Summary
(Gamma aminobutyric acid) GABA was applied to cultures of mouse neuroblastoma cells of different ages at concentrations ranging from 10-4 to 10-6 M. The cultures were exposed to GABA either in short term experiments for 2 h to 2 days or for longer periods by adding the substance twice within 10 days at 5-day intervals. The following effects were observed: (1) There was a strong proliferation of coated vesicles, appearing to derive from the Golgi complex and the rough endoplasmic reticulum (RER), and also showing all intermediate stages of fusion and pinching off from the plasma membranes. (2) In numerous areas, electron-dense material aggregated at the inner aspect of the plasma membrane and around small invaginations of the plasmalemma. (3) The number and area of specialized contacts increased between cells and their processes. (4) Similar to cultures free of GABA, varicosities and terminal swellings of the cells and their processes were filled with small round vesicles, 40–60 nm in diameter, or with smooth, very large, empty-appearing vesicular inclusions, or with flat pleiomorphic vesicles. In addition, mitochondria and some formations of the smooth endoplasmic reticulum (SER) appeared, and primitive contacts (symmetrical densities) were formed. (5) Dense-cored vesicles were found peripherally and linearly arranged, surrounded by an electron-dense substance. (6) Electron-dense material of unknown origin was seen between cells or their processes near the peripherally arranged dense-cored vesicles. Exogenous GABA may play a specific role in the early stages of synaptogenesis, since it showed a positive effect on the neuroblastoma cells, which in the absence of GABA are only capable of forming primitive or immature presynaptic elements. The significance of the peripheral accumulation of dense-cored vesicles, accompanied by an amorphous, electron-dense substance occurring both intra- and extracellularly is discussed.
Similar content being viewed by others
Reference
Adams PR, Brown DA (1975) Actions of γ-aminobutyric acid on sympathetic ganglion cells. J Physiol (London) 250:85–120
Altman J (1971) Coated vesicles and synaptogenesis. A developmental study in the cerebellar cortex of the rat. Brain Res 34:311–322
Breakefield XO, Neale EA, Neale JH, Jacobowitz DM (1975) Localized catecholamine storage associated with granules in murine neuroblastoma cells. Brain Res 92:237–256
Bunge MB (1973) Fine stucture of nerve fibers and growth cones of isolated sympathetic neurons in culture. J Cell Biol 56:713–735
Bunge RP, Bunge MB, Peterson ER (1965) An electron microscope study of cultured rat spinal cord. J Cell Biol 24:163–191
Bunge MB, Bunge RP, Peterson ER (1967) The onset of synapse formation in spinal cord cultures as studied by electron microscopy. Brain Res 6:728–749
Burry RW (1980a) Formation of apparent presynaptic elements in response to poly-basic compounds. Brain Res 184:85–98
Burry RW (1980b) Dense material associated with apparent presynaptic elements in cell cultures of the CNS. Cell Tissue Res 211:95–103
Chan KY, Bunt AH, Haschke RH (1980) Endocytosis and compartmentation of peroxidases and cationized ferritin in neuroblastoma cell. J Neurocytol 9:381–403
Crain SM (1966) Development of organotypic bioelectric activities in central nervous tissues during maturation culture. In: Pfaffer CC, Smythies JR (eds) Intl Rev Neurobiol, Academic Press, New York, Vol 9, pp 1–43
Glees P, Sheppard BL (1964) Electron microscopical studies of the synapse in the developing chick spinal cord. Z Zellforsch 62:356–362
Hinds JW, Hinds PL (1976) Synapse formation in the mouse olfactory bulb. II Morphogenesis. J Comp Neurol 169:41–62
Joó F, Dames W, Wolff JR (1979) Effect of prolonged sodium bromide administration on the fine structure of dendrites in the superior cervical ganglion of adult rat. Progress in Brain Research 51:109–115
Kanaseki T, Kadota K (1969) The “vesicle in a basket”. A morphological study of the coated vesicle isolated from the nerve endings of the guinea pig brain with special reference to the mechanism of membrane movements. J Cell Biol 42:202–220
Klebe R (1970) Genetic papping of a human regulator gene. Ph D dissertation. Dept Biol Yale Univ
Klebe R, Ruddle F (1969) Neuroblastoma: Cell culture analysis of a differentiating stem cell system. J Cell Biol 43:69A
Krstić RV (1976) Ultrastruktur der Säugetierzelle. Springer-Verlag, Berlin Heidelberg New York, pp 60–69
Pappas GD, Peterson ER, Masurorsky ER, Crain SM (1971) Electron microscopy of the in vitro development of mammalian motor end plates. Ann NY Acad Sc 183:33–45
Pinching AJ (1969) Persistance of post-synaptic thickenings after degeneration of olfactory nerves. Brain Res 16:277–281
Raisman G, Field PM, Ostberg AJC, Iversen LL, Zigmond RE (1974) A quantitative ultrastructural and biochemical analysis of the process of reinervation of the superior cervical ganglion in the adult rat. Brain Res 71:1–16
Rees RP, Bunge MB, Bunge RP (1976) Morphological changes in the neuritic growth cone and target neuron during synaptic junction development in culture. J Cell Biol 68:240–263
Ross J, Olmsted BJ, Rosenbaum JL (1975) The ultrastructure of mouse neuroblastoma cells in tissue culture. Tissue Cell 7:107–136
Spoerri PE, Dresp W (1981) Smooth endoplasmic reticulum and synaptic vesicles in cultured murine neuroblastoma cells. Z mikrosk-anat Forsch 95: Heft 5 (in press)
Spoerri PE, Dresp W, Heyder E (1980a) A simple embedding technique for monolayer neuronal cultures grown in plastic flasks Acta Anat 107:221–223
Spoerri PE, Glees P, Dresp W (1980b) The time course of synapse formation of mouse neuroblastoma cells in monolayer cultures. Cell Tissue Res 205:441–421
Stelzner DJ (1971) The relations between synaptic vesicles, Golgi apparatus and smooth endoplasmic reticulum; a developmental study using the zinc iodide-osmium technique. Z Zellforsch 120:332–345
Stelzner DJ, Martin AH, Scott GL (1973) Early stages of synaptogenesis in the cervical spinal cord of the chick embryo. Z Zellforsch 138:475–488
Ticku MK, Huang A, Barker JL (1980) Characterization of γ-aminobutyric receptor binding in cultured brain cells. Mol Pharmacol 17:285–289
Waxman SG, Pappas GD (1969) Pinocytosis at postsynaptic membranes: electron microscopic evidence. Brain Res 14:240–244
Westrum LE (1975) Electron microscopy of synaptic structures in olfactory cortex of early postnatal rats. J Neurocytol 4:713–732
Wolff JR (1978) Ontogenic aspects of cortical architecture: lamination. In: Brazier MAB, Petsche H (eds) Architectonics of the Cerebral Cortex. Raven Press, New York, pp 159–173
Wolff JR (1979) Hinweise auf eine Doppelrolle von GABA als synaptischer Transmitter und als morphogenetischer Faktor In: Verh Dtsch Zool Ges. Gustav Fischer Verlag Stuttgart 1979, 194–200
Wolff JR, Joó F, Dames W (1978) Plasticity in dendrites shown by continuous GABA administration in superior cervical ganglion of adult rat. Nature 274:72–74
Wolff JR, Rickmann M, Chronwall BM (1979a) Axo-glial synapses and GABA-accumulating glial cells in the embryonic neocortex of the rat. Cell Tissue Res 201:239–248
Wolff JR, Joó F, Dames W, Fehér O (1979b) Induction and maintainance of free postsynaptic thickenings in the adult superior cervical ganglion. J. Neurocytol 8:849–863
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Spoerri, P.E., Wolff, J.R. Effect of GABA-Administration on murine neuroblastoma cells in culture. Cell Tissue Res. 218, 567–579 (1981). https://doi.org/10.1007/BF00210116
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00210116