The expression of renin-binding protein and renin in the kidneys of rats with two-kidney one-clip hypertension.
OBJECTIVE
To test the hypothesis that renin-binding protein (RnBP) is involved in modulating the intracellular processing or release of renin, we examined the expression of RnBP in clipped and contralateral kidneys of rats with two-kidney one-clip hypertension, and in left and right kidneys from sham-operated control rats.


METHODS
Kidneys from rats with two-kidney one-clip hypertension and from control rats were either snap-frozen for extraction of mRNA or fixed for in-situ hybridization and immunochemistry. Reverse-transcription polymerase chain reaction on renal mRNA was performed using primers for renin, RnBP, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and angiotensin-converting enzyme (ACE). In addition, renal total RNA was analysed by Northern blotting for RnBP, GAPDH and angiotensin II type 1A (AT1A) receptor mRNA, and the intensity of the bands was measured by laser densitometry. In situ hybridization for renin mRNA was carried out using digoxygenin-labelled antisense oligonucleotides and for RnBP using labelled antisense oligonucleotides and an antisense riboprobe. Controls included sections treated with RNase and sections stained with sense oligonucleotides.


RESULTS
The level of expression of mRNA for RnBP is similar in clipped and contralateral kidneys of renal hypertensive rats; in contrast, renin mRNA expression is upregulated in the clipped kidney. Renin-binding protein is expressed mainly in renal tubules and collecting ducts unlike renin, which is expressed in the glomerular afferent arteriole. We did not detect lateralization of expression for ACE or the AT1A receptor between clipped and contralateral kidneys.


CONCLUSIONS
Renin-binding protein expression is unchanged between clipped and contralateral kidneys. Therefore, a physiological stimulus that upregulates renin gene expression in clipped kidneys does not affect RnBP expression. The main sites of RnBP expression are the renal tubules and collecting ducts; in contast renin is expressed at the glomerular pole. The results show that RnBP is not colocalized or coregulated with renin in this model of hypertension.
Leckie
B J
BJ
Department of Medicine and Therapeutics, Leicester Royal Infirmary, UK. bjl2@le.ac.uk
Lacy
P S
PS
Lidder
S
S
D000818:Animals; D015152:Blotting, Northern; D002238:Carbohydrate Epimerases / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D006977:Hypertension, Renal / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D017403:In Situ Hybridization; D007668:Kidney / Q000378:metabolism* / Q000473:pathology; D008026:Ligation; D007703:Peptidyl-Dipeptidase A / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D011921:Rats, Inbred WKY; D044140:Receptor, Angiotensin, Type 1; D011945:Receptors, Angiotensin / Q000235:genetics / Q000378:metabolism; D012077:Renal Artery / Q000601:surgery; D012083:Renin / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002352:Carrier Proteins; D017931:DNA Primers; D012333:RNA, Messenger; D044140:Receptor, Angiotensin, Type 1; D011945:Receptors, Angiotensin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007703:Peptidyl-Dipeptidase A; D012083:Renin; D002238:Carbohydrate Epimerases; C024519:N-acyl-D-glucosamine 2-epimerase; C498258:Renbp protein, rat