Download In vitro and in vivo Microelectrode Array Recording Techniques

In vitro and in vivo Microelectrode
Array Recording Techniques
43rd Annual SfN Meeting
Satellite Event
Monday, November 11, 2013
6:30 pm - 8:30 pm
San Diego Convention Center
Room: 24A
Program
Use of Multi-Electrode Arrays for Large-Scale Genetic Studies of
Synaptic Transmission Phenotypes in Hippocampal Slices
Maksym Kopanitsa, PhD
Synome Ltd, Babraham Research Campus, Cambridge, UK
Optogenetic Stimulation: From Single Neurons to
Populations on MEAs
Thomas DeMarse, PhD
Department of Biomedical Engineering, University of Florida, USA
Simultaneous Stimulation and Detection of Axonal Action
Potential Propagation Using High-Density CMOS-Based
Microelectrode Arrays (‘Neurochip’)
Günther Zeck, PhD
Natural and Medical Sciences Institute, Reutlingen, Germany
Investigating the Physiology and Pathophysiology of the Ventral
Tegmental Area in Behaving Rodents
Stanislav Koulchitsky, PhD
University of Liège, Laboratory of Pharmacology, Liège, Belgium
Use of Multi-Electrode Arrays for Large-Scale
Genetic Studies of Synaptic Transmission
Phenotypes in Hippocampal Slices
Maksym Kopanitsa, PhD
Synome Ltd, Babraham Research Campus, Cambridge, UK
The importance of the genetic regulation of the nervous system has been widely
recognised. Due to evolutionary closeness to humans, mice are one of the most
favoured model organisms in genetics. Nonetheless, despite several decades of
electrophysiological experiments on hundreds of spontaneous and engineered
mouse mutants, the literature does not permit comparison of synaptic phenotypes
and thus no general principles describing genetic regulation of vertebrate synaptic
transmission have been formulated. Among reasons for this shortcoming are the
variability between laboratories in their electrophysiological methods and the
absence of large-scale studies comparing many mutations in the identical setting.
We conducted a large-scale study of synaptic transmission utilising CA3-CA1
fEPSPs recorded from hippocampal slices of genetically altered mice and we
have accumulated a standardised set of such measurements in over 50 such
mutants. In this presentation, I will show how problems of scale and variability
can be tackled by the use of multi-electrode array technology. Using examples
of phenotypes caused by genetic mutations, I will also consider how traditional
electrophysiological measurements, such as fEPSP size, paired-pulse facilitation
and long-term potentiation can be utilised for uniform quantitative comparisons
of multiple experimental conditions (e.g., comparing different mutations or drug
treatments).
Supported by the European Union Seventh Framework Programme under
grant agreement numbers HEALTH-F2-2009-241498 (“EUROSPIN” project),
HEALTH-F2–2009-242167 (“SynSys project”), and HEALTH-F2-2009-241995
(“GENCODYS project”).
References:
Kopanitsa MV, Afinowi NO, Grant SG. 2006, Recording long-term potentiation of synaptic transmission
by three-dimensional multi-electrode arrays. BMC Neurosci., 7:61.
Ryan TJ, Kopanitsa MV, Indersmitten T et al., 2013, Evolution of GluN2A/B cytoplasmic domains
diversified vertebrate synaptic plasticity and behavior. Nat Neurosci.16(1):25-32.
Optogenetic Stimulation:
From Single Neurons to
Populations on MEAs
Thomas DeMarse, PhD
Department of Biomedical Engineering, University of Florida, USA
Optogenetic tools are light-sensitive proteins derived from single-cell organisms.
Upon expression in mammalian neurons, they can be used to effectively control
neuronal excitability down to the millisecond timescale. There are two main classes
of optogenetic tools that are used to control depolarization and hyperpolarization,
and respectively generate or inhibit action potentials in selected populations of
neurons. By applying these tools within the MEA platform, it is now possible to
gain precise control over the excitation and inhibition of neural activity across large
neural populations down to nearly simultaneous and independent control over
individual neurons within that population.
This presentation will describe ongoing research in my laboratory using optogenetic
stimulation in dissociated culture and cortical/hippocampal slices for modulating
excitability using ChR2 and eNpHr. I will also cover specific strategies an MEA
researcher might use to facilitate the application of optogenetic tools within their
own research including transfection in vitro and in vivo, hardware for stimulation,
and various tips on implementing this new stimulation paradigm.
References:
Tye, K. M., & Deisseroth, K. (2012). Optogenetic investigation of neural circuits underlying brain
disease in animal models. Nature Reviews. Neuroscience, 13(4), 251-66. doi:10.1038/nrn3171
Simultaneous Stimulation and Detection of
Axonal Action Potential Propagation Using HighDensity CMOS-Based MEAs (‘Neurochip’)
Günther Zeck, PhD
Natural and Medical Sciences Institute, Reutlingen, Germany
CMOS based high density microelectrode arrays (MEAs) provide the opportunity to
electrically stimulate neurons (Eickenscheidt et al. 2012) and to record the occurrence and
propagation of action potentials in the stimulated neurons (Zeck et al. 2011). However,
simultaneous electrical stimulation and recording of many neurons has not been possible
so far.
Here we present stimulation and recording results obtained with a newly developed
electrode array which comprises 4225 recording sites (pitch 16 µm) interlaced with 1024
capacitive stimulation sites. The entire array is insulated by a thin, inert and biocompatible
oxide layer. Continuous recording of all sensors over several minutes at a sampling rate of
25 kHz is demonstrated. Electrical stimulation can be performed using arbitrary stimulus
shapes presented at different positions. The electrode area can be varied during the
experiment.
We selected the ex vivo guinea pig retina as an appropriate nervous tissue. Light-stimuli
applied to the retina elicit reliable action potentials in the ganglion cells, which are detected
by the multiple sensors on the array. These stimuli allow to precisely locating the position
of the ganglion cells. The axonal path is revealed in electrical images calculated from spike
triggered averages.
Based on the cell and axon position we demonstrate how stimuli of different polarity, of
different shape and position evoke action potentials in nearby or in distant neurons. Action
potential detection is achieved within a few hundred microseconds after stimulus onset.
The presented arrays enable bidirectional ‘closed-loop’- type interfacing with other
neuronal preparations to target specific sets of neurons.
References:
Zeck G, Lambacher A, Fromherz P. 2011, Axonal Transmission in the Retina Introduces a Small Dispersion of
Relative Timing in the Ganglion Cell Population Response. PLoS ONE 6(6): e20810.
Eickenscheidt M, Jenkner M, Thewes R, Fromherz P, Zeck G. 2012, Electrical stimulation of retinal neurons in
epiretinal and subretinal configuration using a multicapacitor array. J Neurophysiol 107: 2747-2755, 2012.
Investigating the Physiology and
Pathophysiology of the Ventral Tegmental Area
in Behaving Rodents
Stanislav Koulchitsky, PhD
University of Liège, Laboratory of Pharmacology, Liège, Belgium
The mesocorticolimbic reward system is known to mediate the reinforcement of
some behaviors which are directly related to the survival (search for food, foraging,
sexual activity, etc.). It was also shown to mediate an appreciation of some abstract
items lacking natural reward value, but allowing us to engage in the world, such as
music and other arts, self-development, etc. This system originates in the ventral
tegmental area (VTA) and comprises both dopaminergic (DA) and GABAergic
neurons.
Cocaine as well as the other addictive drugs „hijack“ this system, totally modifying
the behavior of the addicted person. Previous studies have shown that all drugs of
abuse increase the concentration of dopamine in the nucleus accumbens, a major
target area of the VTA, but the precise dynamics of VTA neurons and of the “VTA
network” during drug self-administration is unknown.
