this possibility, ultrastructural studies have revealed that the proximity of NR1‑containing PSDs to gap junctions in the rat inferior olive and retina is comparable to the distances between such PSDs and gap junctions in goldfish club endings132,133. Indeed, NMDA receptor–CaMKII-dependent modulation of gap junction coupling has been proposed to take place in the inferior olive132,134 and has been shown more recently to occur in the retina130; that is, presynaptic activity of glutamatergic ON bipolar cells increases phosphorylation of CX36 in AII type amacrine cells130. Glutamatergic synapses have also been shown to induce an increase in dye coupling between hypothalamic neurons135. Moreover, glutamatergic transmission has been shown to promote activity-dependent long-term depression of electrical coupling, indicating that interactions between glutamatergic and electrical synapses are diverse and widespread. That is, high-frequency activation of corticothalamic inputs triggered long-term depression of electrical transmission between thalamic relay neurons, which required the activation of mGluRs136. Interestingly, the interactions between chemical and electrical synapses were club ending-specific and did not spread to neighbouring club endings137, suggesting that the mechanisms of PSD-initiated interactions with gap junctions are short-ranged and take place within less than 5 μm, the usual distance between two neighbouring club endings in the smooth Mauthner cell dendrite. Given that plastic changes triggered by glutamate are terminal-specific and club ending afferents can be differentially activated by sounds, the gap junctions at these contacts coexist at different degrees of conductance on the Mauthner cell dendrite137. These findings suggest that a single cell can be differentially coupled to its neighbours based on the activity of chemical synapses located in the proximity of each gap junction. Inferior olivary cells, which connect to up to 40 other inferior olivary cells, have been shown to be differentially coupled with their partners. Such heterogeneity in coupling strength has also been reported in the rabbit retina114 and the rat cerebellar cortex 138. In summary, by regulating nearby gap junctions, PSDs at glutamatergic synapses provide a mechanism for fine-tuning electrical coupling within networks of electrically coupled neurons. This form of regulation, by determining the probability with which neurons interact with each other, can reconfigure networks and create functional compartments of neurons that have a propensity to cooperate. Heterosynaptic interactions between electrical and chemical synapses can be very complex and involve intricate interactions between glutamatergic synapses and neurotransmitter modulators. High-frequency stimulation of club endings potentiates both electrical and chemical transmission. This effect is dependent on the activation of mGluR1 receptors139 and the formation of endocannabinoids, which, by acting on cannabinoid type 1 receptors (CB1Rs), lead to the release of dopamine from varicosities located in the close vicinity of the club endings and Mauthner cell dendrites139. The release of dopamine from nearby varicosities in turn leads to the potentiation of the electrical (and glutamatergic) synaptic response through a cAMP–PKA-mediated postsynaptic mechanism139 (FIG. 4d). This mechanism of potentiation illustrates the complexity and possibilities of interactions between chemical and electrical synapses. It also demonstrates that the local availability of dopamine is not exclusively under the control of dopaminergic neurons but that it is also determined by local synaptic activity (both electrical and chemical). Finally, the ‘shunting’ effect of inhibitory synapses provides another example of interactions between chemical and electrical synapses140–142. Chemical inhibitory synaptic conductance often short-circuits the currents that are generated at adjacent excitatory synapses by locally increasing membrane conductance140,142. Depending on the anatomical arrangement, inhibitory conductances can dramatically reduce coupling if they are in the proximity of electrical synapses by ‘shunting’ electrical currents before they have had the chance to spread to a nearby neuron through the gap junctions (FIG. 5) (see REF. 16 for a review). This form of interaction was shown to occur within the inferior olive glomerulus140,141 and between expansion motor neurons of the mollusc Navanax inermis142, in which quick and transient decoupling sculpts clusters of functionally related neurons involved in phasic behaviours. Further emphasizing the functional significance of these fast interactions, reciprocal electrical and inhibitory synapses organize networks of mammalian GABAergic interneurons143. Modelling one of these interneuron networks in the cerebellum suggested that combining fast inhibitory synapses and electrical coupling can promote synchronized gamma oscillations143, which have been associated with cognitive processing and are affected in some pathological conditions144. 