Abstract
The principles of oriented growth of nerve fibers and the formation of functional synaptic connections during combined culture of brain structures with no direct anatomical or functional connections (the spinal cord and olfactory bulbs of mouse embryos) were investigated by neuromorphological and electrophysiological methods. During the first week of culture connections, mainly glio-neuronal bridges, formed between the explants of spinal cord and olfactory bulb. Glial cells forming an oriented substrate, facilitating growth and fasciculation (the formation of bundles) of axons that developed subsequently, play an active role in the formation of such connections. In preparations impregnated with silver, connections formed by bundles of axons or by single nerve fibers were seen between the explants. The results of electrophysiological investigations of combined cultures of heterogeneous brain structures showed that by the second week of culture functional synaptic connections have formed between explants of spinal cord and olfactory bulbs. Electrical stimulation of spinal cord explants led to the appearance of short- and long-latency unit responses in explants of the olfactory bulbs. The formation of nonspecific functional synaptic connections for these brain structures during combined culture, revealed by this investigation, is evidence of the high level of morphogenetic plasticity of growing or regenerating axons and of the active role of neuroglial cells in preparation and provision for oriented growth of nerve fibers.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature cited
I. V. Viktorov, "Culture by Maximow's method," in: Textbook of Nerve Tissue Culture [in Russian], Nauka, Moscow (1976), pp. 21–28.
I. V. Viktorov, "Formation of connections between isolated organotypic brain explants," in: Functional and Structural Bases of Activity of Brain Systems and Mechanisms of Brain Plasticity [in Russian], Brain Institute, Academy of Medical Sciences of the USSR, Moscow (1976), pp. 78–82.
S. H. Appel, "Neuronal recognition and synaptogenesis," Exp. Neurol.,48, 52 (1975).
M. E. Bernstein and J. J. Bernstein, "Regeneration of axons and synaptic complex formation rostral to the site of hemisection in the spinal cord of the monkey," Int. J. Neurosci.,5, 15 (1973).
R. P. Bunge, "The expression of neuronal specificity in tissue culture," in: Neuronal Recognition, S. H. Barondes, ed., London, Chapman and Hall (1976), pp. 109–127.
J. H. Chamley and J. J. Dowell, "Specificity of nerve fiber attraction to autonomic effector organs in tissue culture," Exp. Cell Res.,90, 1 (1975).
C. W. Cotman and G. S. Lynch, "Reactive synaptogenesis in the adult nervous system. The effect of partial deafferentation on new synapse formation," in: Neuronal Recognition, edited by S. H. Barondes, London, Chapman and Hall (1976), pp. 69–108.
S. M. Crain, Neurophysiologic Studies in Tissue Culture, Raven Press, New York (1976).
S. M. Crain and E. R. Peterson, "Development of specific sensory-evoked synaptic networks in fetal mouse cord-brainstem cultures," Science, 188, 275 (1975).
T. Ebendall and C. O. Jacobsen, "Tissue explants affecting extension and orientation of axons in culture chick embryo ganglia," Exp. Cell Res.,105, 379 (1977).
D. W. James and R. L. Tressman, "Synaptic profiles in the outgrowth from chick spinal cord in vitro," Z. Zellforsch,101, 598 (1969).
J. Raisman, "Neuronal plasticity in the septal nuclei of the adult rat," Brain Res.,14, 25 (1969).
R. R. Ruffolo, G. Eisenbarth, J. M. Thompson, and M. Nirenberg, "Synapse turnover. A mechanism for acquiring synaptic specificity," Proc. Natl. Acad. Sci. USA,78, 2281 (1978).
G. R. Schneider, "Early lesions of superior colliculus. Factors affecting the formation of abnormal retinal projection," Brain Behav. Evol.,8, 73 (1973).
R. L. Sidman and N. K. Wessels, "Control of direction of growth during the elongation of neurites," Exp. Neurol.,48, 237 (1975).
M. Singer, R. H. Norlander, and M. Egar, "Axonal guidance during embryogenesis in the spinal cord of the newt: the blueprint hypothesis of neuronal pathway patterning," J. Comp. Neurol.,185, 1 (1979).
N. R. Smalheiser and S. M. Crain, "Formation of functional retinotectal connections in co-cultures of fetal mouse explants," Brain Res.,148, 484 (1978).
H. M. Sobkowicz, R. M. Guillery, and M. B. Bornstein, "Neuronal organization in long term cultures of the spinal cord of the fetal mouse," J. Comp. Neurol.,132, 365 (1968).
R. W. Sperry, "Chemoaffinity in the orderly growth of nerve fiber patterns and connections," Proc. Natl. Acad. Sci. USA,50, 703 (1963).
W. R. Stevens, D. W. Slaaf, J. Hooisma, T. Magchielse, and E. Meeter, "Neurotransmission and specificity of innervation in mixed culture of embryonic ciliary ganglia and skeletal muscle cells," in: Maturation of the Nervous System, edited by M. A. Corner, Amsterdam, Elsevier, pp. 21–29 (Progr. Brain Res., Vol. 48).
S. Varon, "Neural growth and regeneration. A cellular perspective," Exp. Neurol.,54, 1 (1977).
P. Weiss, "Nervous systems," in: Analysis of Development, edited by B. H. Willier, P. Weiss, and V. Hamburger. Philadelphia: Saunders (1955), pp. 346–401.
Additional information
Brain Institute, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 490–497, September–October, 1980.
Rights and permissions
About this article
Cite this article
Viktorov, I.V., Sharonova, I.N. Formation of functional synaptic connections between heterogeneous brain formations in organotypic nerve tissue culture. Neurophysiology 12, 311–317 (1980). https://doi.org/10.1007/BF01066077
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01066077