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The prothrombin gene is expressed in the rat kidney: Implications for urolithiasis research

Author:
Grover, Phulwinder K., Dogra, Satish C., Davidson, Benjamin P., Stapleton, Alan M. F., Ryall, Rosemary L.
Source:
European journal of biochemistry 2000 v.267 no.1 pp. 61-67
ISSN:
0014-2956
Subject:
agarose, animal models, biosynthesis, calcium, calcium oxalate, crystals, gel electrophoresis, gene expression, genes, glyceraldehyde-3-phosphate dehydrogenase, humans, kidneys, liver, messenger RNA, pathogenesis, prothrombin, rats, reverse transcriptase polymerase chain reaction, sequence analysis, thrombin, urinary calculi, urine
Abstract:
There is considerable interest in determining the role of prothrombin fragments, especially urinary prothrombin fragment 1 (UPTF1), in the pathogenesis of calcium oxalate (CaOx) urinary calculi. This fragment is present in abundance in the matrix of CaOx crystals generated in human urine in vitro and has also been detected in human urinary stones containing calcium. More recently, prothrombin gene expression has been reported in the human kidney. However, studies examining the renal biosynthesis of prothrombin or perhaps only its fragments during experimental lithogenesis, and in consequence, the role of UPTF1 in stone formation, cannot be carried out in humans. The aim of this investigation therefore was to determine whether prothrombin gene expression is present in the rat kidney. Total RNA was isolated from the kidneys and livers of 12 rats. Using reverse transcriptase PCR, mRNAs corresponding to the thrombin and fragment 1 + 2 (F1+2) regions of prothrombin were analysed by agarose gel electrophoresis. The expression of glyceraldehyde 3‐phosphate dehydrogenase was also examined to determine whether the quality of the tissue mRNAs was adequate for analyses. The amplified products were identified by sequence analysis. All kidneys displayed evidence of expression of the thrombin and F1+2 domains of the prothrombin gene. Furthermore, the sequences of these PCR‐derived products from kidney were identical to those from liver. This suggests that the prothrombins secreted by these two organs are identical. The fact that prothrombin biosynthesis occurs in both the human and rat kidney presents an opportunity for using established rat models of stone disease to evaluate the influence of lithogenic conditions on prothrombin gene expression, and the potential role of UPTF1 in vivo.
Agid:
96055