258 | APRIL 2014 | VOLUME 15 www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS a GABAergic terminal GABA GABAAR Gap junction channel b Reduction in electrical coupling ↑gCl– Current shunting Figure 5 | Interactions between electrical synapses and inhibitory chemical synapses. a | Inhibitory GABAergic Nature Reviews | Neuroscience synapses are often located in the vicinity of gap junctions between dendrodendritic processes (spines) of two neurons. b | By locally increasing membrane conductance, inhibitory synaptic chloride conductances (gCl−) produced by activation of type A GABA receptors (GABAARs) briefly shunt excitatory currents to temporarily reduce effective electrical coupling between two coupled neurons. Associative binding The term refers to tasks of episodic memory that require the associative combining of distinct components into a compound episode. Are interactions involved in dysfunction? The strong developmental inter-relationship between electrical and chemical synapses suggests that its disruption might underlie neurological and psychiatric conditions that are acquired during development. Accordingly, lack or dysfunction of electrical transmission could lead, as shown in the neocortex 102 and CX36‑knockout mice101, to subtle deficits in the formation of mature neural circuits. Interestingly, CX36 has been associated with juvenile myoclonic epilepsy, a generalized form of epilepsy with onset in early adolescence145,146, but the exact mechanistic linkage remains unknown. It has been suggested that some single-nucleotide polymorphisms may influence CX36 expression and contribute to the pathogenesis of this disease by affecting the normal formation of neural circuits145. In addition, dysfunction of glutamatergic transmission, which triggers the developmental decline in gap junctional communication via NMDA receptor activation105, could result in abnormal levels of electrical coupling in some brain regions that might lead to increased neuronal synchrony and seizures147. As previously mentioned, CX36‑mediated electrical synapses are a feature of inhibitory interneurons in the adult mammalian brain, including the neocortex, thalamus and hippocampus6–8,136. Inhibitory interneurons are crucial for various brain processes, as they contribute to the generation of gamma-frequency oscillatory activity (30–80 Hz), which has been associated with cognitive processing 144,148. High-frequency network oscillatory synchronizations seem to be crucial for defining the conscious state149 and for associative binding in learning and memory. Strikingly, these cortical gamma oscillations are impaired in CX36‑knockout mice150, suggesting a role for electrical synapses in generating this rhythm. Alteration of oscillatory activity has been proposed to contribute to the pathophysiology of schizophrenia148,151, Parkinson’s disease152 and autism spectrum disorders153. Moreover, dysfunction of glutamatergic154–156 and dopaminergic157,158 transmission is involved in these conditions, suggesting that dysregulation of electrical coupling could contribute to the underlying pathophysiological processes. The investigation of these possibilities could lead to the identification of novel therapeutic opportunities for treating these conditions. Recent data suggest that the interactions between chemical and electrical synapses that are observed during development have an important functional role after brain insult. Strikingly, networks of electrically coupled neurons identified during development can reappear after brain injury. Adult cat spinal motor neurons become recoupled by gap junctions after peripheral nerve injury 159, re-establishing the transient gap junction network observed during development. Gap junction coupling was proposed to have a beneficial functional role by helping to maintain the viability of axotomized motor neurons until synaptic connections with their muscle fibre targets are re-established159. Although it is unclear if this axotomy-induced spinal network disappears after nerve regeneration, glutamatergic transmission was shown to increase expression of CX36 and gap junction communication in in vivo and in vitro models of ischaemic stroke and traumatic brain injury 107. The increase in CX36 expression occurs about 2 hours after injury and is likely to be triggered by the massive release of glutamate from injured neurons107. As observed during development 106, activation of group II mGluRs has also been shown to mediate this increase in gap junction communication160. These interactions between glutamatergic and electrical transmission are thought to control death and survival mechanisms in injured neurons160. A wealth of data indicate that activation of NMDA receptors mediates the glutamate toxicity that occurs in the injured brain161. Interestingly, electrical synapses seem to be involved in this phenomenon. The death of forebrain neurons evoked by intraperitoneal injection of NMDA was greatly attenuated by gap junction blockers and in CX36‑knockout mice162,163, suggesting that gap junctional coupling could have a deleterious effect on neuronal survival. It has been suggested that neurodegenerative signals could be passed to other, NATURE REVIEWS | NEUROSCIENCE VOLUME 15 | APRIL 2014 | 259 © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS a b Neuronal injury Glutamate release NMDAR activation Electrical synapse Group II mGluR activation Lack or dysfunction of gap junction channels might lead to defective formation of neural circuits and underlie developmentally related neurological conditions Chemical synapse Ca2+ Ca2+ overload CX36 expression Gap junction-permeable ‘death signals’ Penumbra neuron Gap junction channel Neuronal gap junction coupling Ionotropic receptor Enhanced neuronal damage Damaged neuron Gap junction Synaptic vesicle Neurotransmitter Metabotropic receptor Glutamate-induced enhancement of coupling afer injury or stroke leads to neuronal death Dysregulation of electrical synapses by chemical synapses might contribute to cognitive dysfunction Figure 6 | Interactions between chemical and electrical synapses and pathological processes. a | Chemical and electrical synapses interact after injury. Release of glutamate from injured neurons causes neuronal damage via NMDA Nature Reviews | Neuroscience receptor (NMDAR) activation and calcium overload at the site of the lesion, and the generation of gap junction-permeable ‘death signals’. Simultaneous enhancement of neuronal gap junction coupling, via metabotropic glutamate receptor (mGluR)-dependent increased expression of connexin 36 (CX36), extends neuronal damage by facilitating the passage of these ‘death signals’ to neurons adjacent to the lesion or in the ‘penumbra’. b | A summary of potential interactions between chemical and electrical synapses during pathological processes. The lack, or dysfunction, of gap junction channels at early developmental stages might lead to defective formation of critical neural circuits formed by chemical synapses, underlying developmentally related neurological conditions. During adulthood, release of glutamate from compromised neurons after injury or stroke enhances coupling, which leads to neuronal death in uncompromised areas adjacent to the lesion. Dysregulation of electrical synapses’ strength by neurotransmitters might contribute to cognitive disorders. Part a is modified, with permission, from REF. 107 © (2013) Elsevier. undamaged neurons (neurons adjacent to the lesion or ‘penumbra’ area) as a result of the increase in neuronal coupling triggered by the activation of group II mGluRs164 (FIG. 6a). Conclusions Electrical synapses have proved to be more widespread in the mammalian brain than originally anticipated. Thus, networks of electrically and chemically coupled neurons are not restricted to invertebrates but seem to be a feature of all nervous systems. This realization emphasizes the need to further explore the functional properties of electrical synapses (of which we know significantly less than we do about chemical synapses) and their interactions with chemical synapses. Studies of mixed synapses provide unambiguous evidence that, rather than unconnected lines of communication, electrical and chemical synapses cooperate and extensively interact, and that the speed and reliability of electrical transmission can be combined with the ability of chemical transmission to induce plastic changes. Mixed synapses are widespread in cold-blooded vertebrates52 but they seem to be less numerous in mammals, perhaps because of the much-improved speed of transmission observed at mammalian chemical synapses165. However, there is growing evidence of their presence in the mammalian nervous system, including in the spinal cord166, in a subset of hippocampal mossy fibres167,168 and, interestingly, between dendrites of hippocampal inhibitory interneurons169. Interactions between electrical and chemical synapses are complex and diverse, and occur at all stages of brain development. During development, interactions between electrical synapses are crucial for the formation of neural circuits, whereas such interactions in the adult brain result in dynamic reconfiguration of hardwired networks. These interactions are likely to have important pathological implications (FIG. 6b). Recapitulation of some developmental interactions between chemical and electrical synapses after brain injury indicates that, rather than being exceptional, these interactions constitute basic and necessary mechanisms of communication in the nervous system. Furthermore, the interactions between dopaminergic, glutamatergic and electrical synapses that have been observed in the adult brain highlight their importance in reconfiguring neural circuits, and their dysregulation could contribute to cognitive impairment. Future studies are likely to confirm the intimate relationship between the chemical and electrical modalities of synaptic transmission and shed further light on their contribution to disease states. 260 | APRIL 2014 | VOLUME 15 www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS Sheng, M., Sabatini, B. L. & Sudhof, T. C. (eds) The Synapse (Cold Spring Harbor Laboratory, 2012). 2. Bennett, M. V. L. & Zukin, R. S. Electrical coupling and neuronal synchronization in the mammalian brain. Neuron 41, 495–511 (2004). 3.Zoli, M. et al. The emergence of the volume transmission concept. Brain Res. Brain Res. Rev. 26, 136–147 (1998). 4. Faber, D. S. & Korn, H. Electrical field effects: their relevance in central neural networks. Physiol. Rev. 69, 821–863 (1989). 5. Connors, B. W. & Long, M. A. Electrical synapses in the mammalian brain. Annu. Rev. Neurosci. 27, 393–418 (2004). 6. Galarreta, M. & Hestrin, S. A network of fast-spiking cells in the neocortex connected by electrical synapses. Nature 402, 72–75 (1999). 7. Gibson, J. R., Beierlein, M. & Connors, B. W. Two networks of electrically coupled inhibitory neurons in neocortex. Nature 402, 75–79 (1999). 8. Galarreta, M. & Hestrin, S. Electrical synapses between GABA-releasing interneurons. Nature Rev. Neurosci. 2, 425–433 (2001). 9. Bennett, M. V. Electrical synapses, a personal perspective (or history). Brain Res. Brain Res. Rev. 32, 16–28 (2000). 10. Bandara, H. M. H. N., Lam, O. L. T., Jin, L. J. & Samaranayake, L. Microbial chemical signaling: a current perspective. Crit. Rev. Microbiol. 38, 217–249 (2012). 11. Li, Z. & Nair, S. K. Quorum sensing: how bacteria can coordinate activity and synchronize their response to external signals? Protein Sci. 21, 1403–1417 (2012). 12. Dustin, M. L. Signaling at neuro/immune synapses. J. Clin. Invest. 122, 1149–1155 (2012). 13. Sterling, P. & Matthews, G. Structure and function of ribbon synapses. Trends Neurosci. 28, 20–29 (2005). 14. Goodenough, D. A. & Paul, D. L. Gap junctions. Cold Spring Harb. Perspect. Biol. 1, a002576 (2009). 15. MacVicar, B. A. & Thompson, R. J. Non-junction functions of pannexin‑1 channels. Trends Neurosci. 33, 93–102 (2010). 16.Pereda, A. E. et al. Gap junction-mediated electrical transmission: regulatory mechanisms and plasticity. Biochim. Biophys. Acta 1828, 134–146 (2013). 17. Shimizu, K. & Stopfer, M. Gap junctions. Curr. Biol. 23, R1026–R1031 (2013). 18. Söhl, G., Maxeiner, S. & Willecke, K. Expression and functions of neuronal gap junctions. Nature Rev. Neurosci. 6, 191–200 (2005). 19. Bloomfield, S. A. & Völgyi, B. The diverse functional roles and regulation of neuronal gap junctions in the retina. Nature Rev. Neurosci. 10, 495–506 (2009). 