Oligodendroglia development in cell culture as monitored with a monoclonal antibody

Abstract

A new marker for young oligodendrocytes has been identified by a monoclonal antibody (mOg-1, IgM isotype) prepared from cerebellar plasma membrane stimulated mouse lymphocytes. mOg-1 reactive cells in the mouse cerebellum first appear at day 19 of gestation. Future white matter layers of fixed sections of neonatal rat cerebellum were labeled with mOg-1. Although EM analysis has shown cell-surface binding by presumptive oligodendroglia in neonatal cerebellum, the antibody does not bind to compact myelin. In cell cultures prepared from L-d-old mice, 1.1% of the cells bound mOg-1 after 3 d in culture, but up to 5% of the cells bound mOg-1 after 2 weeks in culture. Of these same Og-l-positive cells, 69% bound anti-gabtctocerebroside and 65% bound anti-myelin basic protein. After a week in culture Og-l-positive cells often produced lamellar sheets extending a millimeter over the polylysine substratum in the absence of normal myelin formation. mOg-1 recognizes a cell-surface determinant distinct from well-characterized oligodendroglial molecules (galactocerebroside, sulfatide and myelin basic protein) that is expressed early in oligodendrocyte development. The antibody has been used to follow the maturation of oligodendrocytes in cultures of both normal and jimpy mouse cerebellum.The nervous system is a complex network derived from multiple associations between different cell types. The limited number of distinct cell types and the availability of neurological mutants with defective cerebellar histogenesis make the mouse cerebellum an attractive model for studying cell-cell interactions.The development of antibodies toward neural cell-surface molecules could be instrumental in establishing the role of specific determinants in histogenesis. We have used the hybridoma technology of Milstein (1975, 1976) In this report, we describe a monoclonal antibody (mOg-1) reactive with the surface of young oligodendrocytes, prior to the Received Dec. 19, 1985; revised Mar. 4, 1986; accepted Mar. 4, 1986. We woula like to thank Dr. Robert Lasher for his helpful discussions and S.Hat&e for her expert assistance. We thank Drs. Dahl, De.Vellis, Evans, Habig, Qua&s, Ranscht, and Rapport for their generous girt of antisera and tetanus toxin used in these studies. Some of these studies were submitted in partial fulfillment of the requirementi for the Doctor of Philosophy degree from the University of Colorado (1983). This work was su~ucwted in nart bv USPHS Grants NS-098 18and T32-bM-07342.

Immunocytochemical localization by electron microscopy of 2'3'-cyclic nucleotide 3'-phosphodiesterase in developing oligodendrocytes of normal and mutant brain

“…At this age we observed many immunostained oligodendrocytes, often bearing remarkably intricate, fine processes or lacelike filopodia reminiscent of cells grown in culture (Collins and Seeds, 1986;Kachar et al, 1986;Knapp et al, 1987). This is significant because the question has been raised as to whether or not the intricate network of fingerlike projections observed in vitro is actually present in vivo.…”

“…These have provided interesting and valuable data on progenitor cells and their differentiation into oligodendrocytes or astrocytes and on early development (Barbarese et al, 1983;Raffet al, 1983;Hirayama et al, 1984;Collins and Seeds, 1986;Goldman et al, 1986). Unlike anti-galactocerebroside, anti-CNP can be used for both light-and electron-microscopic studies as an integrative tool with which to evaluate in vivo the validity of interpretations derived from in vitro studies.…”

Characterization of 125I-tissue plasminogen activator binding to cerebellar granule neurons.

“…The coverslips were rapidly rinsed and fixed for 10 min in 4% pamformaldehyde, rinsed, after which some coverslips were treated with -20"C methanol for 5 min, then incubated for 20 min with rabbit anti-glial fibrillary acidic protein (11), and rinsed. All coverslips were incubated for 20 min with fluorescein-conjugated goat anti-rabbit IgG as described previously (10), then rinsed, air dried and mounted for autoradiography with NTB2 emulsion as described above. After development and processing, the coverslips were viewed with an epifluorescence microscope (Carl Zeiss, Inc., Thornwood, NY) using a 63 x planapochromat objective.…”

Myelin acquisition in the central nervous system of the mouse revealed by an MBP-Lac Z transgene

“…As a result, the cellular processes and many of the molecules involved in myelin elaboration have been identified and extensively characterized. Despite the fact that oligodendrocyte precursors can differentiate in vitro into cells that express many myelin-specific components (Zeller et al, 1985;Collins and Seeds, 1986;Dubois-Dalcq et al, 1986), various factors including those derived from neurons and astrocytes can exert a strong influence on this process (David et al, 1984;Shanker et al, 1987;Chen and DeVries, 1989;Giulian et al, 199 1). Therefore, myelin elaboration in vivo is likely regulated by a number of diffusible and membraneassociated molecules present in the local environment of oligodendrocytes.…”