Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft

M. Michimata, W. Wang, S. Fujita, H. Mizutani, K. Fujimori, S. Satomi, M. Ohta, S. Ito, T. Kimura, T. Araki, Y. Imai, M. Matsubara

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Abstract

Background. The underlying mechanisms of renal transplant dysfunction are poorly understood. There is little information on tubular function in kidney grafts. The cDNAs encoding kidney-specific cell surface proteins required for renal reabsorption of sodium (sodium cotransporter in thick ascending limb of Henle, rBSC1) and water (apical water channel in collecting duct, AQP2) have been recently identified. Since transcripts of these proteins are up-regulated in dehydration in association with maximal concentration of urine, we examined urinary concentrating ability and expression levels of mRNA of these proteins in kidney isografts. Methods. Male Sprague-Dawley rats underwent syngeneic renal transplantation or unilateral nephrectomy (UNX) and were deprived of water for 24 hours at six weeks after the operation when histological and functional compensation of the intact kidney was complete. Blood and urinary samples were collected before and after dehydration. The amount of rBSC1 or AQP2 mRNA was measured using competitive polymerase chain reaction (PCR) by inducing a point mutation at the middle of PCR product for rBSC1 or by deleting 180 bp from 780 bp PCR product for AQP2, respectively. The protein expression was examined by Western blot analysis. Results. Both groups of rats demonstrated the same levels of compensatory renal hypertrophy (∼60% weight increase) and plasma creatinine values. Histological examination revealed enlarged glomeruli and tubules, but no findings of ischemic damage, such as tubular atrophy or interstitial changes. Urinary concentration was noted in the UNX rats but not in rats with kidney grafts. Competitive PCR demonstrated that dehydration did not increase rBSC1 and AQP2 transcripts in rats with kidney transplantation. Immunoblot analysis confirmed that the marked increase of both rBSC1 and AQP2 proteins was noted only in the remnant kidney of dehydrated rats.

Original languageEnglish
Pages (from-to)672-679
Number of pages8
JournalKidney International
Volume60
Issue number2
DOIs
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Transplants
Kidney
Dehydration
Aquaporin 2
Polymerase Chain Reaction
Kidney Transplantation
Sodium
Isogeneic Transplantation
Isografts
Messenger RNA
Proteins
Water
Nephrectomy
Point Mutation
Hypertrophy
Atrophy
Sprague Dawley Rats
Creatinine
Membrane Proteins
Extremities

Cite this

Michimata, M., Wang, W., Fujita, S., Mizutani, H., Fujimori, K., Satomi, S., ... Matsubara, M. (2001). Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft. Kidney International, 60(2), 672-679. https://doi.org/10.1046/j.1523-1755.2001.060002672.x
Michimata, M. ; Wang, W. ; Fujita, S. ; Mizutani, H. ; Fujimori, K. ; Satomi, S. ; Ohta, M. ; Ito, S. ; Kimura, T. ; Araki, T. ; Imai, Y. ; Matsubara, M. / Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft. In: Kidney International. 2001 ; Vol. 60, No. 2. pp. 672-679.
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abstract = "Background. The underlying mechanisms of renal transplant dysfunction are poorly understood. There is little information on tubular function in kidney grafts. The cDNAs encoding kidney-specific cell surface proteins required for renal reabsorption of sodium (sodium cotransporter in thick ascending limb of Henle, rBSC1) and water (apical water channel in collecting duct, AQP2) have been recently identified. Since transcripts of these proteins are up-regulated in dehydration in association with maximal concentration of urine, we examined urinary concentrating ability and expression levels of mRNA of these proteins in kidney isografts. Methods. Male Sprague-Dawley rats underwent syngeneic renal transplantation or unilateral nephrectomy (UNX) and were deprived of water for 24 hours at six weeks after the operation when histological and functional compensation of the intact kidney was complete. Blood and urinary samples were collected before and after dehydration. The amount of rBSC1 or AQP2 mRNA was measured using competitive polymerase chain reaction (PCR) by inducing a point mutation at the middle of PCR product for rBSC1 or by deleting 180 bp from 780 bp PCR product for AQP2, respectively. The protein expression was examined by Western blot analysis. Results. Both groups of rats demonstrated the same levels of compensatory renal hypertrophy (∼60{\%} weight increase) and plasma creatinine values. Histological examination revealed enlarged glomeruli and tubules, but no findings of ischemic damage, such as tubular atrophy or interstitial changes. Urinary concentration was noted in the UNX rats but not in rats with kidney grafts. Competitive PCR demonstrated that dehydration did not increase rBSC1 and AQP2 transcripts in rats with kidney transplantation. Immunoblot analysis confirmed that the marked increase of both rBSC1 and AQP2 proteins was noted only in the remnant kidney of dehydrated rats.",
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Michimata, M, Wang, W, Fujita, S, Mizutani, H, Fujimori, K, Satomi, S, Ohta, M, Ito, S, Kimura, T, Araki, T, Imai, Y & Matsubara, M 2001, 'Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft' Kidney International, vol. 60, no. 2, pp. 672-679. https://doi.org/10.1046/j.1523-1755.2001.060002672.x

Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft. / Michimata, M.; Wang, W.; Fujita, S.; Mizutani, H.; Fujimori, K.; Satomi, S.; Ohta, M.; Ito, S.; Kimura, T.; Araki, T.; Imai, Y.; Matsubara, M.

