Polypeptides arising from neutral in vitro proteolysis of myelin basic protein (MBP) of human brain were evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. At pH 7 a marked breakdown of MBP resulted in the formation of 8-12 polypeptides ranging from 6 to 17 kd in molecular weight. As neutral proteolytic activity was not eliminated by either gel filtration or cation-exchange chromatography acid-soluble protease(s) involved probably have a size and electric charge similar to that of MBP. The enzymatic nature of neutral proteolysis was ascertained by heat inactivation and inhibition by alpha 2-macroglobulin. Incomplete inhibition of proteolysis and the failure of small peptides (less than 6 kd) to show up on electrophoresis seem to suggest that MBP was degraded by exopeptic proteases as well. Acid extracts of purified myelin yielded polypeptides similar to those of MBP of delipidated white matter. The results are consistent with a sequential limited proteolysis of MBP by neutral proteases probably associated with myelin and possibly related to the in situ catabolism of MBP in man.
Alcohol was administered in a vitamin-supplemented liquid diet (Stardit) to Wistar rats 4 weeks before mating and during pregnancy. Of the daily calories 38--40% were supplied by alcohol and by isocaloric sucrose in the controls. Offspring of alcohol drinking dams showed a reduced birth weight and gained weight less rapidly than the controls. On day 3, 4, 7, 12, 17 and 21, experimental animals and controls were perfused for histological and electron microscopic investigations. Morphometric analysis of the cerebella (Fol. 5 and Fol. 8) showed no difference in the number of Purkinje cells. However, on day 7 the Purkinje cell nuclei of experimental pups were significantly smaller. This difference disappeared at day 12. Electron microscopic investigations in the 4, 7, and 12 days old experimental animals revealed a delayed cytoplasmic maturation of Purkinje cells which mainly involved the rough endoplasmic reticulum. At 17 days, there was no difference between the two groups. These data are discussed in relation to alcohol-induced metabolic changes in the brain.
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