20. Condorelli, D. F., Belluardo, N., Trovato-Salinaro, A. & Mudò, G. Expression of Cx36 in mammalian neurons. Brain Res. Brain Res. Rev. 32, 72–85 (2000). 21. Lee, S.‑C., Cruikshank, S. J. & Connors, B. W. Electrical and chemical synapses between relay neurons in developing thalamus. J. Physiol. 588, 2403–2415 (2010). 22. Curti, S., Hoge, G., Nagy, J. I. & Pereda, A. E. Synergy between electrical coupling and membrane properties promotes strong synchronization of neurons of the mesencephalic trigeminal nucleus. J. Neurosci. 32, 4341–4359 (2012). 23. Phelan, P. Innexins: members of an evolutionarily conserved family of gap-junction proteins. Biochim. Biophys. Acta 1711, 225–245 (2005). 24.Kandarian, B. et al. The medicinal leech genome encodes 21 innexin genes: different combinations are expressed by identified central neurons. Dev. Genes Evol. 222, 29–44 (2012). 25.Liu, P. et al. Six innexins contribute to electrical coupling of C. elegans body-wall muscle. PLoS ONE 8, e76877 (2013). 26. Getting, P. A. Modification of neuron properties by electrotonic synapses. I. Input resistance, time constant, and integration. J. Neurophysiol. 37, 846–857 (1974). 27. Getting, P. A. & Willows, A. O. Modification of neuron properties by electrotonic synapses. II. Burst formation by electrotonic synapses. J. Neurophysiol. 37, 858–868 (1974). 28. Galarreta, M. & Hestrin, S. Spike transmission and synchrony detection in networks of GABAergic interneurons. Science 292, 2295–2299 (2001). The authors propose that electrical synapses operate as coincidence detectors in networks of electrically coupled neurons. 1. 29. Veruki, M. L. & Hartveit, E. All (rod) amacrine cells form a network of electrically coupled interneurons in the mammalian retina. Neuron 33, 935–946 (2002). 30. DeVries, S. H., Qi, X., Smith, R., Makous, W. & Sterling, P. Electrical coupling between mammalian cones. Curr. Biol. 12, 1900–1907 (2002). 31. Pereda, A. E., Bell, T. D. & Faber, D. S. Retrograde synaptic communication via gap junctions coupling auditory afferents to the Mauthner cell. J. Neurosci. 15, 5943–5955 (1995). 32. Curti, S. & Pereda, A. E. Voltage-dependent enhancement of electrical coupling by a subthreshold sodium current. J. Neurosci. 24, 3999–4010 (2004). 33. Herberholz, J., Antonsen, B. L. & Edwards, D. H. A lateral excitatory network in the escape circuit of crayfish. J. Neurosci. 22, 9078–9085 (2002). 34. Vervaeke, K., Lorincz, A., Nusser, Z. & Silver, R. A. Gap junctions compensate for sublinear dendritic integration in an inhibitory network. Science 335, 1624–1628 (2012). 35.Phelan, P. et al. Mutations in shaking‑B prevent electrical synapse formation in the Drosophila giant fiber system. J. Neurosci. 16, 1101–1113 (1996). 36. Edwards, D. H., Heitler, W. J. & Krasne, F. B. Fifty years of a command neuron: the neurobiology of escape behavior in the crayfish. Trends Neurosci. 22, 153–161 (1999). 37. Faber, D. S. & Pereda, A. E. in Encyclopedia of Fish Physiology: From Genome Environment (ed. Farrell, A.) 73–79 (Elsevier, 2011). 38. Laird, D. W. Life cycle of connexins in health and disease. Biochem. J. 394, 527–543 (2006). 39. Laird, D. W. The life cycle of a connexin: gap junction formation, removal, and degradation. J. Bioenerg. Biomembr. 28, 311–318 (1996). 40. Gumpert, A. M., Varco, J. S., Baker, S. M., Piehl, M. & Falk, M. M. Double-membrane gap junction internalization requires the clathrin-mediated endocytic machinery. FEBS Lett. 582, 2887–2892 (2008). 41.Piehl, M. et al. Internalization of large doublemembrane intercellular vesicles by a clathrindependent endocytic process. Mol. Biol. Cell 18, 337–347 (2007). 42.Lauf, U. et al. Dynamic trafficking and delivery of connexons to the plasma membrane and accretion to gap junctions in living cells. Proc. Natl Acad. Sci. USA 99, 10446–10451 (2002). 43.Gaietta, G. et al. Multicolor and electron microscopic imaging of connexin trafficking. Science 296, 503–507 (2002). 44.Flores, C. E. et al. Trafficking of gap junction channels at a vertebrate electrical synapse in vivo. Proc. Natl Acad. Sci. USA 109, E573–E582 (2012). This paper provides the first evidence of the trafficking of gap junction channels in a native electrical synapse. 45. Pereda, A. E. & Faber, D. S. in Encyclopedia of Fish Physiology: From Genome Environment (ed. Farrell, A.) 66–72 (Elsevier, 2011). 46. Hervé, J.‑C., Derangeon, M., Bahbouhi, B., Mesnil, M. & Sarrouilhe, D. The connexin turnover, an important modulating factor of the level of cell‑to‑cell junctional communication: comparison with other integral membrane proteins. J. Membr. Biol. 217, 21–33 (2007). 47.Lüscher, C. et al. Role of AMPA receptor cycling in synaptic transmission and plasticity. Neuron 24, 649–658 (1999). This article provides one of the first descriptions of the existence of trafficking and cycling of receptors at glutamatergic synapses and their potential roles in synaptic plasticity. 48. Carroll, R. C. & Zukin, R. S. NMDA-receptor trafficking and targeting: implications for synaptic transmission and plasticity. Trends Neurosci. 25, 571–577 (2002). 49. Carroll, R. C., Beattie, E. C., von Zastrow, M. & Malenka, R. C. Role of AMPA receptor endocytosis in synaptic plasticity. Nature Rev. Neurosci. 2, 315–324 (2001). 50.Chen, L. et al. Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms. Nature 408, 936–943 (2000). 51. Ehlers, M. D. Reinsertion or degradation of AMPA receptors determined by activity-dependent endocytic sorting. Neuron 28, 511–525 (2000). 52. Pereda, A. E., Rash, J. E., Nagy, J. I. & Bennett, M. V. L. Dynamics of electrical transmission at club endings on the Mauthner cells. Brain Res. Brain Res. Rev. 47, 227–244 (2004). NATURE REVIEWS | NEUROSCIENCE 53.Dong, H. et al. GRIP: a synaptic PDZ domaincontaining protein that interacts with AMPA receptors. Nature 386, 279–284 (1997). 54. Leonard, A. S., Davare, M. A., Horne, M. C., Garner, C. C. & Hell, J. W. SAP97 is associated with the α-amino‑3‑hydroxy-5‑methylisoxazole-4‑propionic acid receptor GluR1 subunit. J. Biol. Chem. 273, 19518–19524 (1998). 