Abstract
To identify new proteins, which are expressed in oligodendrocytes and which may have a functional role in myelination, a rat oligodendrocyte cDNA library was screened using differential and subtractive screening techniques. Ten clones that have elevated levels of expression in brain were isolated. Two of these clones were characterized further and one clone, pC26.H2, was found to be closely related to mouse stearoyl-CoA desaturase 2 (SCD2), which catalyzes the synthesis of unsaturated fatty acid. From Northern blot and in situ hybridization studies, SCD2 mRNA was expressed primarily in brain with lower levels found in lung and spleen. In brain sections, SCD2 mRNA was found primarily in oligodendrocytes, although mRNA was detected at a low level in neurons, in particular in Purkinje cells in the cerebellum. Northern blot analysis of the other clone, p973.HB, indicated that it was expressed more selectively in brain. In mixed glial cultures oligodendrocytes were the only cells that expressed this mRNA, whereas in brain, neurons expressed this mRNA at a higher level than in oligodendrocytes. This clone is being actively pursued because of its unique expression exclusively in oligodendrocytes and neurons.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Huxley, A. F., and Staempfli, R. 1949. Evidence for saltatory conduction in peripheral myelinated nerve fibres. J. Physiol. 108:315–339.
Raine, C. S. 1984. Morphology of myelin and myelination, Pages 1–50,in P. Morell (ed.), Myelin, Plenum Press, New York.
Mikoshiba, K., Okano, H., Tamura, T., and Ikenaka, K. 1991. Structure and function of myelin protein genes. Annu. Rev. Neurosci. 14:201–217.
Cambi, F., Lees, M. B., Williams, R. M., and Macklin, W. B. 1983. Chronic experimental allergic encephalomyelitis produced by bovine proteolipid apoprotein: Immunological studies in rabbits. Ann. Neurol. 13:303–308.
Tuohy, V. K., Lu, Z., Sobel, R. A., Laursen, R. A., and Lees, M. B. 1988. A synthetic peptide from myelin proteolipid protein induces experimental aliergic encephalomyelitis. J. Immun. 141:1126–1130.
Watson, J. B., and Margulies, J. E. 1993. Differential cDNA screening strategies to identify novel stage-specific proteins in the developing mammalian brain. Dev. Neurosci. In press.
Watson, J. B., Battenberg, E. F., Wong, K. K., Bloom, F. E., and Sutcliffe, J. G. 1990. Subtractive cDNA cloning of RC3, a rodent cortex-enriched mRNA encoding a novel 78 residue protein. J. Neurosci. Res. 26:397–408.
Travis, G. H., Brennan, M. B., Danielson, P. E., Kozak, C. A., and Sutcliffe, J. G. 1989. Identification of a photoreceptor-specific mRNA encoded by the gene responsible for retinal degeneration slow (rds). Nature. 338:70–73.
Welcher, A. A., Suter, U., De Leon, M., Snipes, G. J., and Shooter, E. M. 1991. A myelin protein is encoded by the homologue of a growth arrest-specific gene. Proc. Natl. Acad. Sci. USA 88:7195–7199.
Suter, U., Welcher, A. A., Özcelik, T., Snipes, G. J., Kosaras, B., Francke, U., Billings-Gagliardi, S., Sidman, R. L., and Shooter, E. M. 1992. Trembler mouse carries a point mutation in a myelin gene. Nature. 356:241–244.
Patel, P. I., Roa, B. B., Welcher, A. A., Schoener-Scott, R., Trask, B. J., Pentao, L., Snipes, G. J., Garcia, C. A., Francke, U., Schooter, E. M. et al. 1992. The gene for the peripheral myelin protein PMP-22 is a candidate for Charcot-Marie-Tooth disease type 1A. Nat. Genet. 1:159–165.
McCarthy, K. D., and de Vellis, J. 1980. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J. Cell Biol. 85:890–902.
Watson, J. B., Coulter II, P. M., and Sutcliffe, J. G. 1992. Identification of genes enriched in expression in mammalian neostriatum and neocortex. Brain Dysfunct. 5:94–105.
Dworkin, M. B., and Dawid, I. B. 1980. Use of a cloned library for the study of abundant poly (A)+-RNA duringxenopus laevis development. Develop. Biol. 76:449–464.
Roach, A., Boylan, K., Horvath, S., Prusiner, S. B., and Hood, L. E. 1983. Characterization of cloned cDNA representing rat myelin basic protein: absence of expression in brain of shiverer mutant mice. Cell 34:799–806.
LeVine, S. M., Wong, D., and Macklin, W. B. 1990. Developmental expression of proteolipid protein and DM20 mRNAs and proteins in the rat brain. Dev. Neurosci. 12:235–250.
Bernier, L., Alvarez, F., Norgard, E. M., Raible, D. W., Mentaberry, A., Schembri, J. G., Sabatini, D. D., and Colman, D. R. 1987. Molecular cloning of a 2′,3′-cyclic nucleotide 3′-phosphodiesterase: mRNAs with different 5′ ends encode the same set of proteins in nervous and lymphoid tissue. J. Neurosci. 7:2703–2710.
Salzer, J. L., Holmes, W. P., and Colman, D. R. 1987. The amino acid sequence of the myelin-associated glycoproteins: homology to the immunoglobulin gene superfamily. J. Cell Biol. 104:957–965.
Travis, G. H. and Sutcliffe, J. G. 1988. Phenol emulsion-enhanced DNA-driven subtractive cDNA cloning: isolation of low-abundance monkey cortex-specific mRNAs. Proc. Natl. Acad. Sci. USA. 85:1696–1700.
