Rumpshaker-like proteolipid protein (PLP) ratio in a mouse model with unperturbed structural and functional integrity of the myelin sheath and axons in the central nervous system.
The gene plp on the X chromosome encodes the isoforms proteolipid protein (PLP) and DM(20), two dominant integral membrane proteins of central nervous system (CNS) myelin. DM(20) results from the activation of the cryptic splice site in exon III of the PLP gene. We inserted a sense-orientated loxP flanked neomycin-gene into intron III of the plp sequence, using homologous recombination in embryonic stem cells and generated the homozygous neoS mouse line. Unlike the previously described complete PLP/DM(20) ablation (plp(-/-)), which has been obtained by introducing a neo-gene in antisense-orientation in the same position of intron III, the plp expression surprisingly revealed reduced mRNA levels. The PLP isoform was reduced to 50%, but DM(20) expression was unaffected. This protein pattern resembles the expression profile of the PLP isoforms in the natural occurring rumpshaker mutant. Electron microscopic examination revealed a normal compaction of CNS-myelin and maintenance of axon integrity. PLP expression levels of the wt control were recovered by Cre excision of the neo-selection gene after intercrossing neoS mice and oligodendrocyte-specific Cre-mice. These data strongly hint at different functions of intron III in PLP/DM(20)-specific splicing and mRNA stability. Furthermore evidence is provided for functionally affected translation products of the PLP gene in the rumpshaker mutant, whereas no PLP-isoform occur in plp(-/-) mice generated by introducing a selectable marker into intron III in antisense orientation.
Uschkureit
T
T
Laboratory for Molecular Neuroscience, University of Cologne, Cologne, Germany.
Spörkel
O
O
Büssow
H
H
Stoffel
W
W
D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001369:Axons / Q000378:metabolism*; D002490:Central Nervous System / Q000002:abnormalities* / Q000254:growth & development / Q000648:ultrastructure; D004195:Disease Models, Animal; D018507:Gene Expression Regulation, Developmental; D018390:Gene Targeting / Q000379:methods; D007438:Introns / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout / Q000002:abnormalities / Q000235:genetics / Q000254:growth & development; D008818:Mice, Neurologic Mutants / Q000002:abnormalities / Q000235:genetics* / Q000254:growth & development; D018991:Myelin Proteolipid Protein / Q000235:genetics* / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism* / Q000648:ultrastructure; D009355:Neomycin; D009419:Nerve Tissue Proteins; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics
D018991:Myelin Proteolipid Protein; D009419:Nerve Tissue Proteins; C047516:Plp1 protein, mouse; D012333:RNA, Messenger; D009355:Neomycin