Abstract
This chapter describes the application of electrophysiological techniques in nervous system cultures. The characterization of the membrane properties of cultured neural cells with electrophysiological techniques had already started in 1956 on explant cultures of dorsal root ganglion cells (Crain, 1956) and on glial cells from the cerebellum and midbrain of the kitten (Hild et al., 1958). Subsequently, membrane properties of neurons and glial cells from different areas of the brain were characterized with a combination of electrophyslological, morphological, and immunocytochemical methods.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Baer S. C. and Crain S. M. (1971) Magnetically coupled micromanipulator for use within a sealed chamber. J. Appl. Physiol. 31, 926–929.
Barker J. L. and Mathers D A. (1981) GABA analogues activate channels of different duration on cultured mouse spinal neurons. Science 212, 358–360.
Barker J. L. and McBurney R. N. (1979) GABA and glycine may share the same conductance channel on cultured mammalian neurones. Nature 277, 234–236.
Barker J. L. and Ransom B. R. (1978) Amino acid pharmacology of mammalian central neurones grown in tissue culture. J. Physiol. 280, 331–354.
Bevan S. and Raff M. (1985) Voltage-dependent potassium currents in cultured astrocytes. Nature 315, 229–232.
Bignami A. and Dahl D. (1974) Astrocyte-specific protein and neuroglial differentration An immunofluorescence study with antibodies to the glial fibrillary acidic protein. J. Comp Neurol. 153, 27–37.
Black P. and Kornblith P. L. (1980) Biophysical properties of human astrocytic brain tumor cells in cell culture. J. Cell. Physiol. 105, 565–570.
Bornstein M. B. (1958) Reconstrtuted rat-tail collagen used as a substrate for tissue cultures on covershps in Maximow slides and roller tubes. Lab Invest. 7, 134–137.
Bowman C. L. and Kimelberg H. K. (1984) Excitatory amino acids directly depolarize rat brain astrocytes in primary culture Nature 311, 656–659.
Brown T. H., Perkel D. H., Norris J. C., and Peacock J. H. (1981) Electrotonic structure and specific membrane properties of mouse dorsal root ganglion neurons. J. Neurophysiol 45, 1–15.
Calvet M.-C. (1974) Patterns of spontaneous electrical activity in tissue cultures of mammalian cerebral cortex vs cerebellum, Brain Res 69, 281–295.
Calvet M.-C. and Calvet J. (1979) Horseradish peroxrdase iontophoretic intracellular labelling of cultured Purkinje cells. Brain Res 173, 527–531.
Calvet M.-C., Calvet J., and Camacho-Garcia R (1985) The Purkmje cell dendritic tree: A computer-aided study of its development in the cat and in culture. Brain Res 331, 235–250.
Chiu S Y., Schrager P., and Rltchie J. M. (1984) Neuronal-type Na+ and K+ channels in rabbit cultured Schwann cells. Nature 311, 156–157.
Choi D. W. and Fischbach G. D. (1981) GABA conductance of chick spinal cord and dorsal root ganglion neurons in cell culture. J. Neurophysiol. 45, 605–620.
Corrigall W. A, Crain S. M., and Bornstein M. (1976) Electrophysiological studies of fetal mouse olfactory bulb explants during development of synaptic functions in culture. J. Neurobiol. 7, 521–536
Crain S. M. (1956) Resting and action potentials of cultured embryo chick spinal ganglion cells. J. Neurol. 104, 285–329
Crain S. M. (1973) Microelectrode Recording in Brain Tissue Cultures, in Methods in Physiological Psychology: Bioelectric Recording Techniques: Cellular Processes and Brain Potentials (Thompson R. and Patterson M. M., eds.) Academic, New York.
Crain S and Peterson E. R. (1967) Onset and development of functional interneuronal connections in explants of rat spinal cord-ganglia during maturation in culture. Brain Res. 6, 750–762.
Crain S. M. and Peterson E. R. (1974) Development of specific sensory-evoked synaptic networks in fetal mouse spinal cord-brainstem cultures. Science 188, 275–278.
