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Cell shape and motility of oligodendrocytes cultured without neurons

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Summary

Oligodendrocytes, the myelin-forming cells of the central nervous system (CNS), were cultured from newborn rat brain and optic nerve to study how they differentiate in vitro in the absence of neurons. By use of galactocerebroside (GC) as a reference marker, the development of the cell phenotype was studied with video-enhanced differential interference contrast microscopy, immunofluorescence and electron microscopy. After a few days in culture, oligodendrocytes extend 5 to 10 main processes that are very rich in microtubules, but they did not stain with a monoclonal antibody reacting with all known classes of intermediate filaments. The number of processes can vary with the substrate on which the cells are grown; fewer processes form on laminin than on polylysine coated glass. Oligodendrocytes, in a fashion similar to that of neurons appear to keep their body immobile while the long processes grow. However, while neurons display motile activities mostly at the end of the cell processes called growth cones, the oligodendrocytes display motile, actin rich filopodia and lamellipodia along the entire length of all processes. The outgrowth of motile processes from oligodendrocytes sometimes occurs preferentially towards neighboring astrocytes. Oligodendrocyte processes display intense bidirectional movement of cytoplasmic organelles. Movement of surface components also occurs since GC molecules cross-linked by antibodies move from the processes towards the cell body. Thus, oligodendrocytes cultured without neurons develop on schedule a complex phenotype similar to their in vivo counterpart. In addition, their processes are capable of specific motile activities which may function in vivo to find the target axon and to transport myelin membrane components at the site of myelin assembly.

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Abbreviations

(CNS):

Central nervous system

(DIC):

Differential interference contrast

(GC):

Galactocerebroside

(GFA) protein:

Glial fibrillary acidic

(NSE):

Neuron-specific enolase

References

  • Abercrombie M, Heaysman JEM, Pegrum SM (1970) The locomotion of fibroblasts in culture. I. Movements of the leading edge. Exp Cell Res 59:393–398

    Google Scholar 

  • Albrecht-Buehler G (1976a) Filopodia in spreading 3T3 cells. Do they have a substrate-exploring function? J Cell Biol 69:275–286

    Google Scholar 

  • Albrecht-Buehler G (1976b) The function of filopodia in spreading 3T3 mouse fibroblasts. In: Goldman R, Pollard T, Rosenbaum J (eds) Cell motility. Cold Spring Harbor Laboratory, New York, pp 247–264

    Google Scholar 

  • Allen RD, Allen NS (1982) Video-enhanced microscopy with a computer frame memory. J Micros 129:3–17

    Google Scholar 

  • Allen RD, Allen NS, Travis JL (1981) Video-enhanced contrast, differential interference contrast (AVEC-DIC) microscopy: a new method capable of analyzing microtubule related motility in the reticulopodial network of Allogromia laticollaris. Cell Motil 1:291–302

    Google Scholar 

  • Bray D (1982) Filopodial contraction and growth cone guidance. In: Bellairs R (ed) Cell behavior. Cambridge University Press, Cambridge England, pp 299

    Google Scholar 

  • Bretscher MS (1984) Endocytosis: relation to capping and cell locomotion. Science 224:681–686

    Google Scholar 

  • DeRosier D, Edds KT (1980) Evidence for fascin crosslinks between actin filaments in coelomocyte filopodia. Exp Cell Res 95:425–439

    Google Scholar 

  • Dubois-Dalcq M, Behar T, Hudson L, Lazzarini RA (1986) Timely emergence of three myelin proteins in oligodendrocytes. J Cell Biol (In press)

  • Eccleston PA, Silberberg DH (1984) The differentiation of oligodendrocytes in a serum-free, hormone supplemented medium. Dev Brain Res 16:1–9

    Google Scholar 

  • Edds KT, Chambers C, Allen RD (1983) Coelomocyte motility. Cell Motil 3:113–121

    Google Scholar 

  • Euteneuer U, Schliwa M (1984) Persistent, directional motility of cells and cytoplasmic fragments in the absence of microtubules. Nature 310:58–61

    Google Scholar 

  • Gonatas NK, Hirayama M, Steiber A, Silberberg DH (1982) The ultrastructure of isolated rat oligodendroglial cell cultures. J Neurocytol 11:997–1008

    Google Scholar 

  • Hayden JH, Allen RD (1984) Detection of single microtubules in living cells: particle transport can occur in both directions along the same microtubules. J Cell Biol 99:1785–1793

    Google Scholar 

  • Herman IM, Crisona NJ, Pollard TD (1981) Relation between cell activity and the distribution of cytoplasmic actin and myosin. J Cell Biol 90:84–91

    Google Scholar 

  • Inoue S (1981) Video image processing greatly enhances contrast, quality and speed in polarization-based microscopy. J Cell Biol 89:364–356

    Google Scholar 

  • Kachar B (1985) Direct visualization of organelle movement along actin cables dissociated from characean algae. Science 227:1355–1357

    Google Scholar 

  • Kachar B, Bridgman PC, Reese TS (1984) Structural relationship of moving cytoplasmic organelles to microtubules in the foraminifer, Allogromia. J Cell Biol 99:50a

    Google Scholar 

  • Kilmartin JV, Wright B, Milstein C (1982) Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol 93:576–582

    Google Scholar 

  • Klymkovsky MW, Miller RH, Lane EB (1983) Morphology behaviour and interaction of cultured epithelial cells after the antibody induced disruption of keratin filament organization. J Cell Biol 96:494–509

    Google Scholar 

  • Knobler RL, Dubois-Dalcq M, Haspel MV, Claysmith AP, Lampert PW, Oldstone MBA (1981) Selective localization of wild type and mutant mouse hepatitis virus (JHM strain) antigens in CNS tissue by fluorescence, light and electron microscopy. J Neuroimmunol 1:81–92

    Google Scholar 

  • Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 282:249–256

    Google Scholar 

  • Liesi P, Dahl D, Vaheri A (1983) Laminin is produced by early rat astrocytes in primary culture. J Cell Biol 96:920–924

    Google Scholar 

  • Lumsden CE, Pomerat CM (1951) Normal oligodendrocytes in tissue culture. Exp Cell Res 2:103–114

    Google Scholar 

  • Luther PW, Peng HB, Lin JJ-C (1983) Changes in cell shape and actin distribution induced by constant electric fields. Nature 303:61–64

    Google Scholar 

  • Marangos PJ, Zomzely-Neurath C, York C (1975) Immunological studies of a nerve specific protein (NSP). Biochem Biophys 170:289

    Google Scholar 

  • Massa PT, Mugnaini E (1984) Cell-cell junctional interactions and characteristic plasma membrane features of cultured rat glial cells. Neuroscience 2:695–709

    Google Scholar 

  • McCarthy KD, De Vellis J (1980) Preparation of separate astroglial and oligodendroglia cell cultures from rat cerebral tissue. J Cell Biol 85:890–902

    Google Scholar 

  • McGarvey ML, Baron-Van Evercooren A, Kleinman HK, Dubois-Dalcq M (1984) Synthesis and effects of basement membrane components in cultured rat Schwann cells. Dev Biol 104:1–11

    Google Scholar 

  • Mirsky R, Winter J, Abney ER, Pruss RM, Gavrilovic J, Raff MC (1980) Myelin specific proteins and glycolipids in rat Schwann cells and oligodendrocytes in culture. J Cell Biol 84:483–494

    Google Scholar 

  • Mori S, Leblond C (1970) Electron microscopic identification of three classes of oligodendrocytes and a preliminary study of their proliferative capacity in the corpus callosum of young rats. J Comp Neurol 139:1–30

    Google Scholar 

  • Morell P, Toews AD (1984) In vivo metabolism of oligodendroglial lipids. In: Norton WT (ed) Oligodendroglia. Adv Neurochem. Vol 5. Plenum Press, New York, pp 47–86

    Google Scholar 

  • Norton WT (1981) Biochemistry of Myelin. In: Waxman SG, Ritchie JM (eds) Adv Neurobiol Vol 31. Raven Press, New York, pp 93–121

    Google Scholar 

  • Peters A, Palay SL, Webster H deF (1976) The fine structure of the nervous system: the neurons and supporting cells. W.B. Saunders Company, Philadelphia, Chapters VI and VII, pp 181–263

    Google Scholar 

  • Pruss RM (1979) Thy-1 antigen on astrocytes in long term cultures of rat central nervous system. Nature (Lond) 280:688–690

    Google Scholar 

  • Pruss RM, Mirsky R, Raff MC, Thorpe A, Dowding AJ, Anderson BH (1981) All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody. Cell 27:419–428

    Google Scholar 

  • Radice GP (1980) Locomotion and cell-substratum contacts of Xenopus epidermal cells in vitro and in situ. J Cell Sci 44:201–223

    Google Scholar 

  • Raff MC, Mirsky R, Fields KL, Lisak RP, Dorfman SH, Silberberg DH, Gregson NA, Leibowitz S, Kennedy MC (1978) Galactocerebroside is a specific cell-surface antigenic marker for oligodendrocytes in culture. Nature 274:813–816

    Google Scholar 

  • Raff MC, Fields KL, Hakomori S, Mirsky R, Pruss RM, Winter J (1979) Cell-type specific markers for distinguishing and studying neurons and the major classes of glial cells in culture. Brain Res 174:283–308

    Google Scholar 

  • Raff MC, Abney EA, Cohen J, Lindsay R, Noble MJ (1983a) Two types of astrocytes in cultures of developing rat white matter: differences in morphology, surface gangliosides and growth characters. J Neurosci 3:1289–1300

    Google Scholar 

  • Raff MC, Miller RH, Noble M (1983b) A glial progenitor cell that develops in vitro into an astrocyte or an oligodendrocyte depending on culture medium. Nature 303:390–396

    Google Scholar 

  • Raff MC, Williams BP, Miller RH (1984) The in vitro differentiation of a bipotential glial progenitor cell. EMBO J 8:1857–1864

    Google Scholar 

  • Raine CS (1984) Morphology of myelin and myelination. In: Morrell P (eds) Myelin. 2nd Edition. Plenum Press, New York, pp 1–41

    Google Scholar 

  • Ranscht B, Clapshaw PA, 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

    Google Scholar 

  • Raper JA, Bastiani MJ, Goodman CS (1983) Guidance of neuronal growth cones: selective fasciculation in the grasshopper embryo. Cold Spring Harbor Symposia on Quantitative Biology, Vol XLVIII, Mol Neurobiol, pp 587–598

  • Ritchie JM (1984) Physiological basis of conduction in myelinated nerve fibers. In: Morrell P (eds) Myelin. 2nd Edition. Plenum Press, New York, pp 117–141

    Google Scholar 

  • Schmechel DE, Brightman MW, Barker JL (1980) Localization of neuron-specific enolase in mouse spinal cord neurons grown in tissue culture. Brain Res 181:391–400

    Google Scholar 

  • Schnapp BJ, Vale R, Sheetz M, Reese TS (1985) Single microtubules from squid axoplasm support bidirectional movement of organelles. Cell 40:455–462

    Google Scholar 

  • Sternberger NH (1984) Patterns of oligodendrocyte function seen by immunocytochemistry. In: Norton WT (ed) Oligodendroglia. Adv Neurochem. Vol 5. Plenum Press, New York, pp 125–173

    Google Scholar 

  • Sternberger NH, Itoyama Y, Kies MW, Webster H deF (1978) Immunocytochemical method to identify basic protein in myelin-forming oligodendrocytes of newborn rat CNS. J Neurocytol 7:251–263

    Google Scholar 

  • Yahara S, Kishimoto Y, Poduslo JF (1980) High performance liquid chromatography of membrane glycolipids. Assessment of cerebroside on the surface of myelin. In: Sweeland CC (ed) Cell surface glycolipids. Washington, D.C. Am Chem Soc, pp 15–33

    Google Scholar 

  • Wood P, Bunge PR (1984) The biology of oligodendrocyte. In: Norton WT (ed) Oligodendroglia. Adv Neurochem. Vol 5. Plenum Press, New York, pp 1–46

    Google Scholar 

  • Zeller N, Behar T, Dubois-Dalcq M, Lazzarini RA (1986) Timely expression of myelin basic protein gene in rat brain oligodendrocytes cultured in the absence of neurons. J Histochem Cytochem (In press)

  • Zigmond SH (1978) Chemotaxis by polymorphonuclear leukocytes. J Cell Biol 77: 269–287

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Neurobiology, National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    B. Kachar

  2. Laboratory of Molecular Genetics, National Institute of Neurological and Communicative Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    T. Behar & M. Dubois-Dalcq

Authors
  1. B. Kachar
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  2. T. Behar
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  3. M. Dubois-Dalcq
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Kachar, B., Behar, T. & Dubois-Dalcq, M. Cell shape and motility of oligodendrocytes cultured without neurons. Cell Tissue Res. 244, 27–38 (1986). https://doi.org/10.1007/BF00218378

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