Abstract
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, ε-sarcoglycan, ribosomal protein S3a, G-proteinγ10 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.
| Original language | English |
|---|---|
| Pages (from-to) | 549-558 |
| Number of pages | 10 |
| Journal | Kidney International |
| Volume | 56 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 1999 |
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Keywords
- Gene expression
- Glomerular hyperfiltration
- Nephrectomy
- Remnant kidney
- Subtractive screening
ASJC Scopus subject areas
- Nephrology
Cite this
Screening for genes up-regulated in 5/6 nephrectomized mouse kidney. / Zhang, Hong; Wada, Jun; Kanwar, Yashpal S.; Tsuchiyama, Yoshinori; Hiragushi, Keita; Hida, Kazuyuki; Shikata, Kenichi; Makino, Hirofumi.
In: Kidney International, Vol. 56, No. 2, 1999, p. 549-558.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Screening for genes up-regulated in 5/6 nephrectomized mouse kidney
AU - Zhang, Hong
AU - Wada, Jun
AU - Kanwar, Yashpal S.
AU - Tsuchiyama, Yoshinori
AU - Hiragushi, Keita
AU - Hida, Kazuyuki
AU - Shikata, Kenichi
AU - Makino, Hirofumi
PY - 1999
Y1 - 1999
N2 - 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, ε-sarcoglycan, ribosomal protein S3a, G-proteinγ10 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.
AB - 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, ε-sarcoglycan, ribosomal protein S3a, G-proteinγ10 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.
KW - Gene expression
KW - Glomerular hyperfiltration
KW - Nephrectomy
KW - Remnant kidney
KW - Subtractive screening
UR - http://www.scopus.com/inward/record.url?scp=0032782374&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032782374&partnerID=8YFLogxK
U2 - 10.1046/j.1523-1755.1999.00561.x
DO - 10.1046/j.1523-1755.1999.00561.x
M3 - Article
C2 - 10432394
AN - SCOPUS:0032782374
VL - 56
SP - 549
EP - 558
JO - Kidney International
JF - Kidney International
SN - 0085-2538
IS - 2
ER -