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. 1981 Jul 1;90(1):1–6. doi: 10.1083/jcb.90.1.1

Immunocytochemical localization of P0 protein in Golgi complex membranes and myelin of developing rat Schwann cells

PMCID: PMC2111837  PMID: 6166623

Abstract

P0 protein, the dominant protein in peripheral nervous system myelin, was studied immunocytochemically in both developing and mature Schwann cells. Trigeminal and sciatic nerves from newborn, 7-d, and adult rats were processed for transmission electron microscopy. Alternating 1- micrometer-thick Epon sections were stained with paraphenylenediamine (PD) or with P0 antiserum according to the peroxidase-antiperoxidase method. To localize P0 in Schwann cell cytoplasm and myelin membranes, the distribution of immunostaining observed in 1-micrometer sections was mapped on electron micrographs of identical areas found in adjacent thin sections. The first P0 staining was observed around axons and/or in cytoplasm of Schwann cells that had established a 1:1 relationship with axons. In newborn nerves, staining of newly formed myelin sheaths was detected more readily with P0 antiserum than with PD. Myelin sheaths with as few as three lamellae could be identified with the light microscope. Very thin sheaths often stained less intensely and part of their circumference frequently was unstained. Schmidt-Lanterman clefts found in more mature sheaths also were unstained. As myelination progressed, intensely stained myelin rings became much more numerous and, in adult nerves, all sheaths were intensely and uniformly stained. Particulate P0 staining also was observed in juxtanuclear areas of Schwann cell cytoplasm. It was most prominent during development, then decreased, but still was detected in adult nerves. The cytoplasmic areas stained by P0 antiserum were rich in Golgi complex membranes.

