Screening for genes up-regulated in 5/6 nephrectomized mouse kidney.
BACKGROUND
In diabetic and nondiabetic renal diseases, glomerular hyperfiltration is believed to play a central role in the subsequent progression of glomerulosclerosis and interstitial renal scarring. To identify genes involved in the process of hyperfiltration and hypertrophy, a polymerase chain reaction (PCR)-based subtraction method, that is, representational difference analysis of cDNA (cDNA-RDA), was employed.


METHODS
Ten-week-old ICR mice were 5/6 nephrectomized and sham operated. After two weeks, mRNAs were isolated from control and remnant kidneys and were subjected to the cDNA-RDA procedure.


RESULTS
We identified 10 known and 9 novel genes. Among 19 clones, 12 clones (8 known and 4 novel) showed 1.5- to 6-fold up-regulation by Northern blot analyses. The remaining seven clones were rarely expressed genes and were barely detected by Northern blot analyses, and their up-regulated expression was confirmed by Southern blot analysis using the PCR-amplified representative amplicons. The known genes included kidney androgen-regulated protein, major urinary protein, lysozyme M, metalloproteinase-3 tissue inhibitor, chaperonin 10, cytochrome oxidase I, epsilon-sarcoglycan, ribosomal protein S3a, G-proteingamma10 subunit, and splicing factor 9G8. All of the isolated known genes have not been reported to be up-regulated in the nephrectomized mouse kidney and suggest the possible role of androgen action, mitochondrial functions, matrix metabolism, cell-matrix interactions, and intracellular signaling events in the initiation of the progressive renal injury of the remnant kidney. Furthermore, cDNA-RDA facilitates the discovery of novel genes, including two kidney-specific genes.


CONCLUSIONS
The isolated known and novel genes may be involved in the pathobiological process of initial hyperfiltration and hypertrophy of remnant kidney.
Zhang
H
H
Department of Medicine III, Okayama University Medical School, Japan.
Wada
J
J
Kanwar
Y S
YS
Tsuchiyama
Y
Y
Hiragushi
K
K
Hida
K
K
Shikata
K
K
Makino
H
H
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002921:Cicatrix / Q000473:pathology; D018076:DNA, Complementary / Q000032:analysis; D003921:Diabetes Mellitus, Experimental / Q000235:genetics; D003928:Diabetic Nephropathies / Q000235:genetics; D015870:Gene Expression / Q000502:physiology; D005820:Genetic Testing; D005923:Glomerulosclerosis, Focal Segmental / Q000235:genetics*; D006984:Hypertrophy; D007668:Kidney / Q000473:pathology / Q000502:physiology* / Q000601:surgery; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009392:Nephrectomy; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis
D018076:DNA, Complementary; D012333:RNA, Messenger