Dichter M.A (1977) Rat cortical neurons in cell culture: Culture methods, cell morphology, electrophysiology, and synapse formation. Brain Res. 149, 279–293.
Dichter M. A. (1980) Physiological identification of GABA as the inhibitory transmitter for mammalian cortical neurons in cell culture. Brain Res. 190, 111–121.
Dimpfel W., Pierau F.-K., and Haider S. G. (1979) Electrophysiological Studies on the Effects of the Neurotoxin Apamin on Cultured Neurons, in Advances in Cytopharmacology (B. Ceccarelli and F. Clementi, eds.) Raven, New York.
Dreyfus C. F., Gershon M D., and Crain S. M. (1979) Innervation of hippocampal explants by central catecholaminergic neurons in co-cultured fetal mouse brain stem explants. Brain Res. 161, 431–445.
Eng L. F., Vanderhagen J. J., Bignami A., and Gertl B. (1971) An acidic protein isolated from fibrous astrocytes. Brain Res. 28, 351–354.
Fields K. L. (1980) The study of Schwann cells using antigenic markers. Trends Neurosci. 3, 236–238.
Fischbach G. D. and Nelson P. G. (1977) Cell Culture in Neurobiology, in Cellular Biology of Neurons, The Handbook of Physiology (Kandel E. R., ed.) Williams & Wilkins, Baltimore, Maryland.
Fischer G. and Kettenmann H. (1985) Cultured astrocytes form a syncytium after maturation. Exp. Cell Res. 159, 73–279.
Fischer G. and Wieser R. J. (1983) Hormonally Defined Media: A Tool in Cell Biology Springer Verlag, Heidelberg.
Fischer G., Leutz A., and Schachner M. (1982) Cultivation of immature astrocytes in a serum-free, hormonally defined medium. Neurosci. Lett. 29, 97–302.
Fischer G., Kunemund V., and Schachner M. (1986) Neurite outgrowth pattern in cerebellar microexplant cultures are affected by antibodies to the cell surface glycoprotem Ll. J. Neurosci. 6, 605–612
Freschi J.E. (1983) Membrane currents of cultured rat sympathetic neurons under voltage clamp. J. Neurophysiol. 50, 460–478.
Fromm M., Weskamp P. and Hegel U. (1980) Versatile piezoelectric driver for cell puncture. Pflugers Arch. 384, 69–73.
Frosch M. P. and Dichter M. A. (1984) Physiology and pharmacology of olfactory bulb neurons in dissociated cell culture. Brain Res 290, 321–332.
Furshpan E. J., MacLeish P. R., O’Lague P. H., and Potter D. D. (1976) Chemical transmission between rat sympathetic neurons and cardiac myocytes developing in microcultures: Evidence for cholinergic, adrenergic, and dual-function neurones. Proc. Natl. Acad. Sci. USA 73, 4225–4229.
Gahwiler B. H. (1975) The effects of GABA, picrotoxin and bicucullin on the spontaneous bioelectric activity of cultured cerebellar Purkinje cells. Brain Res. 99, 85–95.
Gahwiler B. H. (1976) Spontaneous bioelectric activity of cultured Purkinje cells during exposure to glutamate, glycine, and strychnine. J. Neurobiol. 2, 97–107.
Gahwiler B. H. (1978) Mixed cultures of cerebellum and inferior olive: Generation of complex spikes in Purkinje cells. Brain Res. 145, 168–172.
Gahwiler B. H. (1981) Labeling of neurons within CNS explants by intracellular injection of Lucifer Yellow. J. Neurobiol. 2, 187–191.
Gahwiler B. H. and Bauer W. (1975) Design of a temperature controlled microchamber for electrophysiological experiments in vitro. Separatum Experientia 31, 369.
Gahwiler B. H., Mamoon A. M., Schlapfer W. T., and Tobias C. A. (1972) Effects of temperature on spontaneous bioelectric activity of cultured nerve cells. Brain Res. 40, 527–533.
Gilbert P., Kettenmann H., Orkand R. K., and Schachner M. (1982) Immunocytochemical cell identification in nervous system culture combined with intracellular injection of a blue fluorescing dye (SITS). Neurosci Lett. 334, 123–128.
