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The Journal of Cell Biology, Vol 111, 1543-1557, Copyright © 1990 by The Rockefeller University Press
ARTICLES |
MJ Monteiro, PN Hoffman, JD Gearhart and DW Cleveland
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
We have generated transgenic mice containing additional copies of the murine NF-L gene in order to examine the consequences of neurofilament- L overexpression on axonal morphology. Founder mice were constructed to carry a transgene in which the presumptive 5' promoter sequences of NF- L were replaced with the strong murine sarcoma virus long terminal repeat promoter. The transgenes were expressed prominently in several tissues, including skeletal muscle and kidney where NF-L accumulated to approximately 2% of cell protein. This was not accompanied by an overt phenotype, except that expression in lens led to cataract formation. In the brains of these animals, transgene RNA levels exceeded the endogenous NF-L RNAs by up to 20-fold, although no additional protein accumulated, indicating posttranscriptional regulation of NF-L expression. However, in peripheral neurons transgene RNA was approximately fourfold higher than endogenous NF-L mRNAs, and a corresponding increase in NF-L subunits was found in axons arising from these neurons. Myelinated nerve fibers of transgenic animals contained increased numbers of NFs, assembled predominantly of NF-L. This was reflected in an increase in the density of axonal NFs; axonal caliber was not affected.
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