Disruption of vimentin intermediate filaments in transgenic mice by expression of a dominant negative mutant desmin subunit.
To investigate putative functions of vimentin intermediate filaments in the context of intact tissues and the developing organism, a construct (pVDV), driven by the vimentin promoter and encoding a truncated desmin subunit, was introduced into the murine germ line. The mutant desmin was assembly-incompetent and capable of disrupting preexisting vimentin filaments in a dominant negative fashion, both in transgenic mouse tissues and in fibroblast cultures derived from these mice. Mutant desmin expression strongly enhanced vimentin turnover. In tissues of some transgenic mouse lines, high level expression of pVDV occurred in 10 to 40% of vimentin-containing cells and, surprisingly, in 1 to 10% of the skeletal and tongue muscle cells. Immunohistochemical staining of muscle tissue showed a diffuse staining pattern instead of the punctated aggregates into which mutant desmin typically accumulates in other cell types. The overexpression of pVDV and the concomitant disruption of the endogenous vimentin filament network and enhanced vimentin turnover in a significant percentage of cells did not cause detectable developmental abnormalities.
Pieper
F R
FR
Department of Biochemistry, University of Nijmegen, The Netherlands.
Raats
J M
JM
Schaart
G
G
Dunia
I
I
van der Kemp
A
A
Benedetti
E L
EL
Ramaekers
F C
FC
Bloemendal
H
H
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D003893:Desmin / Q000096:biosynthesis* / Q000235:genetics*; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000737:chemistry / Q000648:ultrastructure; D051379:Mice; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009928:Organ Specificity; D014746:Vimentin / Q000378:metabolism*
D003893:Desmin; D014746:Vimentin