Skip to main content
Log in

Unipolar brush cells develop a set of characteristic features in primary cerebellar cultures

  • Published:
Journal of Neurocytology

Abstract

The unipolar brush cells (UBCs) are a class of excitatory interneurons recently discovered in the cerebellar granular layer. UBCs differ morphologically and biochemically from granule cells, although they share the same mossy fiber and Golgi cell inputs. To elucidate development of the UBCs, we sought to ascertain their presence in primary cerebellar cultures and the class-specific properties they develop in vitro outside of the context of the tissue. By light and electron microscopy, we demonstrate that primary cultures from embryonic and postnatal mouse and rat cerebella contain UBC-like neurons that are highly polarized and can be distinguished from granule cells on several grounds. Granule cells are more numerous in dissociated postnatal cultures than in embryonic cultures; express little, if any, calretinin immunoreactivity; and develop dendritic processes devoid of typical claw-like endings, but provided with small synaptic junctions. By contrast, UBC-like neurons occur more frequently in embryonic cultures than in postnatal cultures, are intensely calretinin-positive, and develop characteristic cell organelles, dendrites, and large synapses. In embryonic cultures, the UBC-like neurons have a clear nucleus and contain a special cytoplasmic array of ringlet subunits, resembling the botrysome. At 12–28 days in vitro, the UBC dendrites contain abundant mitochondria, are provided with clusters of non-synaptic appendages, and engage in glomerular arrays together with large and small axon terminals. The large terminals contain round synaptic vesicles, form extensive, asymmetric synapses with the cell bodies and the dendrites of the UBC-like neurons, and resemble mossy terminals, while the small terminals contain pleomorphic vesicles, form symmetric synaptic junctions, and resemble Golgi terminals. In postnatal cultures grown for 12 days, UBC-like neurons are rare and resemble in most aspects the cells observed in embryonic cultures, although they rarely develop elaborate dendritic brushes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • ABBOTT, L. C. & JACOBOWITZ, D. M. (1995) Development and calretinin immunoreactive unipolar brush-like cells and an afferent pathway to the embryonic and early postnatal mouse cerebellum. Anatomy and Embryology 191, 541–559.

    Google Scholar 

  • ALTMAN, J. & BAYER, S. A. (1996) Development of the Cerebellar System. Boca Raton, Florida: CRC Press.

    Google Scholar 

  • ANELLI, R., KETTNER, R. E. & MUGNAINI, E. (1996) Unipolar brush cells are present in cerebellar granule cell in cultures. Society for Neuroscience Abstracts 22, 1631.

    Google Scholar 

  • ANELLI, R. & MUGNAINI, E. (1998) Characterization of cerebellar unipolar brush cells in vitro. Society for Neuroscience Abstracts 24, 2005.

    Google Scholar 

  • ANELLI, R., SAKANE, H. & MUGNAINI, E. (1997) Calretinin positive neurons in dissociated cerebellar cultures. Society for Neuroscience Abstracts 23, 1830.

    Google Scholar 

  • ARAI, R. WINSKY, L. M., ARAI, M. & JACOBOWITZ, D. M. (1991) Immunohistochemical localization of calretinin in the rat hindbrain. Journal of Comparative Neurology 310, 21–44.

    Google Scholar 

  • BAADER, S. L., GRUCHMANN, S. & SCHILLING, K. (1999) The differentiation of cerebellar interneurons is independent of their mitotic history. Neuroscience 90, 1243–1254.

    Google Scholar 

  • BAPTISTA, C. A., HATTEN, M. E., BLAZESKI, R. & MASON, C. A. (1994) Cell-cell interactions influence survival and differentiation of purified Purkinje cells in vitro. Neuron 12, 243–260.

    Google Scholar 

  • BATINI, C., COMPOINT, C., BUISSERET-DELMAS, C., DANIEL, H. & GUEGAN, M. (1992) Cerebellar nuclei and the nucleocortical projections in the rat: Retrograde tracing coupled to GABA and glutamate immunocytochemistry. Journal of Comparative Neurology 315, 74–84.

    Google Scholar 

  • BERTHIÉ, B. & AXELRAD, H. (1994) Granular layer collaterals of the unipolar brush cell axon display rosette-like excrescences. A Golgi study in the rat cerebellar cortex. Neuroscience Letters 167, 161–165.

    Google Scholar 

  • CHAN-PALAY, V. & PALAY, S. L. (1971) The synapse en marron between Golgi type II neurons and mossy fibers in the rat's cerebellar cortex. Zeitschrifts fûr Anatomie und Entwicklungsgeschichte 133, 274–287.

