Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD.
MYELINATION IN RAT BRAIN: METHOD OF MYELIN ISOLATION1
Version of Record online: 4 OCT 2006
Journal of Neurochemistry
Volume 21, Issue 4, pages 749–757, October 1973
How to Cite
Norton, W. T. and Poduslo, S. E. (1973), MYELINATION IN RAT BRAIN: METHOD OF MYELIN ISOLATION. Journal of Neurochemistry, 21: 749–757. doi: 10.1111/j.1471-4159.1973.tb07519.x
This study was supported by U.S. Public Health Service grants NS-02476, NS-01006 and NS-03356.
- Issue online: 4 OCT 2006
- Version of Record online: 4 OCT 2006
- Received 29 December 1972. Accepted 17 April 1973
Abstract— A procedure is described for the preparation from rat brain of myelin having the same degree of purity at all ages. Such a procedure is essential for the study of myelin composition during development. Microsomal contamination was successfully eliminated by adjusting the method to maintain a constant amount of brain per unit volume in the initial density gradient step, and by including two osmotic shocks and two low-speed centrifugation steps. Myelin prepared in this way from animals ranging from 15 days to 14months of age had a total ATPase activity of 0.3-2.0 μmol of Pi.h−1.mg−1 dry wt of myelin, representing 0.1-1.2 per cent recovery of the total homogenate activity; a Na+, K+- ATPase activity of 0.1-1.6 μfnol of Pi.h−1.mg−1 dry wt, representing 0.04-1.5 per cent recovery; a nucleic acid content of 0.2-0.7 per cent of myelin dry wt, representing 0.2-2.0 per cent recovery; and a ganglioside NANA content of 0.04-0.07 per cent of myelin dry wt. representing 0.2-4.6 per cent recovery. The myelin prepared from 20-day animals had the highest content of the first three constituents; otherwise the values of the four constituents were relatively constant per unit weight of myelin. The amounts of nucleic acid and ganglioside recovered in the myelin fractions increased with increasing age and myelin yield.