Bulk separation and long-term culture of oligodendrocytes from adult pig brain. II. Some biochemical data.
Oligodendroglial proteins labeled with radioactive amino acids were subjected to one- and two-dimensional polyacrylamide electrophoresis. Bands comigrating with myelin proteins, the basic protein (MBP), the proteolipid protein (PLP), and the Wolfgram protein (WP) doublet, were detected by Coomassie Blue staining and by autoradiography. The identity of the MBP and WP in the cellular material is evidenced by immunoblotting with specific antibodies. A comparative study of myelin samples from rat and pig CNS reveals that WP can be detected immunochemically in both species. Different protein patterns, however, are observed. Three protein bands are found with antibodies against the myelin-associated glycoprotein (MAG). The high-molecular-weight component prevails in pig myelin, whereas the medium-molecular-weight component is predominant in rat myelin. Moreover, two protein bands, of molecular weights 35,000 and 33,000 (Ol 1 and Ol 2), are present in high amounts in oligodendroglial particulate material but are not detectable in myelin. These oligodendroglial characteristic proteins are not species-specific, since they are found in preparations of cat oligodendrocytes as well. Activities of cerebroside sulfotransferase (EC 2.8.2.11) are low in freshly isolated cells and increase during the first week of culture. A reverse course of enzyme activities is observed with 2',3'-cyclic nucleotide 3'-phosphohydrolase (EC 3.1.4.37). Values reach a minimum about day 5 in culture and recover their initial values. At day 10 they remain stable until the end of the third week of the culture period.
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002469:Cell Separation; D002478:Cells, Cultured; D007700:Kinetics; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000302:isolation & purification; D009457:Neuroglia / Q000378:metabolism*; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D015238:Sulfotransferases; D013466:Sulfurtransferases / Q000378:metabolism; D013552:Swine
D009419:Nerve Tissue Proteins; D013466:Sulfurtransferases; D015238:Sulfotransferases; C020556:galactosylceramide sulfotransferase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases