Expression and localization of myelin basic protein in oligodendrocytes and transfected fibroblasts.
Myelin basic protein (MBP) is a major structural component of myelin. It is expressed exclusively in myelinating glia (oligodendrocytes in the CNS and Schwann cells in the PNS) and is localized to the cytoplasmic surface of the plasma membrane and myelin membrane produced by these cells. The work described here concerns the mechanism of plasma membrane localization of MBP in myelinating glial cells and whether it involves differentiated functions specific to these cells or general functions of plasma membrane assembly common to all cells. To this end, the subcellular localization of endogenous MBP in mouse oligodendrocytes was compared with that of transiently expressed MBP in monkey fibroblasts (Cos-1 cells) transfected with an MBP expression vector containing cDNA for rat 14K MBP. The steady-state levels of MBP-specific RNA and of MBP polypeptide expressed in the transfected fibroblasts were comparable to the levels expressed in oligodendrocytes in primary culture. MBP localization was analyzed in whole cells by immunofluorescence and in specific intracellular compartments by subcellular fractionation. The results show that MBP expressed in wild-type oligodendrocytes is localized to the plasma membrane. In contrast, MBP expressed in transfected fibroblasts appears dispersed in the cytoplasm and is distributed uniformly among the various subcellular fractions.(ABSTRACT TRUNCATED AT 250 WORDS)
Barbarese
E
E
Department of Neurology, University of Connecticut Health Center, Farmington 06032.
Barry
C
C
Chou
C H
CH
Goldstein
D J
DJ
Nakos
G A
GA
Hyde-DeRuyscher
R
R
Scheld
K
K
Carson
J H
JH
D000818:Animals; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D000882:Haplorhini; D051379:Mice; D008809:Mice, Inbred C3H; D004676:Myelin Basic Protein / Q000235:genetics / Q000378:metabolism*; D009457:Neuroglia / Q000378:metabolism*; D009836:Oligodendroglia / Q000378:metabolism*; D012313:RNA / Q000235:genetics; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D004676:Myelin Basic Protein; D012313:RNA; D004247:DNA