Summary
Quantitative analysis of neuronal firing patterns was used to study the effects of chronic suppression of bioelectric activity (BEA) on functional development in primary cultures of fetal rat cerebral cortex. BEA was monitored with extracellular electrodes in active control cultures or, after return to control medium, in cultures chronically silenced with tetrodotoxin (TTX) at around 7, 14, 21 and 42 days in vitro. Spike trains of single neurons lasting up to 25 min duration were analyzed using a previously published set of computer programs. In control cultures, the main developmental trends seen in a previous study could be replicated. After development in the presence of TTX, activity levels were increased at all ages, and a high incidence was found of a single firing pattern characterized by stereotyped burst firing, while showing a low minute order variability in firing rate and low dependencies between successive intervals; conversely, the incidence of variable/non-burst firing was decreased relative to untreated cultures. The former firing pattern (i.e. non-variable bursting) could also be produced through acute addition of the GABA (A)-antagonist picrotoxin to control cultures, and resembled interictal burst firing observed in models of chronic epilepsy in vivo. These similarities suggest that chronic silencing of the cultures may have resulted in a functional disinhibition of the neuronal network; such disinhibition might be related to the increased cell death which we observed with chronic TTX-treatment in the same cultures.
Similar content being viewed by others
References
Archer SM, Dubin MW, Stark LA (1982) Abnormal development of kitten retino-geniculate connectivity in the absence of action potentials. Science 217:743–745
Baker RE, Corner MA, Habets AMMC (1984) Effects of chronic suppression of bioelectric activity on the development of sensory ganglion evoked responses in spinal cord explants. J Neurosci 4:1187–1192
Baker RE, Corner MA, Lammertse T, Fürth E (1986) Some functional effects of suppressing bioelectric activity in mouse spinal cord/dorsal root ganglion explants. Exp Neurol 94:426–430
Black IB, Adler JE, Dreyfus CF, Jonakait GM, Katz DM, LaGamma EF, Markey KM (1984) Neurotransmitter plasticity at the molecular level. Science 225:1266–1270
Bornstein M (1958) Rat tail collagen as substrate for tissue cultures on coverslips in Maximow slides and in roller tubes. Lab Invest 7:134–137
Brenneman DE, Nelson PG (1985) Neuronal development in culture: role of electrical activity. In: Bottenstein JE, Sato G (eds) Cell culture in the neurosciences. Plenum Press, New York, pp 289–316
Brodie C, Brody M, Sampson SR (1989) Characterization of the relation between sodium channels and electrical activity in cultured rat skeletal myotubes: regulatory aspects. Brain Res (in press)
Brunk CF, Jones KC, James TW (1979) Assay for nanogram quantitative of DNA in cellular homogenates. Anal Biochem 92:497–500
Calvet M-C (1974) Patterns of spontaneous electrical activity in tissue cultures of mammalian cerebral cortex vs. cerebellum. Brain Res 69:281–295
Cohan CS, Kater SB (1986) Suppression of neurite elongation and growth cone dynamics by electrical activity. Science 232:1638–1640
Corner MA (1985) Ontogeny of brain sleep mechanisms. In: McGinty DJ, Drucker-Colin RR, Morison AR, Parmeggiani PL (eds) Brain mechanisms of sleep. Raven Press, New York, pp 175–198
Crain SM (1976) Neurophysiologic studies in tissue culture. Raven Press, New York
Davenne D, Frégnac Y, Imbert M, Adrien J (1989) Lesion of the PGO pathways in the kitten. II. Impairment of physiological and morphological maturation of the lateral geniculate nucleus. Dev Brain Res 485:267–277
Dichter MA (1980) Physiological identification of GABA as the inhibitory transmitter for mammalian cortical neurons in cell culture. Brain Res 190:111–121