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Selected References

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  1. Aguayo A. J., Charron L., Bray G. M. Potential of Schwann cells from unmyelinated nerves to produce myelin: a quantitative ultrastructural and radiographic study. J Neurocytol. 1976 Oct;5(8):565–573. doi: 10.1007/BF01175570. [DOI] [PubMed] [Google Scholar]
  2. Baskin D. G., Erlandsen S. L., Parsons J. A. Immunocytochemistry with osmium-fixed tissue. I. Light microscopic localization of growth hormone and prolactin with the unlabeled antibody-enzyme method. J Histochem Cytochem. 1979 Apr;27(4):867–872. doi: 10.1177/27.4.109497. [DOI] [PubMed] [Google Scholar]
  3. Baskin D. G., Erlandsen S. L., Parsons J. A. Influence of hydrogen peroxide or alcoholic sodium hydroxide on the immunocytochemical detection of growth hormone and prolactin after osmium fixation. J Histochem Cytochem. 1979 Sep;27(9):1290–1292. doi: 10.1177/27.9.383831. [DOI] [PubMed] [Google Scholar]
  4. Brockes J. P., Raff M. C., Nishiguchi D. J., Winter J. Studies on cultured rat Schwann cells. III. Assays for peripheral myelin proteins. J Neurocytol. 1980 Feb;9(1):67–77. doi: 10.1007/BF01205227. [DOI] [PubMed] [Google Scholar]
  5. Campagnoni A. T., Carey G. D., Yu Y. T. In vitro synthesis of the myelin basic proteins: subcellular site of synthesis. J Neurochem. 1980 Mar;34(3):677–686. doi: 10.1111/j.1471-4159.1980.tb11197.x. [DOI] [PubMed] [Google Scholar]
  6. Cohen S. R., Guarnieri M. Immunochemical measurement of myelin basic protein in developing rat brain: an index of myelin synthesis. Dev Biol. 1976 Mar;49(1):294–299. doi: 10.1016/0012-1606(76)90276-1. [DOI] [PubMed] [Google Scholar]
  7. Gould R. M. Incorporation of glycoproteins into peripheral nerve myelin. J Cell Biol. 1977 Nov;75(2 Pt 1):326–338. doi: 10.1083/jcb.75.2.326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Greenfield S., Brostoff S., Eylar E. H., Morell P. Protein composition of myelin of the peripheral nervous system. J Neurochem. 1973 Apr;20(4):1207–1216. doi: 10.1111/j.1471-4159.1973.tb00089.x. [DOI] [PubMed] [Google Scholar]
  9. Itoyama Y., Sternberger N. H., Kies M. W., Cohen S. R., Richardson E. P., Jr, Webster H. Immunocytochemical method to identify myelin basic protein in oligodendroglia and myelin sheaths of the human nervous system. Ann Neurol. 1980 Feb;7(2):157–166. doi: 10.1002/ana.410070211. [DOI] [PubMed] [Google Scholar]
  10. Knipe D. M., Baltimore D., Lodish H. F. Separate pathways of maturation of the major structural proteins of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1128–1139. doi: 10.1128/jvi.21.3.1128-1139.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Knipe D. M., Lodish H. F., Baltimore D. Localization of two cellular forms of the vesicular stomatitis viral glycoprotein. J Virol. 1977 Mar;21(3):1121–1127. doi: 10.1128/jvi.21.3.1121-1127.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Peterson R. G., Gruener R. W. Morphological localization of PNS myelin proteins. Brain Res. 1978 Aug 18;152(1):17–29. doi: 10.1016/0006-8993(78)90131-2. [DOI] [PubMed] [Google Scholar]
  13. Peterson R. G., Pease D. C. Myelin embedded in polymerized glutaraldehyde-urea. J Ultrastruct Res. 1972 Oct;41(1):115–132. doi: 10.1016/s0022-5320(72)90042-1. [DOI] [PubMed] [Google Scholar]
  14. Rothman J. E., Bursztyn-Pettegrew H., Fine R. E. Transport of the membrane glycoprotein of vesicular stomatitis virus to the cell surface in two stages by clathrin-coated vesicles. J Cell Biol. 1980 Jul;86(1):162–171. doi: 10.1083/jcb.86.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sternberger L. A., Hardy P. H., Jr, Cuculis J. J., Meyer H. G. The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem. 1970 May;18(5):315–333. doi: 10.1177/18.5.315. [DOI] [PubMed] [Google Scholar]
  16. Sternberger N. H., Itoyama Y., Kies M. W., Webster H deF Immunocytochemical method to identify basic protein in myelin-forming oligodendrocytes of newborn rat C.N.S. J Neurocytol. 1978 Apr;7(2):251–263. doi: 10.1007/BF01217922. [DOI] [PubMed] [Google Scholar]
  17. Sternberger N. H., Itoyama Y., Kies M. W., Webster H. D. Myelin basic protein demonstrated immunocytochemically in oligodendroglia prior to myelin sheath formation. Proc Natl Acad Sci U S A. 1978 May;75(5):2521–2524. doi: 10.1073/pnas.75.5.2521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sternberger N. H., Quarles R. H., Itoyama Y., Webster H. D. Myelin-associated glycoprotein demonstrated immunocytochemically in myelin and myelin-forming cells of developing rat. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1510–1514. doi: 10.1073/pnas.76.3.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Trapp B. D., McIntyre L. J., Quarles R. H., Sternberger N. H., Webster H. D. Immunocytochemical localization of rat peripheral nervous system myelin proteins: P2 protein is not a component of all peripheral nervous system myelin sheaths. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3552–3556. doi: 10.1073/pnas.76.7.3552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Webster H. D., Martin R., O'Connell M. F. The relationships between interphase Schwann cells and axons before myelination: a quantitative electron microscopic study. Dev Biol. 1973 Jun;32(2):401–416. doi: 10.1016/0012-1606(73)90250-9. [DOI] [PubMed] [Google Scholar]
  21. Wiggins R. C., Morell P. Phosphorylation and fucosylation of myelin protein in vitro by sciatic nerve from developing rats. J Neurochem. 1980 Mar;34(3):627–634. doi: 10.1111/j.1471-4159.1980.tb11190.x. [DOI] [PubMed] [Google Scholar]
  22. Wood J. G., Engel E. L. Peripheral nerve glycoproteins and myelin fine structure during development of rat sciatic nerve. J Neurocytol. 1976 Oct;5(8):605–615. doi: 10.1007/BF01175573. [DOI] [PubMed] [Google Scholar]
  23. Wood J. G., McLaughlin B. J. The visualization of concanavalin-A binding sites in the interperiod line of rat sciatic nerve myelin. J Neurochem. 1975 Feb;24(2):233–235. doi: 10.1111/j.1471-4159.1975.tb11870.x. [DOI] [PubMed] [Google Scholar]

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