Abstract
Background. Several examinations have been performed to identify the genes involved in chronic renal failure using 5/6 nephrectomized rats. Recently, many systematic techniques for examining molecular expression have been developed. They might also be effective in elucidating the molecular mechanism of progressive renal failure. In this study, digital expression pro filing was carried out to construct a subtractive mRNA expression database for the 5/6 nephrectomized kidney. Methods. One thousand clones were randomly sequenced from 5/6 nephrectomized and sham-operated rat kidney cDNA libraries, respectively, and defined by BLAST search. In silico subtractive analysis was performed to search for genes up- or down-regulated in the 5/6 nephrectomized kidney. Results. The growth factor-related mRNAs and the mRNAs encoding cytoskeletal or membrane proteins were up-regulated, but the transporter-related mRNAs were down-regulated in the 5/6 nephrectomized kidney database. In silico subtraction revealed that 63 mRNAs were increased and 59 were decreased in the 5/6 nephrectomized kidney. To confirm whether the in silico subtractive database reflected the actual expression of mRNA or protein, 12 known genes were examined by Northern blotting or immunoblotting, respectively. The actual expression of the 12 genes was comparable with the results of in silico subtraction. In addition, we successfully isolated five unknown genes, two up-regulated and three down-regulated in the 5/6 nephrectomized kidney. Conclusion. We constructed a subtractive mRNA expression database for 5/6 nephrectomized kidney, which reflects the actual alterations in mRNA expression after subtotal nephrectomy. This database may be useful for elucidation of the molecular mechanism of progressive renal failure.
| Original language | English |
|---|---|
| Pages (from-to) | 29-45 |
| Number of pages | 17 |
| Journal | Kidney International |
| Volume | 66 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Jul 2004 |
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All Science Journal Classification (ASJC) codes
- Nephrology
Cite this
Gene expression variance based on random sequencing in rat remnant kidney. / Horiba, Naoshi; Masuda, Satohiro; Takeuchi, Ayako; Saito, Hideyuki; Okuda, Masahiro; Inui, Ken Ichi.
In: Kidney International, Vol. 66, No. 1, 07.2004, p. 29-45.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Gene expression variance based on random sequencing in rat remnant kidney
AU - Horiba, Naoshi
AU - Masuda, Satohiro
AU - Takeuchi, Ayako
AU - Saito, Hideyuki
AU - Okuda, Masahiro
AU - Inui, Ken Ichi
PY - 2004/7
Y1 - 2004/7
N2 - Background. Several examinations have been performed to identify the genes involved in chronic renal failure using 5/6 nephrectomized rats. Recently, many systematic techniques for examining molecular expression have been developed. They might also be effective in elucidating the molecular mechanism of progressive renal failure. In this study, digital expression pro filing was carried out to construct a subtractive mRNA expression database for the 5/6 nephrectomized kidney. Methods. One thousand clones were randomly sequenced from 5/6 nephrectomized and sham-operated rat kidney cDNA libraries, respectively, and defined by BLAST search. In silico subtractive analysis was performed to search for genes up- or down-regulated in the 5/6 nephrectomized kidney. Results. The growth factor-related mRNAs and the mRNAs encoding cytoskeletal or membrane proteins were up-regulated, but the transporter-related mRNAs were down-regulated in the 5/6 nephrectomized kidney database. In silico subtraction revealed that 63 mRNAs were increased and 59 were decreased in the 5/6 nephrectomized kidney. To confirm whether the in silico subtractive database reflected the actual expression of mRNA or protein, 12 known genes were examined by Northern blotting or immunoblotting, respectively. The actual expression of the 12 genes was comparable with the results of in silico subtraction. In addition, we successfully isolated five unknown genes, two up-regulated and three down-regulated in the 5/6 nephrectomized kidney. Conclusion. We constructed a subtractive mRNA expression database for 5/6 nephrectomized kidney, which reflects the actual alterations in mRNA expression after subtotal nephrectomy. This database may be useful for elucidation of the molecular mechanism of progressive renal failure.
AB - Background. Several examinations have been performed to identify the genes involved in chronic renal failure using 5/6 nephrectomized rats. Recently, many systematic techniques for examining molecular expression have been developed. They might also be effective in elucidating the molecular mechanism of progressive renal failure. In this study, digital expression pro filing was carried out to construct a subtractive mRNA expression database for the 5/6 nephrectomized kidney. Methods. One thousand clones were randomly sequenced from 5/6 nephrectomized and sham-operated rat kidney cDNA libraries, respectively, and defined by BLAST search. In silico subtractive analysis was performed to search for genes up- or down-regulated in the 5/6 nephrectomized kidney. Results. The growth factor-related mRNAs and the mRNAs encoding cytoskeletal or membrane proteins were up-regulated, but the transporter-related mRNAs were down-regulated in the 5/6 nephrectomized kidney database. In silico subtraction revealed that 63 mRNAs were increased and 59 were decreased in the 5/6 nephrectomized kidney. To confirm whether the in silico subtractive database reflected the actual expression of mRNA or protein, 12 known genes were examined by Northern blotting or immunoblotting, respectively. The actual expression of the 12 genes was comparable with the results of in silico subtraction. In addition, we successfully isolated five unknown genes, two up-regulated and three down-regulated in the 5/6 nephrectomized kidney. Conclusion. We constructed a subtractive mRNA expression database for 5/6 nephrectomized kidney, which reflects the actual alterations in mRNA expression after subtotal nephrectomy. This database may be useful for elucidation of the molecular mechanism of progressive renal failure.
UR - http://www.scopus.com/inward/record.url?scp=3042802208&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=3042802208&partnerID=8YFLogxK
U2 - 10.1111/j.1523-1755.2004.00704.x
DO - 10.1111/j.1523-1755.2004.00704.x
M3 - Article
C2 - 15200410
AN - SCOPUS:3042802208
VL - 66
SP - 29
EP - 45
JO - Kidney International
JF - Kidney International
SN - 0085-2538
IS - 1
ER -