Experimental Brain Research

, Volume 69, Issue 1, pp 43–52 | Cite as

Spontaneous neuronal firing patterns in fetal rat cortical networks during development in vitro: a quantitative analysis

  • A. M. M. C. Habets
  • A. M. J. Van Dongen
  • F. Van Huizen
  • M. A. Corner
Article

Summary

The development of spontaneous bioelectric activity (SBA) was studied in dissociated occipital cortex cultures prepared from 19 day old rat fetuses. All cultures, recorded one per diem from 5 to 33 days in vitro (div), showed SBA. Computer analysis of 76 extracellularly recorded single unit spike trains was carried out after selection on the basis of stationarity criteria. Statistically significant developmental trends were found in (i) interspike interval dependencies and (ii) fluctuations in mean firing rate, on the order of a minute or longer. The highly dependent firing patterns, including stereotyped bursting, were present mostly in the 9–12 div group, whereas minute-to-minute fluctuations in the intensity of firing were considerably more pronounced in the oldest group (22–33 div) than in the younger cultures. In addition, firing categories defined on the basis of factor-analysis revealed that such fluctuations were almost exclusively to be found in neurons which fired in a pronounced ‘burst’, rather than a relatively continuous fashion. Only a few mature appearing synaptic structures were observed electron microscopically prior to 12 div, but increased steadily in number thereafter. No cultures prior to 14 div, but all cultures older than this, stained positively for the presence of glutamic acid decarboxylase. An extensive immunoreactive, putative GABAergic, network was present by three weeks in vitro.

Key words

Spike train analysis Spontaneous activity Primary culture Neuronal development Occipital cortex Rat 

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References

  1. Calvet M-C (1974) Patterns of spontaneous electrical activity in tissue cultures of mammalian cerebral cortex vs. cerebellum. Brain Res 69: 281–295Google Scholar
  2. Connors BW, Benardo LS, Prince DA (1983) Coupling between neurons of the developing rat neocortex. J Neurosci 3: 773–782Google Scholar
  3. Connors BW, Gutnick MJ, Prince DA (1982) Electrophysiological properties of neocortical neurons in vitro. J Neurophysiol 48: 1302–1320Google Scholar
  4. Corner MA (1985) Ontogeny of brain sleep mechanisms. In: McGinty DJ et al. (eds) Brain mechanisms of sleep. Raven Press, New York, pp 175–197Google Scholar
  5. Corner MA, Crain SM (1972) Patterns of spontaneous bioelectric activity during maturation in culture of fetal rodent medulla and spinal cord tissues. J Neurobiol 3: 25–45Google Scholar
  6. Coyle JT, Enna SJ (1976) Neurochemical aspects of the ontogenesis of GABAergic neurons in the rat brain. Brain Res 111: 119–133Google Scholar
  7. Cox DR, Lewis PAW (1966) The statistical analysis of series of events. Methuen, LondonGoogle Scholar
  8. De Kwaadsteniet JW (1982) Statistical analysis and stochastic modeling of neuronal spike-train activity. Math Biosci 60: 17–71Google Scholar
  9. Dichter MA (1978) Rat cortical neurons in cell culture: culture methods, cell morphology, electrophysiology and synapse formation. Brain Res 149: 279–293Google Scholar
  10. Dichter MA (1980) Physiological identification of GABA as the inhibitory transmitter for mammalian cortical neurons in cell culture. Brain Res 190: 111–121Google Scholar
  11. Droge MH, Gross GW, Hightower MH, Czisny LE (1986) Multielectrode analysis of coordinated, multisite, rhythmic bursting in cultured CNS monolayer networks. J Neurosci 6: 1583–1592Google Scholar
  12. Dyson SE, Jones DG (1976) The morphological categorisation of developing synaptic junctions. Cell Tiss Res 167: 363–371Google Scholar
  13. Harter HL (1980) Modified asymptotic formulas for critical values of the Kolmogorov test statistic. Am Statist 34: 110–111Google Scholar
  14. Jackson MB, Lecar H, Brenneman DE, Fitzgerald S, Nelson P (1982) Electrical development in spinal cord cell culture. J Neurosci 2: 1052–1061Google Scholar
  15. O'Brien RJ, Fischbach GD (1986) Excitatory synaptic transmission between interneurons and motoneurons in chick spinal cord cell cultures. J Neurosci 6: 3284–3289Google Scholar
  16. Nakahama H, Yamamoto M, Ishhi N, Fujii H, Aya K (1977) Dependency as a measure to estimate the order and the values of Markov processes. Biol Cybern 25: 209–226Google Scholar
  17. Neale EA, Oertel WH, Bowers LM, Weise VK (1983) Glutamate decarboxylase immunoreactivity and gamma-[3H]aminobutyric acid accumulation within the same neurons in dissociated cell cultures of cerebral cortex. J Neurosci 3: 376–382Google Scholar
  18. Perkel OH, Gerstein GL, Moore GP (1967) Neuronal spike trains. I. The single spike train. Bioph J 7: 391–418Google Scholar
  19. Provine RP (1976) Development of function in nerve nets. In: Fentress JC (ed) Simpler networks and behavior. Sinauer, Sunderland Mass, pp 203–220Google Scholar
  20. Purpura DP, Housepian EM (1961) Morphological and physiological properties of chronically isolated immature neocortex. Exp Neurol 4: 377–401Google Scholar
  21. Romijn HJ, Mud M, Habets AMMC, Wolters PS (1981) A quantitative electron microscopic study on synapse formation in dissociated fetal rat cerebral cortex in vitro. Dev Brain Res 1: 591–605Google Scholar
  22. Shtark MB, Stratievsky VI, Ratushnjak AS, Voskresenskaja LV, Karasev NP (1976) A comparative statistical study of hippocampal neuronal spontaneous spike activity in situ and in vitro. J Neurobiol 7: 551–566Google Scholar
  23. Spitzer NC (1982) The development of electrical excitability. In: Sears TA (ed) Neuronal-glial cell interrelationships. Springer, New York, pp 77–91Google Scholar
  24. SPSSX (1983) User's guide. A complete guide to SPSSX language and operations. McGraw Hill, New YorkGoogle Scholar
  25. Stafstrom CE, Johnston D, Wehner JM, Sheppard JR (1980) Spontaneous neural activity in fetal brain reaggregate cultures. Neuroscience 5: 1681–1689Google Scholar
  26. Van Dongen AMJ, Bretschneider F (1984) Functioning of catfish electroreceptors: bursting discharge pattern of kryptopterus electroreceptors elicited by microelectrode impalement. Comp Biochem Physiol 77A: 647–650Google Scholar
  27. Van Huizen F, Romijn HJ (1985) An improved EPTA staining method for synapses in rat cerebral cortex cultures. J Neurosci Meth 14: 267–271Google Scholar
  28. 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: 67–80Google Scholar
  29. Van Leeuwen FW (1981) An introduction to the immunochemical localization of neuropeptides and neurotransmitters. Acta Histochem Suppl. Band XXIV: S49–77Google Scholar
  30. Wu J-Y (1976) Purification, characterization and kinetic studies of GAD and GABA-T from mouse brain. In: Roberts E et al. (eds) GABA in nervous system function. Raven Press, New York, pp 7–55Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • A. M. M. C. Habets
    • 1
  • A. M. J. Van Dongen
    • 1
  • F. Van Huizen
    • 1
  • M. A. Corner
    • 1
  1. 1.Netherlands Institute for Brain Research, I.W.O.AZ AmsterdamThe Netherlands

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