Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. Feb 1, 1986; 102(2): 384–392.
PMCID: PMC2114066

Emergence of three myelin proteins in oligodendrocytes cultured without neurons

Abstract

Oligodendrocytes, the myelin-forming cells of the central nervous system, were cultured from newborn rat brain and optic nerve to allow us to analyze whether two transmembranous myelin proteins, myelin- associated glycoprotein (MAG) and proteolipid protein (PLP), were expressed together with myelin basic protein (MBP) in defined medium with low serum and in the absence of neurons. Using double label immunofluorescence, we investigated when and where these three myelin proteins appeared in cells expressing galactocerebroside (GC), a specific marker for the oligodendrocyte membrane. We found that a proportion of oligodendrocytes derived from brain and optic nerve invariably express MBP, MAG, and PLP about a week after the emergence of GC, which occurs around birth. In brain-derived oligodendrocytes, MBP and MAG first emerge between the fifth and the seventh day after birth, followed by PLP 1 to 2 d later. All three proteins were confined to the cell body at that time, although an extensive network of GC positive processes had already developed. Each protein shows a specific cytoplasmic localization: diffuse for MBP, mostly perinuclear for MAG, and particulate for PLP. Interestingly, MAG, which may be involved in glial-axon interactions, is the first myelin protein detected in the processes at approximately 10 d after birth. MBP and PLP are only seen in these locations after 15 d. All GC-positive cells express the three myelin proteins by day 19. Simultaneously, numerous membrane and myelin whorls accumulate along the oligodendrocyte surface. The sequential emergence, cytoplasmic location, and peak of expression of these three myelin proteins in vitro follow a pattern similar to that described in vivo and, therefore, are independent of continuous neuronal influences. Such cultures provide a convenient system to study factors regulating expression of myelin proteins.

Full Text

The Full Text of this article is available as a PDF (1.2M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Abney ER, Bartlett PP, Raff MC. Astrocytes, ependymal cells, and oligodendrocytes develop on schedule in dissociated cell cultures of embryonic rat brain. Dev Biol. 1981 Apr 30;83(2):301–310. [PubMed]
  • Abney ER, Williams BP, Raff MC. Tracing the development of oligodendrocytes from precursor cells using monoclonal antibodies, fluorescence-activated cell sorting, and cell culture. Dev Biol. 1983 Nov;100(1):166–171. [PubMed]
  • Agrawal HC, Hartman BK, Shearer WT, Kalmbach S, Margolis FL. Purification and immunohistochemical localization of rat brain myelin proteolipid protein. J Neurochem. 1977 Mar;28(3):495–508. [PubMed]
  • Arnheiter H, Dubois-Dalcq M, Lazzarini RA. Direct visualization of protein transport and processing in the living cell by microinjection of specific antibodies. Cell. 1984 Nov;39(1):99–109. [PubMed]
  • Barbarese E, Pfeiffer SE. Developmental regulation of myelin basic protein in dispersed cultures. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1953–1957. [PMC free article] [PubMed]
  • Barbarese E, Braun PE, Carson JH. Identification of prelarge and presmall basic proteins in mouse myelin and their structural relationship to large and small basic proteins. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3360–3364. [PMC free article] [PubMed]
  • Bologa L, Bisconte JC, Joubert R, Margules S, Herschkowitz N. Proliferative activity and characteristics of immunocytochemically identified oligodendrocytes in embryonic mouse brain cell cultures. Exp Brain Res. 1983;50(1):84–90. [PubMed]
  • Bologa-Sandru L, Siegrist HP, Z'Graggen A, Hofmann K, Wiesmann U, Dahl D, Herschkowitz N. Expression of antigenic markers during the development of oligodendrocytes in mouse brain cell cultures. Brain Res. 1981 Apr 6;210(1-2):217–229. [PubMed]
  • Bradel EJ, Prince FP. Cultured neonatal rat oligodendrocytes elaborate myelin membrane in the absence of neurons. J Neurosci Res. 1983;9(4):381–392. [PubMed]
  • Brockes JP, Fryxell KJ, Lemke GE. Studies on cultured Schwann cells: the induction of myelin synthesis, and the control of their proliferation by a new growth factor. J Exp Biol. 1981 Dec;95:215–230. [PubMed]
  • Campagnoni AT, Hunkeler MJ. Synthesis of the myelin proteolipid protein in the developing mouse brain. J Neurobiol. 1980 Jul;11(4):355–364. [PubMed]
  • Campagnoni AT, Carey GD, Yu YT. In vitro synthesis of the myelin basic proteins: subcellular site of synthesis. J Neurochem. 1980 Mar;34(3):677–686. [PubMed]
  • Carey DJ, Eldridge CF, Cornbrooks CJ, Timpl R, Bunge RP. Biosynthesis of type IV collagen by cultured rat Schwann cells. J Cell Biol. 1983 Aug;97(2):473–479. [PMC free article] [PubMed]
  • Colman DR, Kreibich G, Frey AB, Sabatini DD. Synthesis and incorporation of myelin polypeptides into CNS myelin. J Cell Biol. 1982 Nov;95(2 Pt 1):598–608. [PMC free article] [PubMed]
  • Eccleston PA, Silberberg DH. The differentiation of oligodendrocytes in a serum-free hormone-supplemented medium. Brain Res. 1984 Sep;318(1):1–9. [PubMed]
  • Fritz RB, Chou CH. Epitopes of peptide 43-88 of guinea pig myelin basic protein: localization with monoclonal antibodies. J Immunol. 1983 May;130(5):2180–2182. [PubMed]
  • Fry JM, Lisak RP, Manning MC, Silberberg DH. Serological techniques for detection of antibody to galactocerebroside. J Immunol Methods. 1976;11(2):185–193. [PubMed]
  • Guentert-Lauber B, Monnet-Tschudi F, Omlin FX, Favrod P, Honegger P. Serum-free aggregate cultures of rat CNS glial cells: biochemical, immunocytochemical and morphological characterization. Dev Neurosci. 1985;7(1):33–44. [PubMed]
  • Hartman BK, Agrawal HC, Agrawal D, Kalmbach S. Development and maturation of central nervous system myelin: comparison of immunohistochemical localization of proteolipid protein and basic protein in myelin and oligodendrocytes. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4217–4220. [PMC free article] [PubMed]
  • Hartman BK, Agrawal HC, Kalmbach S, Shearer WT. A comparative study of the immunohistochemical localization of basic protein to myelin and oligodendrocytes in rat and chicken brain. J Comp Neurol. 1979 Nov 15;188(2):273–290. [PubMed]
  • Ilyas AA, Quarles RH, MacIntosh TD, Dobersen MJ, Trapp BD, Dalakas MC, Brady RO. IgM in a human neuropathy related to paraproteinemia binds to a carbohydrate determinant in the myelin-associated glycoprotein and to a ganglioside. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1225–1229. [PMC free article] [PubMed]
  • Laursen RA, Samiullah M, Lees MB. The structure of bovine brain myelin proteolipid and its organization in myelin. Proc Natl Acad Sci U S A. 1984 May;81(9):2912–2916. [PMC free article] [PubMed]
  • McCarthy KD, de Vellis J. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol. 1980 Jun;85(3):890–902. [PMC free article] [PubMed]
  • McMorris FA. Cyclic AMP induction of the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphohydrolase in rat oligodendrocytes. J Neurochem. 1983 Aug;41(2):506–515. [PubMed]
  • Mirsky R, Winter J, Abney ER, Pruss RM, Gavrilovic J, Raff MC. Myelin-specific proteins and glycolipids in rat Schwann cells and oligodendrocytes in culture. J Cell Biol. 1980 Mar;84(3):483–494. [PMC free article] [PubMed]
  • Mithen FA, Wood PM, Agrawal HC, Bunge RP. Immunohistochemical study of myelin sheaths formed by oligodendrocytes interacting with dissociated dorsal root ganglion neurons in culture. Brain Res. 1983 Feb 28;262(1):63–69. [PubMed]
  • Norton WT, Farooq M, Fields KL, Raine CS. The long term culture of bulk-isolated bovine oligodendroglia from adult brain. Brain Res. 1983 Jul 4;270(2):295–310. [PubMed]
  • Omlin FX, Webster HD, Palkovits CG, Cohen SR. Immunocytochemical localization of basic protein in major dense line regions of central and peripheral myelin. J Cell Biol. 1982 Oct;95(1):242–248. [PMC free article] [PubMed]
  • Pfeiffer SE, Barbarese E, Bhat S. Noncoordinate regulation of myelinogenic parameters in primary cultures of dissociated fetal rat brain. J Neurosci Res. 1981;6(3):369–380. [PubMed]
  • Sobue G, Pleasure D. Schwann cell galactocerebroside induced by derivatives of adenosine 3',5'-monophosphate. Science. 1984 Apr 6;224(4644):72–74. [PubMed]
  • Poduslo SE, Miller K, Wolinsky JS. The production of a membrane by purified oligodendroglia maintained in culture. Exp Cell Res. 1982 Jan;137(1):203–215. [PubMed]
  • Poduslo JF, Berg CT, Dyck PJ. Schwann cell expression of a major myelin glycoprotein in the absence of myelin assembly. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1864–1866. [PMC free article] [PubMed]
  • Poduslo SE, Miller K, Zoller S. Purification and maintenance in culture of oligodendroglia from human multiple sclerosis brain. J Neurol Sci. 1982 Jun;54(3):395–400. [PubMed]
  • Quarles RH. Glycoproteins from central and peripheral myelin. Prog Clin Biol Res. 1980;49:55–77. [PubMed]
  • Quarles RH. Myelin-associated glycoprotein in development and disease. Dev Neurosci. 1983;6(6):285–303. [PubMed]
  • Raff MC, Fields KL, Hakomori SI, Mirsky R, Pruss RM, Winter J. Cell-type-specific markers for distinguishing and studying neurons and the major classes of glial cells in culture. Brain Res. 1979 Oct 5;174(2):283–308. [PubMed]
  • Raff MC, Miller RH, 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. [PubMed]
  • Raff MC, Mirsky R, Fields KL, Lisak RP, Dorfman SH, Silberberg DH, Gregson NA, Leibowitz S, Kennedy MC. Galactocerebroside is a specific cell-surface antigenic marker for oligodendrocytes in culture. Nature. 1978 Aug 24;274(5673):813–816. [PubMed]
  • Raff MC, Williams BP, Miller RH. The in vitro differentiation of a bipotential glial progenitor cell. EMBO J. 1984 Aug;3(8):1857–1864. [PMC free article] [PubMed]
  • Ranscht B, Clapshaw PA, Price J, Noble M, Seifert W. Development of oligodendrocytes and Schwann cells studied with a monoclonal antibody against galactocerebroside. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2709–2713. [PMC free article] [PubMed]
  • Richardson CD, Berkovich A, Rozenblatt S, Bellini WJ. Use of antibodies directed against synthetic peptides for identifying cDNA clones, establishing reading frames, and deducing the gene order of measles virus. J Virol. 1985 Apr;54(1):186–193. [PMC free article] [PubMed]
  • Roussel G, Nussbaum JL. Comparative localization of Wolfgram W1 and myelin basic proteins in the rat brain during ontogenesis. Histochem J. 1981 Nov;13(6):1029–1047. [PubMed]
  • Roussel G, Sensenbrenner M, Labourdette G, Wittendorp-Rechenmann E, Pettmann B, Nussbaum JL. An immunohistochemical study of two myelin-specific proteins in enriched oligodendroglial cell cultures combined with an autoradiographic investigation using [3H]thymidine. Brain Res. 1983 Jun;284(2-3):193–204. [PubMed]
  • Sarliève LL, Fabre M, Susz J, Matthieu JM. Investigations on myelination in vitro: IV. "Myelin-like" or premyelin structures in cultures of dissociated brain cells from 14--15-day-old embryonic mice. J Neurosci Res. 1983;10(2):191–210. [PubMed]
  • Sternberger NH, Itoyama Y, Kies MW, Webster HD. 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. [PMC free article] [PubMed]
  • Sternberger NH, Itoyama Y, Kies MW, 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. [PubMed]
  • Sternberger NH, Quarles RH, Itoyama Y, Webster HD. 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. [PMC free article] [PubMed]
  • Stoffel W, Hillen H, Giersiefen H. Structure and molecular arrangement of proteolipid protein of central nervous system myelin. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5012–5016. [PMC free article] [PubMed]
  • Temple S, Raff MC. Differentiation of a bipotential glial progenitor cell in a single cell microculture. Nature. 1985 Jan 17;313(5999):223–225. [PubMed]
  • Tennekoon GI, Cohen SR, Price DL, McKhann GM. Myelinogenesis in optic nerve. A morphological, autoradiographic, and biochemical analysis. J Cell Biol. 1977 Mar;72(3):604–616. [PMC free article] [PubMed]
  • Townsend LE, Agrawal D, Benjamins JA, Agrawal HC. In vitro acylation of rat brain myelin proteolipid protein. J Biol Chem. 1982 Aug 25;257(16):9745–9750. [PubMed]
  • Trapp BD, Quarles RH. Presence of the myelin-associated glycoprotein correlates with alterations in the periodicity of peripheral myelin. J Cell Biol. 1982 Mar;92(3):877–882. [PMC free article] [PubMed]
  • Trapp BD, Quarles RH. Immunocytochemical localization of the myelin-associated glycoprotein. Fact or artifact? J Neuroimmunol. 1984 Jul;6(4):231–249. [PubMed]
  • Trapp BD, Webster HD, Johnson D, Quarles RH, Cohen SR, Murray MR. Myelin formation in rotation-mediated aggregating cell cultures: immunocytochemical, electron microscopic, and biochemical observations. J Neurosci. 1982 Jul;2(7):986–993. [PubMed]
  • Trapp BD, Quarles RH, Suzuki K. Immunocytochemical studies of quaking mice support a role for the myelin-associated glycoprotein in forming and maintaining the periaxonal space and periaxonal cytoplasmic collar of myelinating Schwann cells. J Cell Biol. 1984 Aug;99(2):594–606. [PMC free article] [PubMed]
  • Wood PM, Williams AK. Oligodendrocyte proliferation and CNS myelination in cultures containing dissociated embryonic neuroglia and dorsal root ganglion neurons. Brain Res. 1984 Feb;314(2):225–241. [PubMed]
  • Wood P, Okada E, Bunge R. The use of networks of dissociated rat dorsal root ganglion neurons to induce myelination by oligodencrocytes in culture. Brain Res. 1980 Aug 25;196(1):247–252. [PubMed]
  • Zeller NK, Behar TN, Dubois-Dalcq ME, Lazzarini RA. The timely expression of myelin basic protein gene in cultured rat brain oligodendrocytes is independent of continuous neuronal influences. J Neurosci. 1985 Nov;5(11):2955–2962. [PubMed]
  • Zurbriggen A, Vandevelde M, Steck A, Angst B. Myelin-associated glycoprotein is produced before myelin basic protein in cultured oligodendrocytes. J Neuroimmunol. 1984 Feb;6(1):41–49. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...