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Chapter 6: A Tour of the Cell • All organisms are made of cells • The cell is the simplest collection of matter that can live • Cell structure is correlated to cellular function 10 µm Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Microscopy • Light microscopes (LMs) – Pass visible light through a specimen and magnify cellular structures with lenses • Cells were discovered by ________in 1665 but their ultrastructure was largely unknown until the development of the ____________ in the 1950s. • Electron microscopes (EMs) – Focus a beam of electrons through a specimen (TEM) or onto its surface (SEM) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Three scanning electron microscopes (SEM) and one transmission electron microscope (TEM) provide ultra high-resolution microscopy (magnification to 1,000,000x) to identify extremely small particles. All are equipped with energy dispersive x-ray spectroscopy (EDS) to provide elemental analysis. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 10 m Human height Length of some nerve and muscle cells 0.1 m Chicken egg 1 cm Light microscope Frog egg 100 µm 10 µ m 1µm 100 nm Most plant and Animal cells Nucleus Most bacteria Mitochondrion Smallest bacteria Viruses 10 nm Ribosomes Electron microscope 1 mm Electron microscope Unaided eye • Different types of microscopes can be used to visualize different sized cellular structures 1m Proteins 1 nm Lipids Small molecules Figure 6.2 0.1 nm Atoms Measurements 1 centimeter (cm) = 102 meter (m) = 0.4 inch 1 millimeter (mm) = 10–3 m 1 micrometer (µm) = 10–3 mm = 10–6 m 1 nanometer (nm) = 10–3 mm = 10–9 m Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings – Use different methods for enhancing visualization of cellular structures TECHNIQUE RESULT (a) Brightfield (unstained specimen). Passes light directly through specimen. Unless cell is naturally pigmented or artificially stained, image has little contrast. [Parts (a)–(d) show a human cheek epithelial cell.] 50 µm (b) Brightfield (stained specimen). Staining with various dyes enhances contrast, but most staining procedures require that cells be fixed (preserved). (c) Phase-contrast. Enhances contrast in unstained cells by amplifying variations in density within specimen; especially useful for examining living, unpigmented cells. Figure 6.3 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings (d) Differential-interference-contrast (Nomarski). Like phase-contrast microscopy, it uses optical modifications to exaggerate differences in density, making the image appear almost 3D. (e) Fluorescence. Shows the locations of specific molecules in the cell by tagging the molecules with fluorescent dyes or antibodies. These fluorescent substances absorb ultraviolet radiation and emit visible light, as shown here in a cell from an artery. 50 µm (f) Confocal. Uses lasers and special optics for “optical sectioning” of fluorescently-stained specimens. Only a single plane of focus is illuminated; out-of-focus fluorescence above and below the plane is subtracted by a computer. A sharp image results, as seen in stained nervous tissue (top), where nerve cells are green, support cells are red, and regions of overlap are yellow. A standard fluorescence micrograph (bottom) of this relatively thick tissue is blurry. 50 µm Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The scanning electron microscope (SEM) – Provides for detailed study of the surface of a specimen TECHNIQUE RESULTS 1 µm Cilia (a) Scanning electron microscopy (SEM). Micrographs taken with a scanning electron microscope show a 3D image of the surface of a specimen. This SEM shows the surface of a cell from a rabbit trachea (windpipe) covered with motile organelles called cilia. Beating of the cilia helps move inhaled debris upward toward the throat. Figure 6.4 (a) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The transmission electron microscope (TEM) – Provides for detailed study of the internal ultrastructure of cells Longitudinal section of cilium (b) Transmission electron microscopy (TEM). A transmission electron microscope profiles a thin section of a specimen. Here we see a section through a tracheal cell, revealing its ultrastructure. In preparing the TEM, some cilia were cut along their lengths, creating longitudinal sections, while other cilia were cut straight across, creating cross sections. Figure 6.4 (b) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cross section of cilium 1 µm Interactive Question a. Define cytology b. What do cell biologists use a TEM to study? c. What does and SEM show best? d. What advantages does light microscopy have over TEM and SEM? Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Isolating Organelles by Cell Fractionation • Cell fractionation – Takes cells apart and separates the major organelles from one another • The centrifuge – Is used to fractionate cells into their component parts Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The process of cell fractionation APPLICATION Cell fractionation is used to isolate (fractionate) cell components, based on size and density. TECHNIQUE First, cells are homogenized in a blender to break them up. The resulting mixture (cell homogenate) is then centrifuged at various speeds and durations to fractionate the cell components, forming a series of pellets. RESULTS In the original experiments, the Homogenization Tissue cells 1000 g Homogenate (1000 times the force of gravity) Differential centrifugation 10 min Supernatant poured into next tube 20,000 g 20 min Pellet rich in nuclei and cellular debris 80,000 g 60 min Pellet rich in researchers used microscopy to identify the organelles mitochondria in each pellet, establishing a baseline for further experiments. (and chloroplasts if cells In the next series of experiments, researchers used are from a Pellet rich in biochemical methods to determine the metabolic functions plant) “microsomes” (pieces of associated with each type of organelle. Researchers currently plasma membranes and use cell fractionation to isolate particular organelles in order to cells’ internal study further details of their function. Figure 6.5 membranes) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 150,000 g 3 hr Pellet rich in ribosomes What do all cells have in common? 1. _____, 2. _____, 3. _____, 4. _____ • Prokaryotic cells do not contain a _______ and have their DNA located in a region called the _________ • Eukaryotic cells – Contain a true nucleus, bounded by a membranous nuclear envelope – Are generally quite a bit ______(in size) when compared to prokaryotic cells Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Pili: attachment structures on the surface of some prokaryotes Nucleoid: region where the cell’s DNA is located (not enclosed by a membrane) Ribosomes: organelles that synthesize proteins Bacterial chromosome (a) A typical rod-shaped bacterium Plasma membrane: membrane enclosing the cytoplasm Cell wall: rigid structure outside the plasma membrane Capsule: jelly-like outer coating of many prokaryotes 0.5 µm Flagella: locomotion organelles of some bacteria Figure 6.6 A, B Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings (b) A thin section through the bacterium Bacillus coagulans (TEM) The logistics of carrying out cellular metabolism sets limits on the size of cells • A smaller cell – Has a higher surface to volume ratio, which facilitates the exchange of materials into and out of the cell Surface area increases while total volume remains constant 5 1 1 Total surface area (height width number of sides number of boxes) 6 150 750 Total volume (height width length number of boxes) 1 125 125 Surface-to-volume ratio (surface area volume) 6 12 6 Figure 6.7 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The plasma membrane – Functions as a selective barrier – Allows sufficient passage of nutrients and waste Outside of cell Carbohydrate side chain Hydrophilic region Inside of cell 0.1 µm Hydrophobic region Figure 6.8 A, B (a) TEM of a plasma membrane. The plasma membrane, here in a red blood cell, appears as a pair of dark bands separated by a light band. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Hydrophilic region Phospholipid Proteins (b) Structure of the plasma membrane A panoramic view: a eukaryotic cell • A animal cell ENDOPLASMIC RETICULUM (ER) Rough ER Smooth ER Nuclear envelope Nucleolus NUCLEUS Chromatin Flagelium Plasma membrane Centrosome CYTOSKELETON Microfilaments Intermediate filaments Ribosomes Microtubules Microvilli Golgi apparatus Peroxisome Figure 6.9 Mitochondrion Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Lysosome In animal cells but not plant cells: Lysosomes Centrioles Flagella (in some plant sperm) A panoramic view: a eukaryotic cell II • A plant cell Nuclear envelope Nucleolus Chromatin NUCLEUS Centrosome Rough endoplasmic reticulum Smooth endoplasmic reticulum Ribosomes (small brwon dots) Central vacuole Tonoplast Golgi apparatus Microfilaments Intermediate filaments CYTOSKELETON Microtubules Mitochondrion Peroxisome Plasma membrane Chloroplast Cell wall Plasmodesmata Wall of adjacent cell Figure 6.9 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings In plant cells but not animal cells: Chloroplasts Central vacuole and tonoplast Cell wall Plasmodesmata The Nucleus: Genetic Library of the Cell • The nucleus contains most of the genes in the eukaryotic cell Nucleus 1 µm Nucleolus Chromatin Nucleus Nuclear envelope: Inner membrane Outer membrane Nuclear pore • The nuclear envelope encloses the nucleus, separating its contents from the cytoplasm Pore complex Rough ER Surface of nuclear envelope. 