    Google Scholar 

  • DIÑO, M. R., SEKERKOVÁ, G., PERACHIO, A. A. & MUGNAINI, E. (1997) Unipolar brush cells are targets of primary vestibular fibers. Society for Neuroscience Abstracts 23, 1830.

    Google Scholar 

  • DIÑO, M. R., WILLARD, F. H. & MUGNAINI, E. (1999) Distribution of unipolar brush cells and other calretinin immunoreactive components in the mammalian cerebellar cortex. Journal of Neurocytology 28, 99–126.

    Google Scholar 

  • DUNN, M. E., SCHILLING, K. & MUGNAINI, E. (1998a) Development and fine structure of murine Purkinje cells in dissociated cerebellar culture: Neuronal polarity. Anatomy and Embryology 197, 9–29.

    Google Scholar 

  • DUNN, M. E., SCHILLING, K. & MUGNAINI, E. (1998b) Development and fine structure of murine Purkinje cells in dissociated cerebellar culture: Dendritic differentiation, synaptic maturation, and formation of cell-class specific features. Anatomy and Embryology 197, 31–50.

    Google Scholar 

  • DUNN, M. E., VETTER, D. E., BERREBI, A. S., KRIDER, H. M. & MUGNAINI, E. (1996) The mossy fiber-granule cell-cartwheel cell system in the mammalian cochlear nuclear complex. In Advances in Speech, Hearing and Language Processing, Vol 3: Cochlear Nucleus; Structure and Function in Relation to Modelling. (edited by AINSWORTH, W. A.), pp. 63–87. Greenwich, Connecticut: JAI Press.

    Google Scholar 

  • FLORIS, A., DIÑO, M. R., JACOBOWITZ, D. M. & MUGNAINI, E. (1994) The unipolar brush cells of the rat cerebellar cortex and cochlear nucleus are calretininpositive: A study by light and electron microscopic immunocytochemistry. Anatomy and Embryology 189, 495–520.

    Google Scholar 

  • FLORIS, A., DUNN, M. E., BERREBI, A. S., JACOBOWITZ, D. M. & MUGNAINI, E. (1992) Pale cells of the flocculonodular lobe are calretinin positive. Society for Neuroscience Abstracts 18, 853.

    Google Scholar 

  • GARTHWAITE, J. & BALÁZS, R. (1981) Excitatory amino acid-induced changes in cyclic GMP levels in slices and cell suspensions fromcerebellum. Advances in Biochemical Psychopharmacology 27, 317–326.

    Google Scholar 

  • HATTEN, M. E. (1985) Neuronal regulation of astroglial morphology and proliferation in vitro. Journal of Cell Biology 100, 384–396.

    Google Scholar 

  • JAARSMA, D., DIÑO, M. R., COZZARI, C. & MUGNAINI, E. (1996) Cerebellar choline acetyltransferase positive mossy fibres and their granule cells and unipolar brush cell targets: A model for central cholinergic nicotinic neurotransmission. Journal of Neurocytology 25, 829–842.

    Google Scholar 

  • JAARSMA, D., DIÑO, M. R., OHISHI, H., SHIGEMOTO, R. & MUGNAINI, E. (1998) Metabotropic glutamate receptors are associated with non-synaptic appendages of unipolar brush cells in rat cerebellar cortex and cochlear nuclear complex. Journal of Neurocytology 27, 303–327.

    Google Scholar 

  • JAARSMA, D., WENTHOLD, R. J. & MUGNAINI, E. (1995) Glutamate receptor subunits at mossy fiberunipolar brush cell synapses: Light and electron microscopic immunocytochemical study in cerebellar cortex of rat and cat. Journal of Comparative Neurology 357, 145–160.

    Google Scholar 

  • KIND, P. C., KELLY, G. M., FRYER, H. J. L., BLAKEMORE, C. & HOCKFIELD, S. (1997) Phospholipase C-β1 is present in the botrysome, an intermediate compartment-like organelle, and is regulated by visual experience in cat visual cortex. Journal of Neuroscience 17, 1471–1480.

    Google Scholar 

  • LASHER, R. S. & ZAGON, I. S. (1972) The effect of potassium on neuronal differentiation on cultures of dissociated newborn rat cerebellum. Brain Research 41, 428–439.

    Google Scholar 

  • LEVI, G., ALOISI, F., CIOTTI, M. T. & GALLO, V. (1984) Autoradiographic localization and depolarizationinduced release of acid amino acids in differentiating cerebellar granule cell cultures. Brain Research 290, 77–86.

    Google Scholar 

  • LEVI, G., ALOISI, F., CIOTTI, M. T., THANGNIPON, W., KINSBURY, A. & BALAZS, R. (1989) Preparation of 98% pure cerebellar granule cell cultures. In A Dissection and Tissue Culture Manual of the Nervous System (edited by SHAHAR, A., DE VELLIS, J., VERNADAKIS, A. & HABER, B.), pp. 211–214. New York: Alan R. Liss.