Gilbert P., Kettenmann H., and Schachner M. (1984) Gamma-aminobutyric acid directly depolarizes cultured oligodendrocytes. J Neurosci. 4, 561–569.
Godfrey E. W., Nelson P. G., Schrier B. K., Breuer A. C., and Ransom B. R. (1975) Neurons from fetal rat brain in a new cell culture system: A multidisciplinary analysis. Brain Res 90, 1–21.
Gross G.W. (1979) Simultaneous single unit recording in vitro with a photoetched laser deinsulated gold multimicroelectrode surface. IEEE Transact. Biomed. Eng. 26, 273–279.
Gross G. W. and Lucas J. H. (1982) Long-term monitoring of spontaneous single unit activity from neuronal monolayer networks cultured on photoetched multielectrode surfaces. J Electrophysiol. Tech. 9, 55–67.
Gross G. W., Rieske E., Kreutzberg G. W., and Meyer A. (1977) A new fixed-array multi-microelectrode system designed for long-term monitoring of extracellular single unit neuronal activity in vitro. Neurosci Lett. 6, 101–105.
Hamill O. P, Marty A., Neher E., Sakmann B., and Sigworth F. J. (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 391, 85–100.
Hicks T P. (1984) The history and development of microiontophoresis in experimental neurobiology. Prog. Neurobiol. 22, 185–240.
Hild W., Chang J. J., and Tasaki I. (1958) Electrical responses of astrocytic gha from the mammalian central nervous system cultivated in vitro. Experientia 14, 220–221.
Hirata H., Slater N. T., and Kimelberg H. K. (1983) Alpha-adrenergic receptor-mediated depolarization of rat neocortical astrocytes in primary culture. Brain Res. 270, 358–362.
Hogue M. J (1947) Human fetal brain cells in tissue culture: Their identification and motility. J Exp. Zool. 106, 85–107.
Hosli L., Andres P. F., and Hosli E (1978) Neuron-glia interactions: Indirect effect of GABA on cultured glial cells. Exp. Brain Res. 33, 425–434.
Hosli L., Hosli E., Landolt H., and Zehntner C. (1981) Efflux of potassium from neurones excited by glutamate and aspartate causes a depolarization of cultured glial cells. Neurosci. Lett. 21, 83–86.
Keilhauer G., Meter D. H., Kuhlmann-Krieg S., Nieke J., and Schachner M. (1985) Astrocyte support incomplete differentiation of an oligodendrocyte precursor cell. EMBO J. 44, 2499–2504.
Kettenmann H. (1985) A reversible decrease in electrical coupling of cultured mouse glial cells induced by superfusion from a micropipette. Neurosci Lett. 59, 85–88.
Kettenmann H. and Orkand R. K. (1983) Intracellular SITS injection dye-uncouples mammalian oligodendrocytes in culture. Neurosci. Lett 39, 21–26.
Kettenmann H. and Schachner M. (1985) Pharmacological properties of GABA, glutamate and aspartate induced depolarizations in cultured astrocytes. J. Neurosci 5, 3295–3301.
Kettenmann H., Orkand R K., Lux H. D., and Schachner M. (1982) Single potassium channel currents in cultured mouse oligodendrocytes. Neurosci. Lett. 32, 41–46
Kettenmann H., Backus H., and Schachner M. (1984a) Aspartate, glutamate and gamma-ammobutyric acid depolarize cultured astrocytes. Neurosci Lett. 52, 25–29.
Kettenmann H, Orkand R K., and Lux H. D (1984b) Some properties of single potassium channels in cultured oligodendrocytes Pflugers Arch. 400, 215–221.
Kettenmann H., Sonnhof U., Camerer H., Kuhlmann S., Orkand R. K, and Schachner M. (1984c) Electrical properties of oligodendrocytes in culture. Pflugers Arch. 401, 324–332
Kettenmann H, Sommer I., and Schachner M (1985) Monoclonal cell surface antibodies do not produce short-term effects on electrical properties of mouse oligodendrocytes in culture Neurosci Lett. 54, 195–199
Kettenmann H., Backus K H., and Schachner M. (1987) GABA opens chloride channels in cultured astrocytes. Brain Res. 404, 1–9.
