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Influence of meningeal cells on the proliferation and maturation of rat neuroblasts in culture

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Summary

Neuronal cells were obtained by dissociating cells from the cerebral hemispheres of rat embryos (10 to 17-day-old), either cleaned entirely or only partially of their meningeal membranes. These cells were seeded on poly-lysine-coated Petri dishes in serum-containing medium. The cultures most enriched in neuronal cells were obtained from brains of 13- to 15-day-old embryos and after 2 h, the culture medium was switched to Dulbecco's modified Eagle's medium, without serum, supplemented with the N1 supplements as described by Bottenstein et al. (1980). The proliferation of neuroblasts from 13-day-old embryos in the presence or absence of meningeal cells was studied by using a combination of tritiated thymidine autoradiography and immuno-staining against neurofilament proteins. The neuroblasts seem to proliferate during the first 3 days. The proliferative activity was further enhanced in the presence of meningeal cells. The glioblasts multiply only after a period of one week in culture conditions as observed here. The subsequent development of the neuroblasts was followed over a period of 4 weeks and the ultrastructural appearance of these cells was investigated at 2 and 3 weeks. In the presence of meningeal cells, many neurons, intensely stained for neurofilament proteins, survived for 21 days, while in control cultures they underwent massive degeneration after 2 weeks. Synapses with numerous clear vesicles were abundant in cultures grown under the influence of meningeal cells; they were rare and possessed few vesicles in control cultures. The data indicate that meningeal cells affect the proliferation and maturation of rat neuroblasts in culture.

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References

  • Ahmed Z, Walker PS, Fellows RE (1983) Properties of neurons from dissociated fetal rat brain in serum-free culture. J Neurosci 3: 2448–2462

    Google Scholar 

  • Allen C, Sievers J, Berry M, Jenner S (1981) Experimental studies on cerebellar foliation. II. A morphometric analysis of cerebellar fissuration defects and growth retardation after neonatal treatment with 6-OHDA in the rat. J Comp Neurol 203: 771–783

    Google Scholar 

  • Asou H, Iwasaki N, Hirano S, Dahl D (1985) Mitotic neuroblasts in dissociated cell cultures from embryonic rat cerebral hemispheres express neurofilament protein. Brain Res 332: 355–357

    Google Scholar 

  • Banker GA, Cowan WM (1977) Rat hippocampal neurons in dispersed cell culture. Brain Res 126: 397–425

    Google Scholar 

  • Barakat I, Sensenbrenner M, Vincendon G (1982) The importance of cell contact for the proliferation of neuroblasts in culture and its stimulation by meningeal extract. Neurochem Res 7: 287–300

    Google Scholar 

  • Barakat I, Wittendorp-Rechenmann E, Rechenmann RV, Sensenbrenner M (1981) Influence of meningeal cells on the proliferation of neuroblasts in culture. Dev Neurosci 4: 363–372

    Google Scholar 

  • Barde Y-A, Edgar D, Thoenen H (1983) New neurotrophic factors. Ann Rev Physiol 45: 601–612

    Google Scholar 

  • Baudysova M, Michl J (1982) Serum-free medium is more beneficial than fetal calf serum-supplemented medium for morphological differentiation of primary chick embryo brain cells in culture. Mol Physiol 2: 225–233

    Google Scholar 

  • Berg DK (1984) New neuronal growth factors. Ann Rev Neurosci 7: 149–170

    Google Scholar 

  • Berger B, Di Porzio U, Daguet MC, Gay M, Vigny A, Glowinski J, Prochiantz A (1982) Long-term development of mesencephalic dopaminergic neurons of mouse embryos in dissociated primary cultures: morphological and histochemical characteristics. Neuroscience 7: 193–205

    Google Scholar 

  • Bird MM (1983) Neurons and glial cells in long term cultures of previously dissociated newborn mouse cerebral cortex. J Anat 136: 293–305

    Google Scholar 

  • Borg J, Spitz B, Hamel G, Mark J (1985) Selective culture of neurons from rat cerebral cortex: morphological characterization, glutamate uptake and related enzymes during maturation in various culture media. Dev Brain Res 18: 37–49

    Google Scholar 

  • Bottenstein JE, Skaper SD, Varon SS, Sato GH (1980) Selective survival of neurons from chick embryo sensory ganglionic dissociates utilizing serum-free supplemented medium. Exp Cell Res 125: 183–190

