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Altered sodium pump alpha and gamma subunit gene expression in nephron segments from hypertensive rats

 

作者: Amanda Hayward,   Cruz Hinojos,   Barbara Nurowska,   Aveline Hewetson,   Sandra Sabatini,   Peter Oefner,   Peter Doris,  

 

期刊: Journal of Hypertension  (OVID Available online 1999)
卷期: Volume 17, issue 8  

页码: 1081-1087

 

ISSN:0263-6352

 

年代: 1999

 

出版商: OVID

 

关键词: hypertension;development;renal sodium reabsorption;quantitative RT-PCR;proximal tubule;‘sodium potassium-ATPase’

 

数据来源: OVID

 

摘要:

ObjectiveTo determine the qualitative and quantitative expression of alpha and gamma sodium pump subunits in whole kidney and nephron segment RNA from Sprague Dawley rats, spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats.DesignA novel reverse transcription polymerase chain reaction technique was devised which provides accurate and precise measurement of the number of molecules of specific transcript abundance, a measurement of gene expression. This allows the quantitative comparison of multiple samples across multiple subjects and, since the estimates are accurate rather than relative, can also be used to make quantitative comparisons across expressed genes, such as isoforms and subunits of the heterotrimeric renal sodium pump.MethodsWe examined which catalytic isoforms were expressed and then quantified transcript abundance in whole kidney and convoluted and straight segments of the proximal tubule.ResultsAlpha 1 and gamma transcripts, but not alpha 2, alpha 3 or alpha 4 isoforms, were consistently observed in nephron segments. Levels of alpha 1 were lower in kidney RNA from 15–16-week-old SHR than in WKY rats of the same age (P= 0.001), but were not different between SHR and WKY in 4–5-week-old animals. No significant difference was observed in gamma subunit abundance in kidney RNA from 4±5-week-old animals; however, at 15–16 weeks, the expression in SHR was one-third that in WKY rats (P= 0.003). In proximal convoluted tubules from 4–5-week-old animals, the level of alpha 1 RNA expression was lower (P= 0.03) in SHR than in WKY rats. In addition, levels of alpha 1 in proximal straight tubule from the 4–5-week-old SHR were also lower than in WKY rats (P= 0.02). This difference was even greater in 15–16-week-old animals: in SHR, alpha 1 expression was less than 20% of the level of expression in WKY rats (P= 0.0003). Expression of the gamma subunit exhibited a similar pattern of downregulation in SHR. In RNA from proximal convoluted tubules and proximal straight tubules from both 4–5- and 15–16-week-old animals, expression of the gamma subunit was demonstrated to be significantly lower in SHR than in WKY rats.ConclusionThe results indicate a coordinate reduction in the abundance of sodium pump alpha and gamma subunits in the proximal tubules of SHR, which occurs early during the development of hypertension.

 

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