Cytochrome P450-2C11 mRNA is not expressed in endothelial cells dissected from rat renal arterioles.
BACKGROUND
Cytochrome P450 (CYP) isoenzymes (CYP2C and CYP2J) are involved in the production of epoxyeicosatrienoic acids, which are postulated as endothelium-derived hyperpolarizing factors (EDHFs). We hypothesized that if CYP2C11 is involved in the EDHF-mediated responses, its mRNA should be expressed in endothelial cells. We, therefore, examined the mRNA expression of CYP2C11 in endothelial cells of renal arterioles.


METHODS
Laser microdissection was applied to isolate endothelial cells from the renal arterioles of 4 male and 4 female Wistar rats. As a positive control of CYP2C11 expression, hepatocytes were also dissected from these rats. RNA was isolated and real-time quantitative polymerase chain reaction (Q-PCR) analysis was applied.


RESULTS
Q-PCR analysis showed that CYP2C11 mRNA was not expressed in laser microdissected endothelial cells of renal arterioles of male and female rats. CYP2C11 mRNA expression was highly abundant in hepatocytes dissected from male livers, but in female livers hardly any CYP2C11 mRNA was detected.


CONCLUSIONS
We have shown that endothelial cells can be dissected from small renal arterioles by laser microdissection to study the mRNA expression of specific genes by Q-PCR. Using this novel tool, we demonstrated that the CYP2C11 mRNA was not expressed in the endothelial cells of renal arterioles. Therefore, we speculate that CYP2C11 does not contribute to the EDHF-mediated responses in renal arterioles.
Heil
Sandra G
SG
Laboratory of Pediatrics and Neurology, University Medical Center Nijmegen, Nijmegen, The Netherlands.
De Vriese
An S
AS
Kluijtmans
Leo A J
LA
Dijkman
Henry
H
van Strien
Denise
D
Akkers
Robert
R
Blom
Henk J
HJ
D000818:Animals; D001160:Arterioles / Q000201:enzymology*; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D000072467:Cytochrome P450 Family 2; D042783:Endothelial Cells / Q000201:enzymology*; D005260:Female; D066298:In Vitro Techniques; D007668:Kidney / Q000098:blood supply* / Q000201:enzymology*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D039181:Steroid 16-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*
D012333:RNA, Messenger; D001189:Aryl Hydrocarbon Hydroxylases; C450279:CYP2C11 protein, rat; D000072467:Cytochrome P450 Family 2; D039181:Steroid 16-alpha-Hydroxylase