Abstract
The adult rat cerebellum has minimal enkephalin immunoreactivity and is devoid of opiate-binding activity. Using novel monoclonal antibodies to the mammalian enkephalin precursor, we describe the immunofluorescent detection of proenkephalin, in the absence of mature enkephalin peptides, in subpopulations of rat cerebellar neurons and astrocytes. In cryostat sections, neurons that express proenkephalin include Golgi cells, macroneurons within deep cerebellar nuclei and a subpopulation of Purkinje cells. Proenkephalin messenger RNA and protein are present in subpopulations of both grey and white matter astrocytes, but not Bergmann glia. In dissociated glial culture, proenkephalin is expressed in process-bearing astrocytes, apparently in association with a subset of intermediate filaments. Proenkephalin within astrocytes is not seen until the second postnatal week and increases through to adulthood. Neuropeptide gene expression adds to the growing range of neuronal-type properties glial cells can display.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abd-el-Basset E. M., Kalnins V. I., Ahmed I., Fedoroff S. A 48 kilodalton intermediate filament associated protein (IFAP) in reactive-like astrocytes induced by dibutyryl cyclic AMP in culture and in reactive astrocytes in situ. J Neuropathol Exp Neurol. 1989 May;48(3):245–254. doi: 10.1097/00005072-198905000-00002. [DOI] [PubMed] [Google Scholar]
- Barres B. A., Chun L. L., Corey D. P. Ion channel expression by white matter glia: I. Type 2 astrocytes and oligodendrocytes. Glia. 1988;1(1):10–30. doi: 10.1002/glia.440010104. [DOI] [PubMed] [Google Scholar]
- Birch N. P., Christie D. L. Characterization of the molecular forms of proenkephalin in bovine adrenal medulla and rat adrenal, brain, and spinal cord with a site-directed antiserum. J Biol Chem. 1986 Sep 15;261(26):12213–12221. [PubMed] [Google Scholar]
- Comb M., Seeburg P. H., Adelman J., Eiden L., Herbert E. Primary structure of the human Met- and Leu-enkephalin precursor and its mRNA. Nature. 1982 Feb 25;295(5851):663–666. doi: 10.1038/295663a0. [DOI] [PubMed] [Google Scholar]
- Eiden L. E. The enkephalin-containing cell: strategies for polypeptide synthesis and secretion throughout the neuroendocrine system. Cell Mol Neurobiol. 1987 Dec;7(4):339–352. doi: 10.1007/BF00733787. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ibuki T., Okamura H., Miyazaki M., Kimura H., Yanaihara N., Ibata Y. Colocalization of GABA and [Met]enkephalin-Arg6-Gly7-Leu8 in the rat cerebellum. Neurosci Lett. 1988 Aug 31;91(2):131–135. doi: 10.1016/0304-3940(88)90756-2. [DOI] [PubMed] [Google Scholar]
- Johnstone S. R., Levi G., Wilkin G. P., Schneider A., Ciotti M. T. Subpopulations of rat cerebellar astrocytes in primary culture: morphology, cell surface antigens and [3H]GABA transport. Brain Res. 1986 Jan;389(1-2):63–75. doi: 10.1016/0165-3806(86)90173-2. [DOI] [PubMed] [Google Scholar]
- Legon S., Glover D. M., Hughes J., Lowry P. J., Rigby P. W., Watson C. J. The structure and expression of the preproenkephalin gene. Nucleic Acids Res. 1982 Dec 20;10(24):7905–7918. doi: 10.1093/nar/10.24.7905. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levi G., Gallo V., Ciotti M. T. Bipotential precursors of putative fibrous astrocytes and oligodendrocytes in rat cerebellar cultures express distinct surface features and "neuron-like" gamma-aminobutyric acid transport. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1504–1508. doi: 10.1073/pnas.83.5.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melner M. H., Low K. G., Allen R. G., Nielsen C. P., Young S. L., Saneto R. P. The regulation of proenkephalin expression in a distinct population of glial cells. EMBO J. 1990 Mar;9(3):791–796. doi: 10.1002/j.1460-2075.1990.tb08175.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noda M., Furutani Y., Takahashi H., Toyosato M., Hirose T., Inayama S., Nakanishi S., Numa S. Cloning and sequence analysis of cDNA for bovine adrenal preproenkephalin. Nature. 1982 Jan 21;295(5846):202–206. doi: 10.1038/295202a0. [DOI] [PubMed] [Google Scholar]