55. Kennedy, M. B. The postsynaptic density at glutamatergic synapses. Trends Neurosci. 20, 264–268 (1997). 56. Sotelo, C. & Korn, H. Morphological correlates of electrical and other interactions through low-resistance pathways between neurons of the vertebrate central nervous system. Int. Rev. Cytol. 55, 67–107 (1978). 57. Lynn, B. D., Li, X. & Nagy, J. I. Under construction: building the macromolecular superstructure and signaling components of an electrical synapse. J. Membr. Biol. 245, 303–317 (2012). The authors review the proteins associated with connexin-formed gap junction channels at electrical synapses. 58. Li, X., Lynn, B. D. & Nagy, J. I. The effector and scaffolding proteins AF6 and MUPP1 interact with connexin36 and localize at gap junctions that form electrical synapses in rodent brain. Eur. J. Neurosci. 35, 166–181 (2012). 59. Li, X., Lu, S. & Nagy, J. I. Direct association of connexin36 with zonula occludens‑2 and zonula occludens‑3. Neurochem. Int. 54, 393–402 (2009). 60. Li, X., Olson, C., Lu, S. & Nagy, J. I. Association of connexin36 with zonula occludens‑1 in HeLa cells, βTC‑3 cells, pancreas, and adrenal gland. Histochem. Cell Biol. 122, 485–498 (2004). 61.Li, X. et al. Neuronal connexin36 association with zonula occludens‑1 protein (ZO‑1) in mouse brain and interaction with the first PDZ domain of ZO‑1. Eur. J. Neurosci. 19, 2132–2146 (2004). 62. Flores, C. E., Li, X., Bennett, M. V. L., Nagy, J. I. & Pereda, A. E. Interaction between connexin35 and zonula occludens‑1 and its potential role in the regulation of electrical synapses. Proc. Natl Acad. Sci. USA 105, 12545–12550 (2008). 63.Alev, C. et al. The neuronal connexin36 interacts with and is phosphorylated by CaMKII in a way similar to CaMKII interaction with glutamate receptors. Proc. Natl Acad. Sci. USA 105, 20964–20969 (2008). The paper provides evidence for the existence of direct protein–protein interactions between gap junction-forming proteins and CaMKII, a regulatory kinase that also regulates chemical synapses. 64.Flores, C. E. et al. Variability of distribution of Ca2+/ calmodulin-dependent kinase II at mixed synapses on the mauthner cell: colocalization and association with connexin 35. J. Neurosci. 30, 9488–9499 (2010). 65. Hervé, J.‑C., Bourmeyster, N. & Sarrouilhe, D. Diversity in protein-protein interactions of connexins: emerging roles. Biochim. Biophys. Acta 1662, 22–41 (2004). 66.Helbig, I. et al. In vivo evidence for the involvement of the carboxy terminal domain in assembling connexin 36 at the electrical synapse. Mol. Cell. Neurosci. 45, 47–58 (2010). 67. Rhett, J. M., Jourdan, J. & Gourdie, R. G. Connexin 43 connexon to gap junction transition is regulated by zonula occludens‑1. Mol. Biol. Cell 22, 1516–1528 (2011). 68. Chen, B., Liu, Q., Ge, Q., Xie, J. & Wang, Z.‑W. UNC‑1 regulates gap junctions important to locomotion in C. elegans. Curr. Biol. 17, 1334–1339 (2007). 69. Norman, K. R. & Maricq, A. V. Innexin function: minding the gap junction. Curr. Biol. 17, R812–R814 (2007). 70.Rash, J. E. et al. Molecular and functional asymmetry at a vertebrate electrical synapse. Neuron 79, 957–969 (2013). The paper provides evidence suggesting that gap junction hemiplaques at electrical synapses might not necessarily be the mirror image of each other. 71.Phelan, P. et al. Molecular mechanism of rectification at identified electrical synapses in the Drosophila giant fiber system. Curr. Biol. 18, 1955–1960 (2008). 72.Barrio, L. C. et al. Gap junctions formed by connexins 26 and 32 alone and in combination are differently affected by applied voltage. Proc. Natl Acad. Sci. USA 88, 8410–8414 (1991). 73. Oh, S., Rubin, J. B., Bennett, M. V., Verselis, V. K. & Bargiello, T. A. Molecular determinants of electrical rectification of single channel conductance in gap junctions formed by connexins 26 and 32. J. Gen. Physiol. 114, 339–364 (1999). VOLUME 15 | APRIL 2014 | 261 © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS 74. Verselis, V. K., Ginter, C. S. & Bargiello, T. A. Opposite voltage gating polarities of two closely related connexins. Nature 368, 348–351 (1994). 75. Volff, J.‑N. Genome evolution and biodiversity in teleost fish. Heredity (Edinb.) 94, 280–294 (2005). 76. Kandler, K. & Katz, L. C. Neuronal coupling and uncoupling in the developing nervous system. Curr. Opin. Neurobiol. 5, 98–105 (1995). 77. Montoro, R. J. & Yuste, R. Gap junctions in developing neocortex: a review. Brain Res. Brain Res. Rev. 47, 216–226 (2004). 78. Peinado, A., Yuste, R. & Katz, L. C. Extensive dye coupling between rat neocortical neurons during the period of circuit formation. Neuron 10, 103–114 (1993). The paper describes the existence of extensive gap junction coupling and its developmental regulation in the vertebrate brain. 79. Peinado, A., Yuste, R. & Katz, L. C. Gap junctional communication and the development of local circuits in neocortex. Cereb. Cortex 3, 488–498 (1993). 80. Penn, A. A., Wong, R. O. & Shatz, C. J. Neuronal coupling in the developing mammalian retina. J. Neurosci. 14, 3805–3815 (1994). 81. Bittman, K., Owens, D. F., Kriegstein, A. R. & LoTurco, J. J. Cell coupling and uncoupling in the ventricular zone of developing neocortex. J. Neurosci. 17, 7037–7044 (1997). 82. Yuste, R., Peinado, A. & Katz, L. C. Neuronal domains in developing neocortex. Science 257, 665–669 (1992). 83. Yuste, R. & Katz, L. C. Control of postsynaptic Ca2+ influx in developing neocortex by excitatory and inhibitory neurotransmitters. Neuron 6, 333–344 (1991). 84. Marin-Burgin, A., Eisenhart, F. J., Baca, S. M., Kristan, W. B. & French, K. A. Sequential development of electrical and chemical synaptic connections generates a specific behavioral circuit in the leech. J. Neurosci. 25, 2478–2489 (2005). 85. Marin-Burgin, A., Eisenhart, F. J., Kristan, W. B. & French, K. A. Embryonic electrical connections appear to pre-figure a behavioral circuit in the leech CNS. J. Comp. Physiol. A Neuroethol. Sens. Neural Behav. Physiol. 192, 123–133 (2006). 86. Wolszon, L. Cell–cell interactions define the innervation patterns of central leech neurons during development. J. Neurobiol. 27, 335–352 (1995). 87. Chuang, C.‑F., Vanhoven, M. K., Fetter, R. D., Verselis, V. K. & Bargmann, C. I. An innexindependent cell network establishes left–right neuronal asymmetry in C. elegans. Cell 129, 787–799 (2007). 88. Baker, M. W., Yazdani, N. & Macagno, E. R. Gap junction-dependent homolog avoidance in the developing CNS. J. Neurosci. 33, 16673–16683 (2013). 89. Wolszon, L. R., Rehder, V., Kater, S. B. & Macagno, E. R. Calcium wave fronts that cross gap junctions may signal neuronal death during development. J. Neurosci. 14, 3437–3448 (1994). 90. Wolszon, L. R., Gao, W. Q., Passani, M. B. & Macagno, E. R. Growth cone “collapse” in vivo: are inhibitory interactions mediated by gap junctions? J. Neurosci. 14, 999–1010 (1994). 91. Wolszon, L. R., Passani, M. B. & Macagno, E. R. Interactions during a critical period inhibit bilateral projections in embryonic neurons. J. Neurosci. 15, 1506–1515 (1995). 92. Chang, Q., Gonzalez, M., Pinter, M. J. & Balice-Gordon, R. J. Gap junctional coupling and patterns of connexin expression among neonatal at lumbar spinal motor neurons. J. Neurosci. 19, 10813–10828 (1999). 93. Personius, K., Chang, Q., Bittman, K., Panzer, J. & Balice-Gordon, R. Gap junctional communication among motor and other neurons shapes patterns of neural activity and synaptic connectivity during development. Cell Commun. Adhes. 8, 329–333 (2001). 94. Walton, K. D. & Navarrete, R. Postnatal changes in motoneurone electrotonic coupling studied in the in vitro rat lumbar spinal cord. J. Physiol. 433, 283–305 (1991). 95. Colman, H. & Lichtman, J. W. Interactions between nerve and muscle: synapse elimination at the developing neuromuscular junction. Dev. Biol. 156, 1–10 (1993). 96. Personius, K. E. & Balice-Gordon, R. J. Loss of correlated motor neuron activity during synaptic competition at developing neuromuscular synapses. Neuron 31, 395–408 (2001). 97. Personius, K. E., Chang, Q., Mentis, G. Z., O’Donovan, M. J. & Balice-Gordon, R. J. Reduced gap junctional coupling leads to uncorrelated motor neuron firing and precocious neuromuscular synapse elimination. Proc. Natl Acad. Sci. USA 104, 11808–11813 (2007). 98. Szabo, T. M., Faber, D. S. & Zoran, M. J. Transient electrical coupling delays the onset of chemical neurotransmission at developing synapses. J. Neurosci. 24, 112–120 (2004). Using a reduced invertebrate model, the authors unambiguously demonstrate the inter-relationship between the formation of electrical and chemical synapses. 99. Todd, K. L., Kristan, W. B. Jr & French, K. A. Gap junction expression is required for normal chemical synapse formation. J. Neurosci. 30, 15277–15285 (2010). This study elegantly demonstrates the initial requirement of electrical synapses for the formation of chemical synapses in an in vivo system. 100.Mentis, G. Z., Díaz, E., Moran, L. B. & Navarrete, R. Increased incidence of gap junctional coupling between spinal motoneurones following transient blockade of NMDA receptors in neonatal rats. J. Physiol. 544, 757–764 (2002). The paper provides evidence for the existence of an inverse relationship between the presence of electrical synapses and chemical synapses in the mammalian brain. 101. Maher, B. J., McGinley, M. J. & Westbrook, G. L. Experience-dependent maturation of the glomerular microcircuit. Proc. Natl Acad. Sci. USA 106, 16865–16870 (2009). The paper demonstrates the existence of deficits in the formation of circuits formed by chemical synapses in mice lacking the gap junction protein CX36. 102.Yu, Y.‑C. et al. Preferential electrical coupling regulates neocortical lineage-dependent microcircuit assembly. Nature 486, 113–117 (2012). 103.Curtin, K. D., Zhang, Z. & Wyman, R. J. Gap junction proteins expressed during development are required for adult neural function in the Drosophila optic lamina. J. Neurosci. 22, 7088–7096 (2002). 104.Wang, Y. & Belousov, A. B. Deletion of neuronal gap junction protein connexin 36 impairs hippocampal LTP. Neurosci. Lett. 502, 30–32 (2011). 105.Arumugam, H., Liu, X., Colombo, P. J., Corriveau, R. A. & Belousov, A. B. NMDA receptors regulate developmental gap junction uncoupling via CREB signaling. Nature Neurosci. 8, 1720–1726 (2005). The paper describes at the mechanistic level how the emergence of glutamatergic transmission leads to a massive reduction in gap junction coupling in the developing brain. 106.Park, W.‑M. et al. Interplay of chemical neurotransmitters regulates developmental increase in electrical synapses. J. Neurosci. 31, 5909–5920 (2011). 107. Belousov, A. B. & Fontes, J. D. Neuronal gap junctions: making and breaking connections during development and injury. Trends Neurosci. 36, 227–236 (2013). 108.Pereda, A., Triller, A., Korn, H. & Faber, D. S. Dopamine enhances both electrotonic coupling and chemical excitatory postsynaptic potentials at mixed synapses. Proc. Natl Acad. Sci. USA 89, 12088–12092 (1992). 109.Pereda, A. E., Nairn, A. C., Wolszon, L. R. & Faber, D. S. Postsynaptic modulation of synaptic efficacy at mixed synapses on the Mauthner cell. J. Neurosci. 14, 3704–3712 (1994). 110. Piccolino, M., Neyton, J. & Gerschenfeld, H. M. Decrease of gap junction permeability induced by dopamine and cyclic adenosine 3’:5’-monophosphate in horizontal cells of turtle retina. J. Neurosci. 4, 2477–2488 (1984). 111. Lasater, E. M. & Dowling, J. E. Dopamine decreases conductance of the electrical junctions between cultured retinal horizontal cells. Proc. Natl Acad. Sci. USA 82, 3025–3029 (1985). 