Travis, G. H., Milner, R. J., and Sutcliffe, J. G. 1990. Preparation and use of subtractive cDNA hybridization probes for cDNA cloning, Pages 49–78,in Boulton, A. A., Baker, G. B., and Campagnoni, A. T. (eds.) Neuromethods: Molecular Neurobiological Techniques. vol. 16, Clifton, The Humana Press.
Wight, P. A., Duchala, C. S., Readhead, C., and Macklin, W. B. 1993. A myelin proteolipid protein-LacZ fusion protein is developmentally regulated and targeted to the myelin membrane in transgenic mice. J. Cell Biol. 123:443–454.
Asotra, K., and Macklin, W. B. 1993. Protein kinase C activity modulates myelin gene expression in enriched oligodendrocytes. J. Neurosci. Res. 34:571–588.
Amur-Umarjee, S., Phan, T. and Campagnoni, A. T. 1993. Myelin basic protein mRNA translocation in oligodendrocytes is inhibited by astrocytes in vitro. J. Neurosci. Res. 36:99–110.
Bartsch, S., Bartsch, U., Dörries, U., Faissner, A., Weller, A., Ekblom, P., and Schachner, M. 1992. Expression of tenascin in the developing and adult cerebellar cortex. J. Neurosci. 12:736–749.
Fuss, S., Wintergerst, E. S., Bartsch, U., and Schachner, M. 1993. Molecular characterization and in situ mRNA localization of the neural recognition molecule J1-160/180: a modular structure similar to tenascin. J. Cell Biol. 120:1237–1249.
Sanger, F., Nicklen, S., and Coulson, A. R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA. 74:5463–5467.
Ranscht, B., Clapshaw, P. A., Price, J., Noble, M., Seifert, W. 1982. Development of oligodendrocytes and Schwann cells studied with a monoclonal antibody against galactocerebroside. Proc. Natl. Acad. Sci. USA. 79:2709–2713.
Forss-Petter, S., Danielson, P., and Sutcliffe, J. G. 1986. Neuronspecific enolase: complete structure of rat mRNA, multiple transcriptional start sites, and evidence suggesting post-transcriptional control. J. Neurosci. Res. 16:141–156.
Miller, F. D., Naus, C. C. G., Higgins, G. A., Bloom, F. E., and Milner, R. J. 1987. Developmentally regulated rat brain mRNAs: Molecular and anatomical characterization. J. Neurosci. 7:2433–2444.
Fischer, I., Konola, J., and Cochary, E. 1990. Microtubule associate protein (MAP1B) is present in cultured oligodendrocytes and co-localizes with tubulin. J. Neurosci. Res. 27:112–124.
Schoenfeld, T. A., McKerracher, L., Obar, R., and Vallee, R. B. 1989. MAP1A and MAP1B are structurally related microtubule associated proteins with distinct developmental patterns in the CNS. J. Neurosci. 9:1712–1730.
Enoch, H. G., Catalá, A., and Strittmatter, P. 1976. Mechanism of rat liver microsomal stearyl-CoA desaturase. Studies of the substrate specificity, enzyme-substrate interactions, and the function of lipid. J. Biol. Chem. 251:5095–5103.
Thiede, M. A., Ozols, J., and Strittmatter, P. 1986. Construction and sequence of cDNA for rat liver stearyl coenzyme A desaturase. J. Biol. Chem. 261:13230–13235.
Ntambi, J. M., Buhrow, S. A., Kaestner, K. H., Christy, R. J., Sibley, E., Kelly, Jr., T. J., and Lane, M. D. 1988. Differentiation-induced gene expression in 3T3-L1 preadipocytes. J. Biol Chem. 263:17291–17300.
Kaestner, K. H., Ntambi, J. M., Kelly, Jr., T. J., and Lane, M. D. 1989. Differentiation-induced gene expression in 3T3-L1 preadipocytes. J. Biol. Chem. 264:14755–14761.
Mihara, K. 1990. Structure and regulation of rat liver microsomal stearoyl-CoA desaturase gene. J. Biochem. 108:1022–1029.
Galli, C., Trzeciak, H. I., and Paoletti, R. 1972. Effects of essential fatty acid deficiency on myelin and various subcellular structures in rat brain. J. Neurochem. 19:1863–1867.
Tebbey, P. W., and Buttke, T. 1992. Stearoyl-CoA desaturase gene expression in lymphocytes. Biochem. & Biophys. Res. Comm. 186:531–536.
DeWille, J. W., and Farmer, S. J. 1992. Postnatal dietary fat influences mRNAs involved in myelination. Dev. Neurosci. 14:61–68.
Nave, K.-A., and Milner, R. J. 1989. Proteolipid proteins: structure and genetic expression in normal and myelin-deficient mutant mice. Crit. Rev. Neurobiol. 5:65–91.
Baumann, N. 1980. Neurological mutations affecting myelination. Inserm Symposium 14, Elsevier, Amsterdam, 565 pp.
DeWille, J. W., and Farmer, S. J. 1992. Quaking phenotype influences brain lipid-related mRNA levels. Neurosci. Lett. 141:195–198.
Sórg, B. A., Smith, M. M., and Campagnoni, A. T. 1987. Developmental expression of the myelin proteolipid protein and basic protein mRNAs in normal and dysmyelinating mutant mice. J. Neurochem. 49:1146–1154.
Author information
Authors and Affiliations
Additional information
Special issue dedicated to Dr. Marjorie B. Lees.
Rights and permissions
About this article
Cite this article
Baba, H., Fuss, B., Watson, J.B. et al. Identification of novel mRNAs expressed in oligodendrocytes. Neurochem Res 19, 1091–1099 (1994). https://doi.org/10.1007/BF00968721
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00968721