Kettenmann H., Orkand R. K., and Schachner M. (1983a) Coupling among identified cells in mammalian nervous system cultures. J. Neurosci. 3, 506–516.
Kettenmann H., Sonnhof U., and Schachner M. (1983b) Exclusive potassium dependence of the membrane potential in cultured mouse oligodendrocytes. J. Neurosci. 3, 500–505.
Kettenmann H., Wiennch M., and Schachner M. (1983c) Antibody Ll elected from a micropipette identifies neurons without altering electrical activity. Neurosci Left. 41, 85–90.
Kimelberg H. K., Bowman C., Biddlecome S, and Bourke R. S. (1979) Cation transport and membrane potential properties of primary astroglial cultures from neonatal rat brains. Brain Res. 177, 533–550
Kruse P. F. and Patterson M. K (1973) Tissue Culture: Methods and Application Academic, New York.
Macdonald R. L. and auJ. L. Barker (1981) Neuropharmacology of Spinal Cord Neurons in Primary Dissociated Cell Culture, in Excitable Cells in Tissue Culture (Nelson P. D. and Lieberman M., eds.) Plenum, New York.
MacVicar B. A. (1984) Voltage-dependent calcium channels in glial cells. Science 226, 1345–1347
Mains R. E. and Patterson P H. (1973) Primary cultures of dissociated sympathetic neurons. J. Cell Biol 59, 329–345
Manthorpe M., Engvall E, Ruoslathi E., Longo F, Davis G. E., and Varon S. (1983) Laminin promotes neuritic regeneration from cultured peripheral and central neurons. J, Cell Biol. 97, 1882–1890.
Mathers D. A and Barker J. L (1981) GABA and muscimol open ion channels of different lifetimes on cultured mouse spinal cord cells. Brain Res. 204, 242–247.
McCarthy D. and de Vellis J. (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J, Cell. Biol. 85, 890–902.
Meier D. H., Lagenaur C., and Schachner M. (1982) Immunoselection of oligodendrocytes by magnetic beads. I. Determination of antibody coupling parameters and cell binding conditions. J. Neuroscience Res. 7, 119–134.
Moonen G. and Nelson P G. (1978) Some Physiological Properties of Astrocytes in Primary Cultures, in Dynamic Properties of Glial Cells (Schoffeniels E., Frank G., Hertz L., and Tower D B. L., eds.) Pergamon, London.
Moonen G., Neale E A, Macdonald R. L., Gibbs W, and Nelson P. G. (1982) Cerebellar macroneurons in microexplant cell culture. Methodology, basic electrophysiology, and morphology after horseradish peroxidase injection. Dev Brain Res 5, 59–73.
Neale E. A, Macdonald R. L., and Nelson P. G. (1978) Intracellular horseradish peroxidase injection for correlation of light and electron microscopic anatomy with synaptic physiology of cultured mouse spinal cord neurons. Brain Res. 152, 265–282.
Nelson P. G. (1975) Nerve and muscle cells in culture. Physiol. Rev., 55, 1–61.
Nelson P. G. and Lieberman M. (1981) Excitable Cells in Tissue Culture Plenum, New York
Nelson P G and Peacock J H. (1973) Electrical activity in dissociated cell cultures from fetal mouse cerebellum. Brain Res. 61, 163–174.
Nelson P. G., Ransom B. R., Henkart M., and Bulloch P. N. (1977) Mouse spinal cord in cell culture. IV. Modulation of inhibitory synaptic function. J. Newophysiol. 40, 1178–1186.
O’ Lague P. H., Potter D. D, and Furshpan E. J. (1978a) Studies on rat sympathetic neurons developing in cell culture. I. Growth characteristics and electrophysiological properties. Dev. Biol. 67, 384–403.
O’ Lague P H., Furshpan E. J., and Potter D. D. (1978b) Studies on rat sympathetic neurons developing in cell culture. Dev. Biol. 67, 404–423.