    Google Scholar 

  • Cam Y, Sensenbrenner M, Mandel P (1975) A comparative study of the effects of brain extracts and mesodermal membrane extracts on nerve cell differentiation. Experientia 31: 1430–1431

    Google Scholar 

  • Cochard P, Paulin D (1984) Initial expression of neurofilaments and vimentin in the central and peripheral nervous system of the mouse embryo in vivo. J Neurosci 4: 2080–2094

    Google Scholar 

  • Debbage P (1985) Distinctive structural and cytoskeletal properties of the long-surviving neurons in cell cultures of embryonic spinal cord. Neuroscience 14: 207–224

    Google Scholar 

  • Delacourte A, Filliatreau G, Boutteau F, Bisente G, Schrevel J (1980) A study of the 10 mm filament fraction isolated during the standard microtubule preparation. Biochem J 191: 543–546

    Google Scholar 

  • Faivre-Bauman A, Rosenbaum E, Puymirat J, Grouselle D, Tixier-Vidal A (1981) Differentiation of fetal mouse hypothalamic cells in serum-free medium. Dev Neurosci 4: 118–129

    Google Scholar 

  • Fedoroff S, Krukoff TL, Fisher KRS (1982) The development of chick spinal cord in tissue culture. III. Neuronal precursor cells in culture. In Vitro 18: 183–195

    Google Scholar 

  • Juurlink BHJ, Fedoroff S (1982) The development of mouse spinal cord in tissue culture. II. Development of neuronal precursor cells. In Vitro 18: 179–182

    Google Scholar 

  • Keller M, Jackisch R, Seregi A, Hertting G (1985) Comparison of prostanoid forming capacity of neuronal and astroglial cells in primary cultures. Neurochem Int 7: 655–665

    Google Scholar 

  • Kingsbury AE, Gallo V, Woodhams PL, Balazs R (1985) Survival, morphology and adhesion properties of cerebellar interneurones cultured in chemically defined and serum-supplemented medium. Dev Brain Res 17: 17–25

    Google Scholar 

  • Korr H (1980) Proliferation of different cell types in the brain. In: Advances in anatomy embryology and cell biology. Springer, Berlin, Vol 61, pp 1–71

    Google Scholar 

  • Korr H (1981) Light microscopical autoradiography of nervous tissue. In: Heyn Ch, Forssmann WG (eds). Techniques in neuroanatomical research. Springer, Berlin, pp 218–244

    Google Scholar 

  • Kriegstein A, Dichter MA (1984) Neuron generation in dissociated cell cultures from fetal cerebral cortex. Brain Res 295: 184–189

    Google Scholar 

  • Kumar S (1983) Growth of neural cell cultures in a chemically defined, serum-free culture medium. Neurochem Res 8: 847–852

    Google Scholar 

  • Kurihara T, Tsukada Y (1967) The regional and subcellular distribution of 2′,3′-cyclic nucleotide 3′-phosphohydrolase in the central nervous system. J Neurochem 14: 1167–1174

    Google Scholar 

  • Landesman R (1967) Neural-mesodermal interactions subsequent to neural induction in Ambystoma. Devl Biol 16: 341–367

    Google Scholar 

  • Leterrier J-F, Wong J, Liem RKH, Shelanski ML (1984) Promotion of microtubule assembly by neurofilament-associated microtubule-associated proteins. J Neurochem 43: 1385–1391

    Google Scholar 

  • Löffler F, Lohmann SM, Walckhoff B, Walter U, Hamprecht B. (1986) Immunocytochemical characterization of neuron-rich primary cultures of embryonic rat brain cells by established neuronal and glial markers and by monospecific antisera against cyclic nucleotide-dependent protein kinases and the synaptic vesicle protein synapsin I. Brain Res 363: 205–221

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275

    CAS  PubMed  Google Scholar 

  • Messer A (1981) Primary monolayer cultures of the rat corpus striatum: morphology and properties related to acetylcholine and γ-aminobutyrate. Neuroscience 6: 2677–2687

    Google Scholar 

  • Pehlemann FW, Sievers J, Berry M (1985) Meningeal cells are involved in foliation, lamination, and neurogenesis of the cerebellum: evidence from 6-hydroxydopamine-induced destruction of meningeal cells. Dev Biol 110: 136–146