    Google Scholar 

  • LEVI, G., WILKIN, G. P., CIOTTI, M. T. & JOHNSTONE, S. (1983) Enrichment of differentiated, stellate astrocytes in cerebellar interneuron cultures as studied by GFAP immunofluorescence and autoradiographic uptake patterns with [3H]D-Aspartate and [3H]GABA. Developmental Brain Research 10, 227–241.

    Google Scholar 

  • MARINI, A. M., STRAUSS, K. I. & JACOBOWITZ, D. M. (1997) Calretinin-containing neurons in rat cerebellar granule cell cultures. Brain Research Bulletin 42, 279–288.

    Google Scholar 

  • MERTZ, K., KOSCHECK, T. & SCHILLING K. (2000) Brain-derived neurotrophic factor modulates dendritic morphology of cerebellar basket and stellate cells: An in vitro study. Neuroscience 97, 303–310.

    Google Scholar 

  • MONTEIRO, R. A. F. (1986) Critical analysis on the nature of synapses en marron of the cerebellar cortex. Journal fûr Hirnforschung 26, 567–576.

    Google Scholar 

  • MORRISON, M. E. & MASON, C. A. (1998) Granule neuron regulation of Purkinje cell development: Striking a balance between neurotrophin and glutamate signaling. Journal of Neuroscience 18, 3563–3573.

    Google Scholar 

  • MUGNAINI, E., DIÑO, M. R. & JAARSMA, D. (1997) The unipolar brush cells of the mammalian cerebellum and cochlear nucleus: Cytology and microcircuitry. Progress in Brain Research 114, 131–150.

    Google Scholar 

  • MUGNAINI, E., FLORIS, A. & WRIGHT-GOSS, M. (1994) Extraordinary synapses of the unipolar brush cell: An electron microscopic study in the rat cerebellum. Synapse 16, 284–311.

    Google Scholar 

  • NUNZI, M. G. & MUGNAINI, E. (1998) Cortex instrinsic mossy fibers in the mammalian cerebellum. Society for Neuroscience Abstracts 24, 668.

    Google Scholar 

  • NUNZI, M. G. & MUGNAINI, E. (1999) UBCs axons form a sizeable portion of the mossy fibers in the vestibulocerebellum. Society for Neuroscience Abstracts 25, 1403.

    Google Scholar 

  • POWELL, S. K., RIVAS, R. J., RODRIGUEZ-BOULAN, E. & HATTEN, M. E. (1997) Development of polarity in cerebellar granule neurons. Journal of Neurobiology 32, 223–236.

    Google Scholar 

  • ROGERS, J. H. (1989) Immunoreactivity for calretinin and other calcium-binding proteins in cerebellum. Neuroscience 31, 711–721.

    Google Scholar 

  • ROSSI, D. J., ALFORD, S., MUGNAINI, E. & SLATER, N. T. (1995) Properties of transmission at a giant glutamatergic synapse in cerebellum: The mossy fiber-unipolar brush cell synapse. Journal of Neurophysiology 74, 24–42.

    Google Scholar 

  • SCHUERGER, R. J., DIÑO, M. R., LIU, N. T., SLATER, N. T. & MUGNAINI, E. (1997) Light and electron microscopic identification of the axon terminals and postsynaptic targets of cerebellar unipolar brush cells. Society for Neuroscience Abstracts 27, 1830.

    Google Scholar 

  • SEKERKOVÁ, G. & MUGNAINI, E. (1997) Prenatal neurogenesis of cerebellar unipolar brush cells studied by bromodeoxyuridine and cell class specific markers. Society for Neuroscience Abstracts 27, 1831.

    Google Scholar 

  • SLATER, N. T., ROSSI, D. J. & KINNEY, G. A. (1997) Physiology of transmission at a giant glutamatergic synapse in cerebellum. Progress in Brain Research 114, 151–163.

    Google Scholar 

  • VOOGD, J., JAARSMA, D. & MARANI, E. (1996) Cerebellum: Chemoarchitecture and anatomy. In Handbook of Chemical Neuroanatomy. Integrated systems of the CNS, part III Cerebellum, Basal Ganglia, Olfactory System (edited by SWANSON, L. W., BJÖRKLUND, A. & HÖKFELT, T.) Vol. 12, pp. 1–369. Amsterdam: Elsevier.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Anelli, R., Dunn, M.E. & Mugnaini, E. Unipolar brush cells develop a set of characteristic features in primary cerebellar cultures. J Neurocytol 29, 129–144 (2000). https://doi.org/10.1023/A:1007108613460

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007108613460

Keywords

Navigation