O’ Lague P. H., Potter D. D., and Furshpan E. J (1978c) Studies on rat sympathetic neurons developing in cell culture. III. Cholinergic transmission. Dev Biol. 67, 424–443.
Orkand R. K (1977) Glial Cells, in Cellular Biology of Neurons, The Handbook of Physiology (Kandel E. R., ed.) Williams & Wilkins, Baltimore, Maryland.
Patrick J., Heinemann S., and Schubert D. (1978) Biology of cultured nerve and muscle. Ann. Rev Neurosci. 1, 417–443.
Patterson P H. (1978) Environmental determination of autonomic neurotransmitter functions. Ann Rev. Neurosci 1, 1–17.
Peacock J. H. (1979) Electrophysiology of dissociated hippocampal cultures from fetal mice. Brain Res. 169, 247–260.
Peacock J. H, Nelson P. G., and Goldstone M.W (1973) Electrophysiologic study of cultured neurons dissociated from spinal cords and dorsal root ganglia of fetal mice. Dev Biol. 30, 137–152
Pfeiffer S. E. (1982) Neuroscience Approach to Cell Culture, CRC, Boca Raton, Florida.
Pitman R. M., Tweedle C.D., and Cohen M. J (1972) Branching of central neurons. Intracellular cobalt injection for light and electron microscopy. Science 176, 412–414.
Podulso S. E. and Norton W. T. (1972) Isolation and some chemical properties of oligodendroglia from calf brain. J Newochem. 19, 727–736.
Rall W. (1977) Core Conductor Theory and Cable Properhes of Neurons, in Cellular Biology of Neurons, The Handbook of Physiology (Kandel E. R., ed.) Williams & Wilkins, Baltimore.
Raff M. C., Field K L, Hakomori S-I, Mirsky R., Pruss R. N, and Winter J. (1979) Cell type specific markers for distinguishing and studying neurons and major classes of glial cells in culture. Brain Res. 174, 283–308.
Ransom B. R., and Holz R. W. (1977) Ionic determinants of excitability in cultured mouse dorsal root ganglion and spinal cord cells. Brain Res. 136, 445–453.
Ransom B. R., Neale E., Henkart M., Bullock P. N., and Nelson P. G. (1977a) Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties. J. Neurophysiol 5, 1132–1150.
Ransom B. R., Christian N, Bullock P. N., and Nelson P. G. (1977b) Mouse spinal cord in cell culture. II. Synaptic activity and circuit behaviour. I. Neurophysiol. 5, 1151–1162.
Ransom B. R., Bullock P. N., and Nelson P. G. (1977c) Mouse spinal cord in cell culture. III. Neuronal chemosensitivity and its relationship to synaptic activity. J. Neurophysiology 5, 1163–1177.
Rathjen F. G. and Schachner M (1984) Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion. EMBO J. 3, 1–10.
Sakmann B. and Neher E. (1983) Single Channel Recording Plenum, New York.
Sakman B. and Neher E. (1984) Patch clamp techniques for studying ionx channels in excitable membranes. Ann Rev. Physiol 46, 455–472.
Schachner M (1982a) Cell type-specific surface antigens in the mammalian nervous system J Neurochem 39, 1–8.
Schachner M. (1982b) Glial antigens and the expression of neuroglial phenotypes. Trends Neurosci. 5, 225–229.
Schachner M. (1984) Cell Type-Specific Antigens in the Mammalian Nervous System, in Molecular Biology Approach to the Neurosciences (H. Soreq, ed.) John Wiley, New York.
Schachner M., Kim S. U, and Zehnle R. (1981a) Developmental expression in central and peripheral nervous system of oligodendrocyte cell surface antigens (O antigens) recognized by monoclonal antibodies. Dev Biol 83, 328–338
Schachner M., Sommer I., Lagenaur C and Schmtzer J. (1981b) Monoclonal Antibodies Recognizing Subpopulations of Glial Cells in Mouse Cerebellum, in Monoclonal Antibodies Against Neural Antigens Cold Spring Harbor Reports in the Neurosciences (R. McKay, M. Raff, and L. Reichardt, eds.) Cold Spring Harbor, Long Island.