We are currently studying both neuronal activity in the VTA and some behavioral
parameters (i) during natural conditions in an open field and (ii) during acute and
chronic exposure to cocaine in awake, freely moving rats using the Multichannel
Wireless system. Both extracellular single unit activity and local field potentials are
recorded. The latter mostly consist of theta (~8 Hz) rhythms, interspersed with
gamma (~50 Hz) activity. In particular, coupling between the phase of the theta
rhythm and spiking of individual neurons is investigated. We have also started to
study the activity of the VTA during cocaine self-administration. During the talk,
some results on these various topics will be presented.
One of our next aims is to simultaneously record neuronal activity from the VTA
and major afferent brain areas, constituting the reward system. This will hopefully
allow us to decipher the spatio-temporal dynamics of this system.
The expected outcomes of our work are to: (i) reveal some mechanisms of cocaine
addiction and cocaine-induced behavior; (ii) better understand the functional
wiring of the reward system.
Notes
Speakers
Maksym Kopanitsa, PhD
Maksym Kopanitsa was born and educated in Ukraine. He received his PhD in Biophysics at the
Bogomoletz Institute of Physiology (Kyiv, Ukraine) in the laboratory of Prof. Oleg Krishtal where
he carried out research on pharmacology of voltage- and ligand-gated ion channels and synaptic
transmission. During his post-doctoral appointments in the labs of Prof. Jeremy Lambert (University of Dundee, 2003-2004) and Prof. Seth Grant (The Wellcome Trust Sanger Institute, 20042010) Maksym worked with mutant mice to study genetic regulation of the synaptic transmission
in brain slices. Now Maksym Kopanitsa works at Synome Ltd in Cambridge, UK. He leads a multidisciplinary team of scientists deciphering phenotypic effects of mutations and pharmacological
treatments using electrophysiological recordings, fluorescent imaging and behavioural tests.
Thomas DeMarse, PhD
Dr. Thomas DeMarse received his MS and PhD in Learning and Memory at Purdue University with
Dr Peter Urcuioli. Dr DeMarse has worked as research fellow in the Biology Department at the
California Institute of Technology and the Biomedical Engineering Department at Georgia Tech
interfacing cortical neurons to computer systems using MEAs in Dr Steve Potter’s Laboratory. He
is currently research scientist in Neural Engineering in the Department of Biomedical Engineering
at the University of Florida. Dr DeMarse’s research interests include basic research into neural
computation including information processing and transmission in combination with MEMs tunnel devices, learning and memory in living neural systems including adapting plasticity for cortical/
hippocampal remodeling (antiepileptogenesis) in epilepsy.
Günther Zeck, PhD
Dr. Zeck completed his PhD in Biophysics at Max Planck Insitute of Biochemsitry in the laboratory
of P. Fromherz, where he stimulated and recorded signals from a synaptically connected neuronal
network on a semiconductor chip.
During his post-doc at the Massachusetts General Hospital and Harvard Medical School in Boston
he started to investigate the coding of vertebrate retinas.
After his return to Germany he now interfaced retina to semiconductor chips (Neurochips) achieving detection and stimulation at high spatial resolution.
Since 2010, Dr. Zeck leads a research group at the NMI at the University Tübingen, where he
extends Neurochip applications to other in vitro preparations, with the long term goal of improving neural implants.
Stanislav Koulchitsky, PhD
Dr. Koulchitsky completed his PhD in biology at the Institute of Physiology (National Academy of
Sciences, Minsk, Belarus), where he studied the role of dorsomedial and ventrolateral medulla in
the regulation of pain sensitivity during endotoxemia. Later, he investigated mechanisms regulating the neuronal excitability in the trigeminal ganglion and the spinal trigeminal nucleus (Institute of Physiology and Experimental Pathophysiology, Erlangen, Germany). In 2008 he joined the
Laboratory of Pharmacology (GIGA Neurosciences, University of Liège). He was involved in the installation of the telemetric system for the registration of neuronal activity from freely moving rats.
His current research fields are the functional wiring of the reward system, the effect of narcotic
drugs on the neuronal populations constituting the reward system, and the basic mechanisms
of addiction.
www.multichannelsystems.com
www.alascience.com