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Morphological characteristics of cultured olfactory bulb cells

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Abstract

Cultured olfactory bulb cells from embryonic mice had ultrastructural characteristics similar to those of many cell types in the intact adult mouse olfactory bulb. Identified cultured cells included mitral/tufted cells, granule cells, short-axon cells, and fibrous and protoplasmic astrocytes. Cultured neurons were found as individual cells, clusters or aggregates. Clusters consisted of a loose array of neurons that appeared to be densely interconnected by neurites. However, few neurites or fascicles emanated from clusters to adjoining areas. Aggregates consisted of many small, usually rounded, neurons piled on top of one larger neuron, or on more than one, with typically many neurites and fascicles projecting to adjacent aggregates, clusters or individual neurons. Neurites of cultured olfactory bulb cells were well developed, and some were several millimeters long. Synapses were very prominent in these cultures, especially in aggregates, clusters, and fascicles. Electron-lucent, dense-core, and coated vesicles were present. Polarity, shape, and length of the long axis (size) of 815 cultured neurons, identified by positive anti-microtubule-associated protein 2 staining, were documented. Cultured neurons varied in size from 9 to 27 μm, with an average size of 16 μm. Elliptical bipolar (35%), triangular multipolar (21%), and round unipolar (15%) were the most common polarity/shape combinations found in culture. Multipolar, triangular, triangular multipolar, and elliptical bipolar cells increased in size with increasing age of culture. The relative proportions of triangular, multipolar, elliptical multipolar, and triangular multipolar cells decreased, whereas the relative proportions of round, unipolar, and round unipolar cells increased with increasing age of culture. These changes in population subtypes and cell size may indicate continued differentiation and maturation of cultured neurons.

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References

  • Allerand CD (1971) Patterns of neuronal differentiation in developing cultures of neonatal mouse cerebellum: a living and silver impregnation study. J Comp Neurol 142:167–204

    Google Scholar 

  • Burry RW (1983) Antimitotic drugs that enhance neuronal survival in olfactory bulb cell cultures. Brain Res 261:261–175

    Google Scholar 

  • Crandall JE, Jacobson M, Kosik KS (1986) Ontogenesis of microtubule-associate protein 2 (MAP2) in embryonic mouse cortex. Dev Brain Res 28:127–133

    Google Scholar 

  • Currie DN, Dutton GR (1980) [3H] GABA uptake as a marker for cell type in primary cultures of cerebellum and olfactory bulb. Brain Res 199:473–481

    Google Scholar 

  • Dawes CJ (1984) Biological techniques for transmission and scanning electron microscopy, 3rd edn. Ladd Research Industries, Burlington, Vt

    Google Scholar 

  • Denis-Donini S (1989) Expression of dopaminergic phenotypes in the mouse olfactory bulb induced by the calcitonin gene-related peptide. Nature 339:701–703

    Google Scholar 

  • Droge MH, Gross GW, Hightower MH, Czisny LE (1986) Multielectrode analysis of coordinated, multisite, rhythmic bursting in cultured CNS monolayer networks. J Neurosci 6:1583–1592

    Google Scholar 

  • Escobar MI, Pimienta H, Caviness VS Jr, Jacobson M, Crandall JE, Kosik KS (1986) Architecture of apical dendrites in the murine neocortex: dual apical dendritic systems. Neuroscience 17:975–989

    Article  CAS  PubMed  Google Scholar 

  • Fedoroff S (1987) Prenatal ontogenesis of astrocytes. In: Fedoroff S, Vernadakis A (eds) Astrocytes. Development, morphology, and regional specialization of astrocytes, vol 1. Academic Press, New York, 35–74

    Google Scholar 

  • Fedoroff S, Hertz L (eds) (1977) Cell, tissue, and organ cultures in neurobiology. Academic Press, New York

    Google Scholar 

  • Frosch MP, Dichter MA (1984) Physiology and pharmacology of olfactory bulb neurons in dissociated cell culture. Brain Res 290:321–332

    Google Scholar 

  • Gall CM, Hendry SHC, Seroogy KB, Jones EG, Haycock JW (1987) Evidence for coexistence of GABA and dopamine in neurons of the rat olfactory bulb. J Comp Neurol 266:307–318

