Co-clustering of galactosylceramide and membrane proteins in oligodendrocyte membranes on interaction with polyvalent carbohydrate and prevention by an intact cytoskeleton.
We have shown previously that addition of liposomes containing the two major glycosphingolipids of myelin, galactosylceramide (GalC) and cerebroside sulfate (CBS), to cultured oligodendrocytes (OLs) caused clustering of GalC on the extracellular surface and myelin basic protein (MBP) on the cytosolic surface to the same membrane domains. It also caused depolymerization of actin microfilaments and microtubules, indicating that interaction of the liposomes with the OL surface induces transmembrane signal transmission. We show that a multivalent form of galactose conjugated to bovine serum albumin has a similar effect as the multivalent GalC/CBS-containing liposomes. Because GalC and CBS can interact with each other across apposed membranes and because anti-GalC and anti-CBS antibodies also cause redistribution of GalC/CBS and depolymerization of microtubules, we believe that the multivalent carbohydrate interacts with GalC and CBS in the OL membrane. Several myelin-specific transmembrane proteins could be involved in this transmembrane signal transmission from GalC/CBS. We looked at co-clustering of several myelin constituents by confocal microscopy to determine if they are located in or redistribute to GalC/MBP-containing domains. Myelin oligodendrocyte glycoprotein (MOG), proteolipid protein (PLP), MAPK, and some phosphotyrosine-containing proteins were found to co-cluster with GalC and MBP, but myelin-associated glycoprotein (MAG) and phosphatidylinositol-4,5-bisphosphate (PIP(2)) did not. These results suggest that MOG and PLP, but probably not MAG, are possible candidates for transmembrane transmission of the signal received by GalC/CBS. To determine if depolymerization of actin microfilaments was required for co-clustering, or was secondary to clustering, we stabilized F-actin with jasplakinolide. This also prevented depolymerization of the microtubules and prevented clustering of all constituents, including GalC. The prevention of clustering or redistribution of these glycolipids and proteins by an intact cytoskeleton is consistent with the picket fence model.
Boggs
Joan M
JM
Department of Structural Biology and Biochemistry, Hospital for Sick Children, Toronto, ON, Canada. jmboggs@sickkids.ca
Wang
Huimin
H
D000199:Actins / Q000378:metabolism / Q000502:physiology; D000818:Animals; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D005690:Galactose / Q000737:chemistry / Q000378:metabolism*; D005699:Galactosylceramides / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D009836:Oligodendroglia / Q000378:metabolism*; D011487:Protein Conformation; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000502:physiology
D000199:Actins; D002241:Carbohydrates; D005699:Galactosylceramides; D008081:Liposomes; D008565:Membrane Proteins; D005690:Galactose