CD2AP expression in a renal ischemia/reperfusion injury model and analysis of its related genes using overexpression and RNA interference technique.
BACKGROUND
CD2-associated protein (CD2AP) is a ubiquitously expressed 80-kDa intracellular protein, and has been speculated to act as an intracellular signaling pathway between plasma membrane proteins and cytoskeleton proteins. CD2AP expression has been reported in both the glomerulus and tubular epithelium in the kidney, and CD2AP knockout mice exhibit congenital nephrotic syndrome. However, the precise properties and its role in the renal tubules have not been clarified.


METHODS
We used an established rat model of ischemic/reperfusion renal injury (IRI) to examine the expression of CD2AP by real-time PCR, Western blotting, and immunohistochemistry. We also investigated the expression of genes related to apoptosis and cell proliferation in mouse collecting duct-derived cells (M1 cells) transfected with full-length of CD2AP cDNA or short interfering RNA.


RESULTS
CD2AP mRNA and protein expression had significantly increased in the IRI kidney. Real-time PCR indicated that expression of genes regulating apoptosis, such as B-Raf and Caspase-12, and genes regulating cell proliferation factors, CDC2, was decreased in CD2AP-overexpressing M1 cells and increased in CD2AP-interfered M1 cells.


CONCLUSIONS
These results suggest that CD2AP expression was increased following renal ischemia and that CD2AP may be related to the process of cell repair and/or cell differentiation following injury.
Nishimura
Sayoko
S
Department of Medicine IV, Tokyo Women's Medical University, Japan.
Yoshida
Takumi
T
Sugiura
Hidekazu
H
Shirota
Satsuki
S
Tsuchiya
Ken
K
Nihei
Hiroshi
H
D033942:Adaptor Proteins, Vesicular Transport / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D007674:Kidney Diseases / Q000235:genetics* / Q000378:metabolism; D007685:Kidney Tubules, Collecting / Q000378:metabolism; D008297:Male; D034622:RNA Interference; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000235:genetics* / Q000378:metabolism
D033942:Adaptor Proteins, Vesicular Transport