112.Urschel, S. et al. Protein kinase A‑mediated phosphorylation of connexin36 in mouse retina results in decreased gap junctional communication between AII amacrine cells. J. Biol. Chem. 281, 33163–33171 (2006). 113. Kothmann, W. W., Li, X., Burr, G. S. & O’Brien, J. Connexin 35/36 is phosphorylated at regulatory sites in the retina. Vis. Neurosci. 24, 363–375 (2007). 114. Kothmann, W. W., Massey, S. C. & O’Brien, J. Dopamine-stimulated dephosphorylation of connexin 262 | APRIL 2014 | VOLUME 15 36 mediates AII amacrine cell uncoupling. J. Neurosci. 29, 14903–14911 (2009). 115. Ribelayga, C., Cao, Y. & Mangel, S. C. The circadian clock in the retina controls rod-cone coupling. Neuron 59, 790–801 (2008). 116. Zsiros, V. & Maccaferri, G. Noradrenergic modulation of electrical coupling in GABAergic networks of the hippocampus. J. Neurosci. 28, 1804–1815 (2008). 117. Rörig, B. & Sutor, B. Serotonin regulates gap junction coupling in the developing rat somatosensory cortex. Eur. J. Neurosci. 8, 1685–1695 (1996). 118. Johnson, B. R., Peck, J. H. & Harris-Warrick, R. M. Amine modulation of electrical coupling in the pyloric network of the lobster stomatogastric ganglion. J. Comp. Physiol. A 172, 715–732 (1993). 119. Hatton, G. I. & Yang, Q. Z. Synaptically released histamine increases dye coupling among vasopressinergic neurons of the supraoptic nucleus: mediation by H1 receptors and cyclic nucleotides. J. Neurosci. 16, 123–129 (1996). 120.O’Donnell, P. & Grace, A. A. Cortical afferents modulate striatal gap junction permeability via nitric oxide. Neuroscience 76, 1–5 (1997). 121.Rörig, B. & Sutor, B. Nitric oxide-stimulated increase in intracellular cGMP modulates gap junction coupling in rat neocortex. Neuroreport 7, 569–572 (1996). 122.Bargmann, C. I. Beyond the connectome: how neuromodulators shape neural circuits. Bioessays 34, 458–465 (2012). 123.Xia, X.‑B. & Mills, S. L. Gap junctional regulatory mechanisms in the AII amacrine cell of the rabbit retina. Vis. Neurosci. 21, 791–805 (2004). 124.Mills, S. L. & Massey, S. C. Differential properties of two gap junctional pathways made by AII amacrine cells. Nature 377, 734–737 (1995). 125.Pereda, A. et al. Connexin35 mediates electrical transmission at mixed synapses on Mauthner cells. J. Neurosci. 23, 7489–7503 (2003). 126.Yang, X. D., Korn, H. & Faber, D. S. Long-term potentiation of electrotonic coupling at mixed synapses. Nature 348, 542–545 (1990). This study provides the first evidence for the existence of activity-dependent potentiation in electrical synapses. 127.Pereda, A. E. & Faber, D. S. Activity-dependent shortterm enhancement of intercellular coupling. J. Neurosci. 16, 983–992 (1996). 128.Pereda, A. E. et al. Ca2+/calmodulin-dependent kinase II mediates simultaneous enhancement of gapjunctional conductance and glutamatergic transmission. Proc. Natl Acad. Sci. USA 95, 13272–13277 (1998). The paper provides the first evidence for the role of CaMKII in regulating electrical transmission. 129.Del Corsso, C., Iglesias, R., Zoidl, G., Dermietzel, R. & Spray, D. C. Calmodulin dependent protein kinase increases conductance at gap junctions formed by the neuronal gap junction protein connexin36. Brain Res. 1487, 69–77 (2012). 130.Kothmann, W. W. et al. Nonsynaptic NMDA receptors mediate activity-dependent plasticity of gap junctional coupling in the AII amacrine cell network. J. Neurosci. 32, 6747–6759 (2012). 131.Lisman, J., Yasuda, R. & Raghavachari, S. Mechanisms of CaMKII action in long-term potentiation. Nature Rev. Neurosci. 13, 169–182 (2012). 132.Hoge, G. J. et al. The extent and strength of electrical coupling between inferior olivary neurons is heterogeneous. J. Neurophysiol. 105, 1089–1101 (2011). 133.Rash, J. E. et al. High-resolution proteomic mapping in the vertebrate central nervous system: close proximity of connexin35 to NMDA glutamate receptor clusters and co‑localization of connexin36 with immunoreactivity for zonula occludens protein‑1 (ZO‑1). J. Neurocytol. 33, 131–151 (2004). 134.Urbano, F. J., Leznik, E. & Llinás, R. R. Modafinil enhances thalamocortical activity by increasing neuronal electrotonic coupling. Proc. Natl Acad. Sci. USA 104, 12554–12559 (2007). 135.Hatton, G. I. & Yang, Q. Z. Activation of excitatory amino acid inputs to supraoptic neurons. I. Induced increases in dye-coupling in lactating, but not virgin or male rats. Brain Res. 513, 264–269 (1990). 136.Landisman, C. E. & Connors, B. W. Long-term modulation of electrical synapses in the mammalian thalamus. Science 310, 1809–1813 (2005). 137.Smith, M. & Pereda, A. E. Chemical synaptic activity modulates nearby electrical synapses. Proc. Natl Acad. Sci. USA 100, 4849–4854 (2003). www.nature.com/reviews/neuro © 2014 Macmillan Publishers Limited. All rights reserved REVIEWS 138.Vervaeke, K. et al. Rapid desynchronization of an electrically coupled interneuron network with sparse excitatory synaptic input. Neuron 67, 435–451 (2010). 139.Cachope, R., Mackie, K., Triller, A., O’Brien, J. & Pereda, A. E. Potentiation of electrical and chemical synaptic transmission mediated by endocannabinoids. Neuron 56, 1034–1047 (2007). The article describes interactions between glutamatergic, dopaminergic and electrical synapses, demonstrating a complex functional inter-relationship between chemical and electrical transmission. 140.Llinas, R., Baker, R. & Sotelo, C. Electrotonic coupling between neurons in cat inferior olive. J. Neurophysiol. 37, 560–571 (1974). 141.Best, A. R. & Regehr, W. G. Inhibitory regulation of electrically coupled neurons in the inferior olive is mediated by asynchronous release of GABA. Neuron 62, 555–565 (2009). 142.Spira, M. E., Spray, D. C. & Bennett, M. V. Electrotonic coupling: effective sign reversal by inhibitory neurons. Science 194, 1065–1067 (1976). 143.Bartos, M. et al. Fast synaptic inhibition promotes synchronized gamma oscillations in hippocampal interneuron networks. Proc. Natl Acad. Sci. USA 99, 13222–13227 (2002). 144.Buzsáki, G. & Wang, X.‑J. Mechanisms of gamma oscillations. Annu. Rev. Neurosci. 35, 203–225 (2012). 145.Mas, C. et al. Association of the connexin36 gene with juvenile myoclonic epilepsy. J. Med. Genet. 41, e93 (2004). This study provides the first evidence of the relationship between CX36 and juvenile myoclonic epilepsy. 146.Hempelmann, A., Heils, A. & Sander, T. Confirmatory evidence for an association of the connexin‑36 gene with juvenile myoclonic epilepsy. Epilepsy Res. 71, 223–228 (2006). 147.Dudek, F. E., Snow, R. W. & Taylor, C. P. Role of electrical interactions in synchronization of epileptiform bursts. Adv. Neurol. 44, 593–617 (1986). 148.Nakazawa, K. et al. GABAergic interneuron origin of schizophrenia pathophysiology. Neuropharmacology 62, 1574–1583 (2012). 149.Steriade, M. Synchronized activities of coupled oscillators in the cerebral cortex and thalamus at different levels of vigilance. Cereb. Cortex 7, 583–604 (1997). 150.Hormuzdi, S. G. et al. Impaired electrical signaling disrupts gamma frequency oscillations in connexin 36‑deficient mice. Neuron 31, 487–495 (2001). The study provides evidence for the contribution of electrical coupling between cortical inhibitory interneurons in the generation of gamma oscillations, which are associated with cognitive phenomena. 151.Gonzalez-Burgos, G., Hashimoto, T. & Lewis, D. A. Alterations of cortical GABA neurons and network oscillations in schizophrenia. Curr. Psychiatry Rep. 12, 335–344 (2010). 152.Hammond, C., Bergman, H. & Brown, P. Pathological synchronization in Parkinson’s disease: networks, models and treatments. Trends Neurosci. 30, 357–364 (2007). 153.Welsh, J. P., Ahn, E. S. & Placantonakis, D. G. Is autism due to brain desynchronization? Int. J. Dev. Neurosci. 23, 253–263 (2005). 154.Moghaddam, B. Bringing order to the glutamate chaos in schizophrenia. Neuron 40, 881–884 (2003). 155.Javitt, D. C. et al. Translating glutamate: from pathophysiology to treatment. Sci. Transl. Med. 3, 102mr2 (2011). 156.Moghaddam, B. & Javitt, D. From revolution to evolution: the glutamate hypothesis of schizophrenia and its implication for treatment. Neuropsychopharmacology 37, 4–15 (2012). 157.Nieoullon, A. Dopamine and the regulation of cognition and attention. Prog. Neurobiol. 67, 53–83 (2002). 158.Penzes, P., Buonanno, A., Passafaro, M., Sala, C. & Sweet, R. A. Developmental vulnerability of synapses and circuits associated with neuropsychiatric disorders. J. Neurochem. 126, 165–182 (2013). 159.Chang, Q., Pereda, A., Pinter, M. J. & Balice-Gordon, R. J. Nerve injury induces gap junctional coupling among axotomized adult motor neurons. J. Neurosci. 20, 674–684 (2000). 160.Wang, Y. et al. Neuronal gap junction coupling is regulated by glutamate and plays critical role in cell death during neuronal injury. J. Neurosci. 32, 713–725 (2012). The authors describe the relationship between glutamate and increased gap junctional coupling observed after neuronal injury and its underlying mechanisms. 161.Hazell, A. S. Excitotoxic mechanisms in stroke: an update of concepts and treatment strategies. Neurochem. Int. 50, 941–953 (2007). 162.Belousov, A. B. et al. Neuronal gap junctions play a role in the secondary neuronal death following controlled cortical impact. Neurosci. Lett. 524, 16–19 (2012). 163.Wang, Y. et al. Neuronal gap junctions are required for NMDA receptor-mediated excitotoxicity: implications in ischemic stroke. J. Neurophysiol. 104, 3551–3556 (2010). 164.Belousov, A. B. Novel model for the mechanisms of glutamate-dependent excitotoxicity: role of neuronal gap junctions. Brain Res. 1487, 123–130 (2012). NATURE REVIEWS | NEUROSCIENCE 165.Sabatini, B. L. & Regehr, W. G. Timing of neurotransmission at fast synapses in the mammalian brain. Nature 384, 170–172 (1996). 166.Rash, J. E. et al. Mixed synapses discovered and mapped throughout mammalian spinal cord. Proc. Natl Acad. Sci. USA 93, 4235–4239 (1996). 167.Hamzei-Sichani, F. et al. Mixed electrical–chemical synapses in adult rat hippocampus are primarily glutamatergic and coupled by connexin‑36. Front. Neuroanat. 6, 13 (2012). 168.Vivar, C., Traub, R. D. & Gutiérrez, R. Mixed electrical– chemical transmission between hippocampal mossy fibers and pyramidal cells. Eur. J. Neurosci. 35, 76–82 (2012). 169.Fukuda, T. & Kosaka, T. Gap junctions linking the dendritic network of GABAergic interneurons in the hippocampus. J. Neurosci. 20, 1519–1528 (2000). 170.Valenstein, E. S. The War of the Soups and the Sparks: The Discovery of Neurotransmitters and the Dispute Over How Nerves Communicate (Columbia Univ. Press, 2006). 171.Elliott, T. R. The action of adrenalin. J. Physiol. 32, 401–467 (1905). 172.Loewi, O. Über humorale Übertragbarkeit der Herznervenwirkung. Pflugers Arch. Gesamte Physiol. Menschen Tiere 204, 629–640 (in German) (1924). 173.Sherrington, C. S. The Integrative Action of the Nervous System — Primary Source Edition (Nabu, 2013). 174.Katz, B. The Release of Neural Transmitter Substances (Sherrington Lecture) (Liverpool Univ. Press, 1969). 175.Furshpan, E. J. & Potter, D. D. Transmission at the giant motor synapses of the crayfish. J. Physiol. 145, 289–325 (1959). 176.Bennett, M. V., Aljure, E., Nakajima, Y. & Pappas, G. D. Electrotonic junctions between teleost spinal neurons: electrophysiology and ultrastructure. Science 141, 262–264 (1963). 177.Robertson, J. D., Bodenheimer, T. S. & Stage, D. E. The ultrastructure of Mauthner cell synapses and nodes in goldfish brains. J. Cell Biol. 19, 159–199 (1963). 178.Furshpan, E. J. “Electrical transmission” at an excitatory synapse in a vertebrate brain. Science 144, 878–880 (1964). Acknowledgements This research was supported by US National Institutes of Health grants DC03186, DC011099, NS055726, NS085772 and NS0552827 to A.E.P. Competing interests statement The author declares no competing interests. VOLUME 15 | APRIL 2014 | 263 © 2014 Macmillan Publishers Limited. All rights reserved