Droge MH, Gross GW, Hightower MH, Czisny LE (1986) Multi-electrode analysis of coordinated, multisite, rhythmic bursting in cultured CNS monolayer networks. J Neurosci 6:1583–1592
Habets AMMC, Van Dongen AMJ, Van Huizen F, Corner MA (1987) Spontaneous neuronal firing patterns in fetal rat cortical networks during development in vitro: a quantitative analysis. Exp Brain Res 69:43–52
Laing NG (1982) Motor projection patterns to the hind limb of normal and paralysed chick embryos. J Embryol Exp Morphol 72:269–286
Lipton SA (1986) Blockade of electrical activity promotes the death of mammalian retinal ganglion cells in culture. Proc Natl Acad Sci USA 83:9774–9778
Löscher W, Schwark WS (1987) Further evidence for abnormal GABAergic circuits in amygdala-kindled rats. Brain Res 420:385–390
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Maderdrut JL, Oppenheim RW, Prevette D (1988) Enhancement of naturally occurring cell death in the sympathetic and parasympathetic ganglia of the chicken embryo following blockade of ganglionic transmission. Brain Res 444:189–194
Ramakers GJA (1987) Changes in the firing patterns of spontaneously active neurons in primary cultures of dissociated rat cerebral cortex after chronic blockade of bioelectric activity. Neuroscience 22 Suppl:401
Ramakers GJA, Corner MA (1987) Effects of alterations in the level of spontaneous bioelectric activity on the development of firing patterns in cultures of dissociated cerebral cortex. Soc Neurosci Abstr 17:1426
Ribak CE (1985) Axon terminals of GABAergic chandelier cells are lost at epileptic foci. Brain Res 326:251–260
Ribak CE, Hunt CA, Bakay RAE, Oertel WH (1986) A decrease in the number of GABAergic somata is associated with the preferential loss of GABAergic terminals at epileptic foci. Brain Res 363:78–90
Romijn HJ, Van Huizen F, Wolters PS (1984) Towards an improved serum-free, chemically defined medium for longterm culturing of cerebral cortex tissue. Neurosci Biobehav Rev 8:301–334
Ruijter JM, Baker RE, De Jong BM, Romijn HJ (1989) Chronic blockade of bioelectric activity in neocortical rat cortex grown in vitro: morphological effects. Int J Dev Neurosci (in press)
Shatz CJ, Stryker MP (1988) Prenatal tetrodotoxin infusion blocks segregation of retinogeniculate afferents. Science 242:87–89
Sherman SJ, Catterall WA (1984) Electrical activity and cytosolic calcium regulate levels of tetrodotoxin-sensitive sodium channels in cultured rat muscle cells. Proc Natl Acad Sci USA 81:262–266
Siegel S, Castellan NJ (1988) Nonparametric statistics for the behavioural sciences. McGraw-Hill, New York
SPSSX Inc. (1983) SPSSX user's guide: a complete guide to SPSSX language and operations. McGraw-Hill, New York
Tsuru N (1985) Neuronal firing pattern following amygdaloid kindling in unrestrained rats. Epilepsia 26:488–492
Van Huizen F, Romijn HJ, Habets AMMC (1985) Synaptogenesis in rat cerebral cortex cultures is affected during chronic blockade of spontaneous bioelectric activity by tetrodotoxin. Dev Brain Res 19:67–80
Van Huizen F, Romijn HJ, Habets AMMC (1987) Tetrodotoxin enhances initial neurite outgrowth from fetal rat cerebral cortex cells in vitro. Brain Res 408:271–274
Wyler AR, Fetz EE, Ward Jr AA (1975) Firing patterns of epileptic and normal neurons in the chronic alumina focus in undrugged monkeys during different behavioural states. Brain Res 98:1–20
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ramakers, G.J.A., Corner, M.A. & Habets, A.M.M.C. Development in the absence of spontaneous bioelectric activity results in increased stereotyped burst firing in cultures of dissociated cerebral cortex. Exp Brain Res 79, 157–166 (1990). https://doi.org/10.1007/BF00228885
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00228885