- Pittius C. W., Kley N., Loeffler J. P., Höllt V. Quantitation of proenkephalin A messenger RNA in bovine brain, pituitary and adrenal medulla: correlation between mRNA and peptide levels. EMBO J. 1985 May;4(5):1257–1260. doi: 10.1002/j.1460-2075.1985.tb03769.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raff M. C., Abney E. R., Cohen J., Lindsay R., Noble M. Two types of astrocytes in cultures of developing rat white matter: differences in morphology, surface gangliosides, and growth characteristics. J Neurosci. 1983 Jun;3(6):1289–1300. doi: 10.1523/JNEUROSCI.03-06-01289.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raff M. C., Miller R. H., Noble M. A glial progenitor cell that develops in vitro into an astrocyte or an oligodendrocyte depending on culture medium. Nature. 1983 Jun 2;303(5916):390–396. doi: 10.1038/303390a0. [DOI] [PubMed] [Google Scholar]
- Shinoda H., Marini A. M., Cosi C., Schwartz J. P. Brain region and gene specificity of neuropeptide gene expression in cultured astrocytes. Science. 1989 Jul 28;245(4916):415–417. doi: 10.1126/science.2569236. [DOI] [PubMed] [Google Scholar]
- Shivers B. D., Harlan R. E., Romano G. J., Howells R. D., Pfaff D. W. Cellular localization of proenkephalin mRNA in rat brain: gene expression in the caudate-putamen and cerebellar cortex. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6221–6225. doi: 10.1073/pnas.83.16.6221. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spruce B. A., Jackson S., Lowry P. J., Lane D. P., Glover D. M. Monoclonal antibodies to a proenkephalin A fusion peptide synthesized in Escherichia coli recognize novel proenkephalin A precursor forms. J Biol Chem. 1988 Dec 25;263(36):19788–19795. [PubMed] [Google Scholar]
- Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
- Tsang D., Ng S. C., Ho K. P., Ho W. K. Ontogenesis of opiate binding sites and radioimmunoassayable beta-endorphin and enkephalin in regions of rat brain. Brain Res. 1982 Nov;281(3):257–261. doi: 10.1016/0165-3806(82)90124-9. [DOI] [PubMed] [Google Scholar]
- Usowicz M. M., Gallo V., Cull-Candy S. G. Multiple conductance channels in type-2 cerebellar astrocytes activated by excitatory amino acids. Nature. 1989 Jun 1;339(6223):380–383. doi: 10.1038/339380a0. [DOI] [PubMed] [Google Scholar]
- Vilijn M. H., Vaysse P. J., Zukin R. S., Kessler J. A. Expression of preproenkephalin mRNA by cultured astrocytes and neurons. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6551–6555. doi: 10.1073/pnas.85.17.6551. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilkin G. P., Levi G., Johnstone S. R., Riddle P. N. Cerebellar astroglial cells in primary culture: expression of different morphological appearances and different ability to take up [3H]D-aspartate and [3H]GABA. Brain Res. 1983 Nov;312(2):265–277. doi: 10.1016/0165-3806(83)90143-8. [DOI] [PubMed] [Google Scholar]
- Williams B. P., Abney E. R., Raff M. C. Macroglial cell development in embryonic rat brain: studies using monoclonal antibodies, fluorescence activated cell sorting, and cell culture. Dev Biol. 1985 Nov;112(1):126–134. doi: 10.1016/0012-1606(85)90126-5. [DOI] [PubMed] [Google Scholar]
- Yoshikawa K., Sabol S. L. Glucocorticoids and cyclic AMP synergistically regulate the abundance of preproenkephalin messenger RNA in neuroblastoma-glioma hybrid cells. Biochem Biophys Res Commun. 1986 Aug 29;139(1):1–10. doi: 10.1016/s0006-291x(86)80071-7. [DOI] [PubMed] [Google Scholar]
- Yoshikawa K., Williams C., Sabol S. L. Rat brain preproenkephalin mRNA. cDNA cloning, primary structure, and distribution in the central nervous system. J Biol Chem. 1984 Nov 25;259(22):14301–14308. [PubMed] [Google Scholar]
- Zagon I. S., Rhodes R. E., McLaughlin P. J. Distribution of enkephalin immunoreactivity in germinative cells of developing rat cerebellum. Science. 1985 Mar 1;227(4690):1049–1051. doi: 10.1126/science.3883485. [DOI] [PubMed] [Google Scholar]