    Google Scholar 

  • Garber BB (1977) Cell aggregation and recognition in the self-assembly of brain tissues. In: Fedoroff S, Hertz L (eds) Cell, tissue, and organ cultures in neurobiology. Academic Press, New York, pp 515–537

    Google Scholar 

  • Greer CA (1987) Golgi analysis of dendritic organization among denervated olfactory bulb granule cells. J Comp Neurol 257:442–452

    Google Scholar 

  • Gregory GE (1980) Alcoholic Bouin fixation of insect nervous systems for Bodian silver staining. II. Modified solutions. Stain Technol 55:151–160

    Google Scholar 

  • Guo L, Thompson RM, Fracek SP Jr, Schafer R (1989) Dissociated mouse olfactory bulb cell cultures. Soc Neurosci Abstr 15:750

    Google Scholar 

  • Guo L, Thomas T, Schafer R, Fracek SP Jr (1990) Immunocytochemistry of dissociated mouse olfactory bulb cell cultures. Soc Neurosci Abstr 16:403

    Google Scholar 

  • Halasz N, Ljungdahl A, Hokflet T, Johannsson O, Goldstein M, Park D, Biberfeld P (1977) Transmitter histochemistry of the rat olfactory bulb. I. Immunohistochemical localization of monoamine-synthesizing enzymes. Support for intrabulbar, periglomerular dopamine neurons. Brain Res 126:455–474

    Google Scholar 

  • Hatten ME, Liem RKH (1981) Astroglial cells provide a template for the positioning of developing cerebellar neurons in vitro. J Cell Biol 90:622–630

    Google Scholar 

  • Hatten ME, Lynch M, Rydel RE, Sanchez J, Joseph-Silverstein J, Moscatelli D, Rifkd DB (1988) In vitro neurite extension by granule neurons is dependent upon astroglial-derived fibroblast growth factor. Dev Biol 125:280–289

    Google Scholar 

  • Hinds JW (1972a) Early neuron differentiation in the mouse olfactory bulb. I. Light microscopy. J Comp Neurol 146:233–252

    Google Scholar 

  • Hinds JW (1972b) Early neuron differentiation in the mouse olfactory bulb. II. Electron microscopy. J Comp Neurol 146:253–276

    Google Scholar 

  • Hinds JW, Hinds PL (1976a) Synapse formation in the mouse olfactory bulb. I. Quantitative studies. J Comp Neurol 169:15–62

    Google Scholar 

  • Hinds JW, Hinds PL (1976b) Synapse formation in the mouse olfactory bulb. II. Morphogenesis. J Comp Neurol 169:41–62

    Google Scholar 

  • Hinds JW, Ruffett TL (1973) Mitral cell development in the mouse olfactory bulb: reorientation of the perikaryon and maturation of the axon initial segment. J Comp Neurol 151:281–306

    Google Scholar 

  • Kaplan MS, McNelly NA, Hinds JW (1985) Population dynamics of adult-formed granule neurons of the rat olfactory bulb. J Comp Neurol 239:117–125

    Google Scholar 

  • Kosaka K, Hama K, Nagatsu I, Wu J-Y, Ottersen OP, StormMathisen J, Kosaka T (1987) Postnatal development of neurons containing both catecholaminergic and GABAergic traits in the rat main olfactory bulb. Brain Res 403:355–360

    Google Scholar 

  • Kosik KS, Finch EA (1987) MAP2 and tau segregate into dendritic and axonal domains after the elaboration of morphologically distinct neurites: an immunocytochemical study of cultured rat cerebellum. J Neurosci 7:3142–3153

    Google Scholar 

  • Levitt P, Moore RY, Garber B (1976) Selective cell association of catecholamine neurons in brain aggregates in vitro. Brain Res 111:311–320

    Google Scholar 

  • Loots GP, Loots JM, Brown JMM, Schoeman JL (1979) A rapid silver impregnation method for nervous tissue: a modified protargol-peroxide technic. Stain Technol 54:97–100

    Google Scholar 

  • Lucas JH, Czisny LE, Gross GW (1986) Adhesion of cultured mammalian central nervous system neurons to flame-modified hydrophobic surfaces. In Vitro Cell Dev Biol 22:37–413

    Google Scholar 

  • Macrides F, Schneider SP (1982) Laminar organization of mitral and tufted cells in the main olfactory bulb of the adult hamster. J Comp Neurol 208:419–430

    Google Scholar 

  • McCarthy KD, Vellis J de (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85:890–902

    Google Scholar 

  • McLean JH, Shipley MT (1988) Postmitotic, postmigrational expression of tyrosine hydroxylase in olfactory bulb dopaminergic neurons. J Neurosci 8:3658–3669

    Google Scholar 

  • Mori K, Kishi K, Ojima H (1983) Distribution of dendrites of mitral, displaced mitral, tufted, and granule cells in the rabbit olfactory bulb. J Comp Neurol 219:339–355

    Google Scholar 

  • Neale EA, Moonen G, Macdonald RL, Nelson PG (1982) Cerebellar macroneurons in microexplant cell culture: ultrastructural morphology. Neuroscience 7:1879–1890

    Google Scholar 

  • O'Lague PH, Potter DD, Furshpan EJ (1978) Studies on rat sympathetic neurons developing in cell culture. I. Growth characteristics and electrophysiological properties. Dev Biol 67:384–403

    Google Scholar 

  • Peacock JH, Rush DF, Mathers LH (1979) Morphology of dissociated hippocampal cultures from fetal mice. Brain Res 169:231–246

    Google Scholar 

  • Peters A, Palay SL, Webster HdeF (1991) The fine structure of the nervous system: neurons and their supporting cells, 3rd edn. Oxford University Press, New York

    Google Scholar 

  • Pinching AJ, Powell TPS (1971) The neuropil of the glomeruli of the olfactory bulb. J Cell Sci 9:347–377

    CAS  PubMed  Google Scholar 

  • Price JL, Powell TPS (1970a) The mitral and short axon cells of the olfactory bulb. J Cell Sci 7:631–651

    Google Scholar 

  • Price JL, Powell TPS (1970b) The morphology of the granule cells of the olfactory bulb. J Cell Sci 7:91–123

    Google Scholar 

  • Price JL, Powell TPS (1970c) The synaptology of the granule cells of the olfactory bulb. J Cell Sci 7:125–155

    Google Scholar 

  • Ramakers GJA, Corner MA, Habets AMMC (1990) 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

    Google Scholar 

  • Ransom BR, Christian CN, Bullock PN, Nelson PG (1977a) Mouse spinal cord in cell culture. II. Synaptic activity and circuit behavior. J Neurophysiol 40:1151–1162

    Google Scholar 

  • Ransom BR, Neale E, Henkart M, Bullock PN, Nelson PG (1977b) Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties. J Neurophysiol 40:1132–1150

    Google Scholar 

  • Sensenbrenner M, Mandel P (1974) Behaviour of neuroblasts in the presence of glial cells, fibroblasts and meningeal cells in culture. Exp Cell Res 87:159–167

    Google Scholar 

  • Shepherd GM, Greer CA (1990) Olfactory bulb. In: Shepherd GM (ed) The synaptic organization of the Brain, 3rd edn. Oxford University Press, New York, 111–258

    Google Scholar 

  • Trombley PQ, Westbrook GL (1990) Excitatory synaptic transmission in cultures of rat olfactory bulb. J Neurophysiol 64:598–606

    Google Scholar 

  • Willey TJ (1973) The ultrastructure of the cat olfactory bulb. J Comp Neurol 152:211–232

    Google Scholar 

  • Zar JH (1984) Biostatistical analysis. Prentice-Hall, Englewood Cliffs, NJ

    Google Scholar 

Download references

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Author notes

  1. S. P. Fracek Jr.

    Present address: Department of Humanities and Basic Sciences, The University of Texas Medical Branch at Galveston, School of Allied Health Sciences, 11th and Mechanic Sts., 77555-1028, Galveston, TX, USA

Authors and Affiliations

  1. Department of Biological Sciences and Center for Network Neuroscience, University of North Texas, Box 5218, 76203-5218, Denton, TX, USA

    S. P. Fracek Jr., L. Guo & R. Schafer

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  1. S. P. Fracek Jr.
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  2. L. Guo
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  3. R. Schafer
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Fracek, S.P., Guo, L. & Schafer, R. Morphological characteristics of cultured olfactory bulb cells. Exp Brain Res 79, 421–436 (1994). https://doi.org/10.1007/BF00229182

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  • DOI: https://doi.org/10.1007/BF00229182

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