Schachner M., Sommer I., Lagenaur C and Schnitzer J. (1982) Developmental Expression of Antigenic Markers, in Glial Subclasses, in Neuronal-Glial Cell lnterrelationships (T. Sears, ed.) Life Sciences Research Report, vol. 20, Springer Verlag, Berlin.
Schachner M., Sommer I., Lagenaur C, Schnitzer J., and Berg G. (1983) Antigenic Markers of Glia and Glial Subclasses, in Current Frontiers in Neurobiology Approached Through Cell Culture (S. E. Pfeiffer, ed.) Vol. 2, CRC, Boca Rotan, Florida.
Schlapfer W. T. (1978) Tissue and Organ Culture, in Methods in Neurobiology (R. Lahue, ed) Plenum, New York
Schnitzer J and Schachner M. (1981a) Characterization of isolated mouse cerebellar cell populations in vitro. J. Neuroimmunol. 1, 457–470.
Schnitzer J. and Schachner M. (1981b) Developmental expression of cell type-specific markers in mouse cerebellar cells in vitro. J Neuroimmunol. 1, 471–487.
Shrager P., Chiu S. Y., and Ritchie J. M. (1985) Voltage-dependent sodium and potassium channels in mammalian cultured Schwann cells. Proc. Natl. Acad. Sci. USA 82, 948–952.
Sommer I. and Schachner M. (1981) Monoclonal antibodies (01 to 04) to oligodendrocyte cell surfaces: An immunocytological study in the central nervous system. Dev Biol. 83, 311–327.
Sommer I. and Schachner M. (1982) Cells that are 04 antigen-positive and 01 antigen-negative differentiate into 01 antigen-positive oligodendrocytes. Neurosci Lett 29, 183–188.
Sonnhof U. and Schachner M (1986) Single voltage dependent K+-channels in cultured astrocytes. Neurosci Lett. 64, 241–246
Sonnhof U., Forderer R., Schneider W., and Kettenmann H. (1982) Cell puncturing with a step motor driven manipulator with simultaneous measurement of displacement. Pflugers Arch 392, 295–300.
Stewart W. W. (1981) Lucifer dyes-highly fluorescent dyes for biological tracing Nature 292, 17–24.
Walz W., Wuttke W., and Hertz L. (1984) Astrocytes in primary cultures. Membrane potential characteristics reveal exclusive potassium conductance and potassium accumulator properties. Brain Res. 292, 367–374.
Wardell W. M. (1966) Electrical and pharmacological properties of mammalian neuroglial cells in tissue-culture. Proc. Roy Soc. Lond. (Bio1.) 165, 326–361
Weber A. and Schachner M. (1984) Maintenance of immunocytologically identified Purkinje cells from mouse cerebellum in monolayer culture. Brain Res. 311, 119–130.
Weir D. M. (1978) Handbook of Experiemental immunology vol. I, Immunochemistry Blackwell Scientific, Oxford.
Werz W. and Schachner M. (1987) Extracellular matrix molecules promote neuronal outgrowth of cerebelar cells (in preparation).
Wienrich M. and Kettenmann H. (1984) Intrazellulare Farbstoffe zur Markierung einzelner neuraler Zellen in Kultur. Kontukte (Darmstadt) 3, 30–44.
Yavin E. and Yavin Z. (1974) Attachment and culture of dissociated cells from rat embryo cerebral hemispheres on poly-L-lysine coated surface. J. Cell Biol. 62, 540–546
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1988 The Humana Press Inc.
About this protocol
Cite this protocol
Kettenmann, H. (1988). Electrophysiological Methods Applied in Nervous System Cultures. In: Boulton, A.A., Baker, G.B., Walz, W. (eds) The Neuronal Microenvironment. Neuromethods, vol 9. Humana Press. https://doi.org/10.1385/0-89603-115-2:493
Download citation
DOI: https://doi.org/10.1385/0-89603-115-2:493
Publisher Name: Humana Press
Print ISBN: 978-0-89603-115-9
Online ISBN: 978-1-59259-614-0
eBook Packages: Springer Protocols