Expression of human mid-sized neurofilament subunit in transgenic mice.
We have created transgenic mice which carry and express the gene encoding the human NF(M) subunit. RNAase protection assays reveal that the transgene is abundantly expressed in CNS and PNS but also, at very low levels in some non-neural tissues as well. Although the neurospecificity of transgene transcription was not absolute, we are able to detect the protein only in neurons with immunocytochemical techniques. Glial and endothelial cells do not contain immunoreactive materials. Interesting subtle differences in the relative level of the human transgene encoded and endogenous murine encoded NF(M) proteins were noted in different regions of the brain. Similar differences were found in the levels of transgene and endogenous gene mRNA suggesting that these differences may be traceable to differences in RNA transcription or stability. Our data demonstrate, within the sensitivity of the immunocytochemical techniques we used, that the human NF(M) protein is present only in the neurons of the transgenic mice and that it is present in the same neurons as the endogenous NF(M). Furthermore, immunoelectron-microscopic examination of isolated neurofilaments shows that the human NF(M) coassembles with the endogenous NF(M) during filament formation. Thus, although the human NF(M) possesses a much larger multiphosphorylation site in its carboxy terminus, it seems to be the functionally equivalent to the mouse protein, even in the murine neuron.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D001923:Brain Chemistry; D004247:DNA / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D016253:Microscopy, Immunoelectron; D009474:Neurons / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis; D012260:Ribonucleases / Q000096:biosynthesis; D013116:Spinal Cord / Q000378:metabolism
D000911:Antibodies, Monoclonal; D012313:RNA; D004247:DNA; D012260:Ribonucleases