1 µm Ribosome 0.25 µm Close-up of nuclear envelope Nuclear lamina (TEM). Pore complexes (TEM). Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Figure 6.10 Ribosomes: Protein Factories in the Cell • Ribosomes are particles made of ribosomal RNA and protein, and they carry out protein synthesis Ribosomes Cytosol Free ribosomes Bound ribosomes Large subunit 0.5 µm TEM showing ER and ribosomes Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Small subunit Diagram of a ribosome Interactive Question • How does the nucleus control protein synthesis in cytoplasm? Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The ER membrane • Accounts for more than half the total membrane in many eukaryotic cells • Is continuous with the nuclear envelope • 2 kinds of ER • Smooth: lacks ribosomes • Rough: contains ribosomes Figure 6.12 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Smooth ER Rough ER Nuclear envelope ER lumen Cisternae Ribosomes Transitional ER Transport vesicle Smooth ER Rough ER 200 µm Functions of Smooth ER • The smooth ER – Synthesizes lipids – Metabolizes carbohydrates – Stores calcium – Detoxifies poison Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Functions of Rough ER • The rough ER – Has bound ribosomes – Produces proteins and membranes, which are distributed by transport vesicles Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Golgi Apparatus: Shipping and Receiving Center • The Golgi apparatus – Receives many of the transport vesicles produced in the rough ER – Consists of flattened membranous sacs called cisternae • Functions of the Golgi apparatus include – Modification of the products of the rough ER – Manufacture of certain macromolecules Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Functions of the Golgi apparatus Golgi apparatus cis face (“receiving” side of Golgi apparatus) 1 Vesicles move 2 Vesicles coalesce to 6 Vesicles also form new cis Golgi cisternae from ER to Golgi transport certain Cisternae proteins back to ER 3 Cisternal maturation: Golgi cisternae move in a cisto-trans direction Figure 6.13 5 Vesicles transport specific proteins backward to newer Golgi cisternae 4 Vesicles form and leave Golgi, carrying specific proteins to other locations or to the plasma membrane for secretion trans face (“shipping” side of Golgi apparatus) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 0.1 0 µm TEM of Golgi apparatus Lysosomes: Digestive Compartments • A lysosome – Is a membranous sac of hydrolytic enzymes – Can digest all kinds of macromolecules Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Lysosomes carry out intracellular digestion by – Phagocytosis Nucleus 1 µm Lysosome Lysosome contains active hydrolytic enzymes Food vacuole fuses with lysosome Hydrolytic enzymes digest food particles Digestive enzymes Lysosome Plasma membrane Digestion Food vacuole Figure 6.14 A Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings (a) Phagocytosis: lysosome digesting food • Autophagy Lysosome containing two damaged organelles 1µm Mitochondrion fragment Peroxisome fragment Lysosome fuses with vesicle containing damaged organelle Hydrolytic enzymes digest organelle components Lysosome Vesicle containing damaged mitochondrion Digestion (b) Autophagy: lysosome breaking down damaged organelle Figure 6.14 B Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Vacuoles: Diverse Maintenance Compartments • A plant or fungal cell – May have one or several vacuoles • Food vacuoles – Are formed by phagocytosis • Contractile vacuoles – Pump excess water out of protist cells Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Central vacuoles Central vacuole – Are found in plant cells Cytosol Tonoplast – Hold reserves of important organic compounds and water Figure 6.15 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Nucleus Central vacuole Cell wall Chloroplast 5 µm • Relationships among organelles of the endomembrane system 1 Nuclear envelope is connected to rough ER, which is also continuous with smooth ER Nucleus Rough ER 2 Membranes and proteins produced by the ER flow in the form of transport vesicles to the Golgi Smooth ER cis Golgi Nuclear envelop 3 Golgi pinches off transport Vesicles and other vesicles that give rise to lysosomes and Vacuoles Plasma membrane trans Golgi 4 Lysosome available for fusion with another vesicle for digestion Figure 6.16 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 5 Transport vesicle carries 6 proteins to plasma membrane for secretion Plasma membrane expands by fusion of vesicles; proteins are secreted from cell • Concept 6.5: Mitochondria and chloroplasts change energy from one form to another • Mitochondria – Are the sites of cellular respiration • Chloroplasts – Found only in plants, are the sites of photosynthesis Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Mitochondria are enclosed by two membranes – Are found in nearly all eukaryotic cells and have smooth outer membrane – An inner membrane folded into cristae Mitochondrion Intermembrane space Outer membrane Free ribosomes in the mitochondrial matrix Inner membrane Cristae Matrix Figure 6.17 Mitochondrial DNA Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 100 µm Chloroplasts: Capture of Light Energy • The chloroplast contains chlorophyll • Is a specialized member of a family of closely related plant organelles called plastids; this is where photosythesis takes place • Are found in leaves and other green organs of plants and in algae Chloroplast Ribosomes Stroma Inner and outer membranes Granum Chloroplast DNA 1 µm Thylakoid Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Chloroplast structure includes – Thylakoids, membranous sacs – Stroma, the internal fluid Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Peroxisomes: Oxidation • Peroxisomes – Convert hydrogen peroxide to water (because H2O2 is a toxic by-product Chloroplast Peroxisome Mitochondrion Figure 6.19 1 µm Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The cytoskeleton – Is a network of fibers extending throughout the cytoplasm and gives mechanical support to the cell Microtubule Figure 6.20 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 0.25 µm Microfilaments – Is involved in cell motility, which utilizes motor proteins ATP Vesicle Receptor for motor protein Motor protein Microtubule (ATP powered) of cytoskeleton (a) Motor proteins that attach to receptors on organelles can “walk” the organelles along microtubules or, in some cases, microfilaments. Vesicles Microtubule 0.25 µm Figure 6.21 A, B (b) Vesicles containing neurotransmitters migrate to the tips of nerve cell axons via the mechanism in (a). In this SEM of a squid giant axon, two vesicles can be seen moving along a microtubule. (A separate part of the experiment provided the evidence that they were in fact moving.) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • There are three main types of fibers that make up the cytoskeleton Table 6.1 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Microtubules • Microtubules – Shape the cell – Guide movement of organelles – Help separate the chromosome copies in dividing cells Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The centrosome – Is considered to be a “microtubule-organizing center” and contains a pair of centrioles Centrosome Microtubule Centrioles 0.25 µm Figure 6.22 Longitudinal section of one centriole Microtubules Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cross section of the other centriole Cilia and Flagella • Cilia and flagella – Contain specialized arrangements of microtubules – Are locomotor appendages of some cells Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Flagella beating pattern (a) Motion of flagella. A flagellum usually undulates, its snakelike motion driving a cell in the same direction as the axis of the flagellum. Propulsion of a human sperm cell is an example of flagellatelocomotion (LM). Direction of swimming Figure 6.23 A 1 µm Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Ciliary motion (b) Motion of cilia. Cilia have a backand-forth motion that moves the cell in a direction perpendicular to the axis of the cilium. A dense nap of cilia, beating at a rate of about 40 to 60 strokes a second, covers this Colpidium, a freshwater protozoan (SEM). Figure 6.23 B 15 µm Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Cilia and flagella share a common ultrastructure Outer microtubule doublet Dynein arms 0.1 µm Central microtubule Outer doublets cross-linking proteins inside Microtubules Radial spoke Plasma membrane Basal body (b) 0.5 µm (a) 0.1 µm Triplet (c) Figure 6.24 A-C Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cross section of basal body Plasma membrane • The protein dynein – Is responsible for the bending movement of cilia and flagella Microtubule doublets ATP Dynein arm (a) Powered by ATP, the dynein arms of one microtubule doublet grip the adjacent doublet, push it up, release, and then grip again. If the two microtubule doublets were not attached, they would slide relative to each other. Figure 6.25 A Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings ATP Outer doublets cross-linking proteins Anchorage in cell (b) In a cilium or flagellum, two adjacent doublets cannot slide far because they are physically restrained by proteins, so they bend. (Only two of the nine outer doublets in Figure 6.24b are shown here.) Figure 6.25 B Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 1 3 2 (c) Localized, synchronized activation of many dynein arms probably causes a bend to begin at the base of the Cilium or flagellum and move outward toward the tip. Many successive bends, such as the ones shown here to the left and right, result in a wavelike motion. In this diagram, the two central microtubules and the cross-linking proteins are not shown. Figure 6.25 C Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Microfilaments (Actin Filaments) • Microfilaments – Are built from Microvillus molecules of the protein actin and Plasma membrane are found in microvilli Microfilaments (actin filaments) Intermediate filaments Figure 6.26 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 0.25 µm • Microfilaments that function in cellular motility – Contain the protein myosin in addition to actin Muscle cell Actin filament Myosin filament Myosin arm Figure 6.27 A (a) Myosin motors in muscle cell contraction. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Amoeboid movement – Involves the contraction of actin and myosin filaments Cortex (outer cytoplasm): gel with actin network Inner cytoplasm: sol with actin subunits Extending pseudopodium Figure 6.27 B (b) Amoeboid movement Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Cytoplasmic streaming – Is another form of locomotion created by microfilaments Nonmoving cytoplasm (gel) Chloroplast Streaming cytoplasm (sol) Parallel actin filaments Figure 6.27 C (b) Cytoplasmic streaming in plant cells Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cell wall Intermediate Filaments • Intermediate filaments – Support cell shape – Fix organelles in place Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cell Walls of Plants – The cell wall is an extracellular structure of plant cells that distinguishes them from animal cells – Are made of cellulose fibers embedded in other polysaccharides and protein – May have multiple layers Central vacuole of cell Plasma membrane Secondary cell wall Primary cell wall Central vacuole of cell Middle lamella 1 µm Central vacuole Cytosol Plasma membrane Plant cell walls Figure 6.28 Plasmodesmata Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Interactive Question • If you sketched two adjacent plant cells, could you show the location of the primary and secondary cell walls and the middle lamella? Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Extracellular Matrix (ECM) of Animal Cells • Animal cells lack cell walls and are covered by an elaborate matrix, the ECM, which is made up of glycoproteins and other macromolecules EXTRACELLULAR FLUID Collagen A proteoglycan complex Polysaccharide molecule Carbohydrates Core protein Fibronectin Plasma membrane Integrin Integrins Microfilaments Figure 6.29 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings CYTOPLASM Proteoglycan molecule • Functions of the ECM include – Support – Adhesion – Movement – Regulation Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Plants: Plasmodesmata • Plasmodesmata – Are channels that perforate plant cell walls Cell walls Interior of cell Interior of cell Figure 6.30 0.5 µm Plasmodesmata Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Plasma membranes Animals: Tight Junctions, Desmosomes, and Gap Junctions • In animals, there are three types of intercellular junctions – Tight junctions – Desmosomes – Gap junctions Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Types of intercellular junctions in animals TIGHT JUNCTIONS Tight junction Tight junctions prevent fluid from moving across a layer of cells 0.5 µm At tight junctions, the membranes of neighboring cells are very tightly pressed against each other, bound together by specific proteins (purple). Forming continuous seals around the cells, tight junctions prevent leakage of extracellular fluid across A layer of epithelial cells. DESMOSOMES Desmosomes (also called anchoring junctions) function like rivets, fastening cells Together into strong sheets. Intermediate Filaments made of sturdy keratin proteins Anchor desmosomes in the cytoplasm. Tight junctions Intermediate filaments Desmosome Gap junctions Space between Plasma membranes cells of adjacent cells 1 µm Extracellular matrix Gap junction Figure 6.31 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 0.1 µm GAP JUNCTIONS Gap junctions (also called communicating junctions) provide cytoplasmic channels from one cell to an adjacent cell. Gap junctions consist of special membrane proteins that surround a pore through which ions, sugars, amino acids, and other small molecules may pass. Gap junctions are necessary for communication between cells in many types of tissues, including heart muscle and animal embryos. The Cell: A Living Unit Greater Than the Sum of Its Parts 5 µm • Cells rely on the integration of structures and organelles in order to function Figure 6.32 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings