Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Cell shape and motility of oligodendrocytes cultured without neurons

  • 78 Accesses

  • 45 Citations

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.

This is a preview of subscription content, log in to check access.

Abbreviations

(CNS):

Central nervous system

(DIC):

Differential interference contrast

(GC):

Galactocerebroside

(GFA) protein:

Glial fibrillary acidic

(NSE):

Neuron-specific enolase

References

  1. 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

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

  3. 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

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

  5. 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

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

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

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

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

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

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

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

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

  14. 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

  15. 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

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

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

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

  19. 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

  20. 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

  21. 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

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

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

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

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

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

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

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

  29. 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

  30. 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

  31. 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

  32. 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

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

  34. 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

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

  36. 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

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

  38. 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

  39. 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

  40. 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

  41. 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

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

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

  44. 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

  45. 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

  46. 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

  47. 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

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

  49. 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

  50. 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

  51. 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

  52. 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

  53. 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)

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

Download references

Author information

Correspondence to B. Kachar.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

Key words

  • Oligodendroglial cells (rat)
  • Myelin
  • Cell movements
  • Cell culture
  • CNS
  • Cell differentiation