    Google Scholar 

  • Pettmann B, Louis JC, Sensenbrenner M (1979) Morphological and biochemical maturation of neurones cultured in the absence of glial cells. Nature 281: 378–380

    Google Scholar 

  • Raju T, Bignami A, Dahl D (1981) In vivo and in vitro differentiation of neurons and astrocytes in the rat embryo. Dev Biol 85: 344–357

    Google Scholar 

  • Romijn HJ, Habets AMMC, Mud MT, Wolters PS (1982) Nerve outgrowth, synaptogenesis and bioelectric activity in fetal rat cerebral cortex tissue cultured in serum-free, chemically defined medium. Dev Brain Res 2: 583–589

    Google Scholar 

  • Romijn HJ, Van Huizen F, Wolters PS (1984) Towards an improved serum-free, chemically defined medium for long-term culturing of cerebral cortex tissue. Neurosci Biobehav Rev 8: 301–334

    Google Scholar 

  • Ruegg UT, Hefti F (1984) Growth of dissociated neurons in culture dishes coated with synthetic polymeric amines. Neurosci Lett 49: 319–324

    Google Scholar 

  • Sensenbrenner M, Devilliers G, Bock E, Porte A (1980) Biochemical and ultrastructural studies of cultured rat astroglial cells. Effect of brain extract and dibutyryl cyclic AMP on glial fibrillary acidic protein and glial filaments. Differentiation 17: 51–61

    Google Scholar 

  • Sensenbrenner M, Maderspach K, Latzkovits L, Jaros GG (1978) Neuronal cells from chick embryo cerebral hemispheres cultivated on polylysine-coated surfaces. Dev Neurosci 1: 90–101

    Google Scholar 

  • Sensenbrenner M, Wittendorp E, Barakat I, Rechenmann RV (1980) Autoradiography study of proliferating brain cells in culture. Dev Biol 75: 268–277

    Google Scholar 

  • Skaper SD, Adler R, Varon S (1979) A procedure for purifying neuron-like cells in cultures from central nervous tissue with a defined medium. Dev Neurosci 2: 233–237

    Google Scholar 

  • Sotelo J, Gibbs CJ Jr, Gajdusek C, Toh BH, Wurth M (1980) Method for preparing cultures of central neurons: cytochemical and immunochemical studies. Proc Natl Acad Sci USA 77: 653–657

    Google Scholar 

  • Thampy KG, Sauls CD, Brinkley BR, Barnes EM Jr (1983) Neurons from chick embryo cerebrum: ultrastructural and biochemical development in vitro. Dev Brain Res 8: 101–110

    Google Scholar 

  • Toivenen S, Saxen L (1968) Morphogenetic interaction of presumptive neural and mesodermal cells mixed in different ratio. Science 159: 539–540

    Google Scholar 

  • Weyhenmeyer J, Bright MJ (1983) Expression of neuron-specific enolase in cultured neurons from the fetal rat. Neurosci Lett 43: 303–307

    Google Scholar 

  • Yavin E, Yavin Z (1974) Attachment and culture of dissociated cells from rat embryo cerebral hemispheres on polylysinecoated surface. J Cell Biol 62: 540–546

    Google Scholar 

  • Yavin Z, Yavin E (1980) Survival and maturation of cerebral neurons on poly(L-lysine) surfaces in the abscence of serum. Dev Biol 75: 454–459

    Google Scholar 

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Authors and Affiliations

  1. Centre de Neurochimie du CNRS, 5, Rue Blaise Pascal, F-67084, Strasbourg Cédex, France

    C. Gensburger, G. Labourdette & M. Sensenbrenner

  2. INSERM U.44, 5, Rue Blaise Pascal, F-67084, Strasbourg Cédex, France

    G. Labourdette

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  1. C. Gensburger
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  2. G. Labourdette
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  3. M. Sensenbrenner
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Gensburger, C., Labourdette, G. & Sensenbrenner, M. Influence of meningeal cells on the proliferation and maturation of rat neuroblasts in culture. Exp Brain Res 63, 321–330 (1986). https://doi.org/10.1007/BF00236849

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  • DOI: https://doi.org/10.1007/BF00236849

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