In: Kidney International, Vol. 60, No. 2, 2001, p. 672-679.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Limited urinary concentration and damaged tubules in rats with a syngeneic kidney graft

AU - Michimata, M.

AU - Wang, W.

AU - Fujita, S.

AU - Mizutani, H.

AU - Fujimori, K.

AU - Satomi, S.

AU - Ohta, M.

AU - Ito, S.

AU - Kimura, T.

AU - Araki, T.

AU - Imai, Y.

AU - Matsubara, M.

PY - 2001

Y1 - 2001

N2 - Background. The underlying mechanisms of renal transplant dysfunction are poorly understood. There is little information on tubular function in kidney grafts. The cDNAs encoding kidney-specific cell surface proteins required for renal reabsorption of sodium (sodium cotransporter in thick ascending limb of Henle, rBSC1) and water (apical water channel in collecting duct, AQP2) have been recently identified. Since transcripts of these proteins are up-regulated in dehydration in association with maximal concentration of urine, we examined urinary concentrating ability and expression levels of mRNA of these proteins in kidney isografts. Methods. Male Sprague-Dawley rats underwent syngeneic renal transplantation or unilateral nephrectomy (UNX) and were deprived of water for 24 hours at six weeks after the operation when histological and functional compensation of the intact kidney was complete. Blood and urinary samples were collected before and after dehydration. The amount of rBSC1 or AQP2 mRNA was measured using competitive polymerase chain reaction (PCR) by inducing a point mutation at the middle of PCR product for rBSC1 or by deleting 180 bp from 780 bp PCR product for AQP2, respectively. The protein expression was examined by Western blot analysis. Results. Both groups of rats demonstrated the same levels of compensatory renal hypertrophy (∼60% weight increase) and plasma creatinine values. Histological examination revealed enlarged glomeruli and tubules, but no findings of ischemic damage, such as tubular atrophy or interstitial changes. Urinary concentration was noted in the UNX rats but not in rats with kidney grafts. Competitive PCR demonstrated that dehydration did not increase rBSC1 and AQP2 transcripts in rats with kidney transplantation. Immunoblot analysis confirmed that the marked increase of both rBSC1 and AQP2 proteins was noted only in the remnant kidney of dehydrated rats.

AB - Background. The underlying mechanisms of renal transplant dysfunction are poorly understood. There is little information on tubular function in kidney grafts. The cDNAs encoding kidney-specific cell surface proteins required for renal reabsorption of sodium (sodium cotransporter in thick ascending limb of Henle, rBSC1) and water (apical water channel in collecting duct, AQP2) have been recently identified. Since transcripts of these proteins are up-regulated in dehydration in association with maximal concentration of urine, we examined urinary concentrating ability and expression levels of mRNA of these proteins in kidney isografts. Methods. Male Sprague-Dawley rats underwent syngeneic renal transplantation or unilateral nephrectomy (UNX) and were deprived of water for 24 hours at six weeks after the operation when histological and functional compensation of the intact kidney was complete. Blood and urinary samples were collected before and after dehydration. The amount of rBSC1 or AQP2 mRNA was measured using competitive polymerase chain reaction (PCR) by inducing a point mutation at the middle of PCR product for rBSC1 or by deleting 180 bp from 780 bp PCR product for AQP2, respectively. The protein expression was examined by Western blot analysis. Results. Both groups of rats demonstrated the same levels of compensatory renal hypertrophy (∼60% weight increase) and plasma creatinine values. Histological examination revealed enlarged glomeruli and tubules, but no findings of ischemic damage, such as tubular atrophy or interstitial changes. Urinary concentration was noted in the UNX rats but not in rats with kidney grafts. Competitive PCR demonstrated that dehydration did not increase rBSC1 and AQP2 transcripts in rats with kidney transplantation. Immunoblot analysis confirmed that the marked increase of both rBSC1 and AQP2 proteins was noted only in the remnant kidney of dehydrated rats.

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U2 - 10.1046/j.1523-1755.2001.060002672.x

DO - 10.1046/j.1523-1755.2001.060002672.x

M3 - Article

VL - 60

SP - 672

EP - 679

JO - Kidney International

JF - Kidney International

SN - 0085-2538

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