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 expr...
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| Veröffentlicht in: | Nephron 2005-02, Vol.99 (2), p.p43-p49 |
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| creator | Heil, Sandra G. De Vriese, An S. Kluijtmans, Leo A.J. Dijkman, Henry van Strien, Denise Akkers, Robert Blom, Henk J. |
| description | 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. Conclusion: 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. |
| doi_str_mv | 10.1159/000083135 |
| format | Article |
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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. Conclusion: 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.</description><identifier>ISSN: 1660-2137</identifier><identifier>ISSN: 1660-8151</identifier><identifier>ISSN: 0028-2766</identifier><identifier>EISSN: 1660-2137</identifier><identifier>EISSN: 2235-3186</identifier><identifier>DOI: 10.1159/000083135</identifier><identifier>PMID: 15637425</identifier><identifier>CODEN: NPRNAY</identifier><language>eng</language><publisher>Basel, Switzerland: S. Karger AG</publisher><subject>Animals ; Arterioles - enzymology ; Aryl Hydrocarbon Hydroxylases - genetics ; Aryl Hydrocarbon Hydroxylases - metabolism ; Cells, Cultured ; Cytochrome P450 Family 2 ; Endothelial Cells - enzymology ; Female ; In Vitro Techniques ; Kidney - blood supply ; Kidney - enzymology ; Male ; Original Paper ; Rats ; Rats, Wistar ; RNA, Messenger - metabolism ; Steroid 16-alpha-Hydroxylase - genetics ; Steroid 16-alpha-Hydroxylase - metabolism</subject><ispartof>Nephron, 2005-02, Vol.99 (2), p.p43-p49</ispartof><rights>2005 S. Karger AG, Basel</rights><rights>Copyright (c) 2005 S. Karger AG, Basel.</rights><rights>Copyright (c) 2005 S. Karger AG, Basel</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c276t-a46cf06acc51fbdbe16ed66d75fa6b8c03d6c39034f3570ff3793d1f734dc0a73</citedby><cites>FETCH-LOGICAL-c276t-a46cf06acc51fbdbe16ed66d75fa6b8c03d6c39034f3570ff3793d1f734dc0a73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,2427,27922,27923</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15637425$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Heil, Sandra G.</creatorcontrib><creatorcontrib>De Vriese, An S.</creatorcontrib><creatorcontrib>Kluijtmans, Leo A.J.</creatorcontrib><creatorcontrib>Dijkman, Henry</creatorcontrib><creatorcontrib>van Strien, Denise</creatorcontrib><creatorcontrib>Akkers, Robert</creatorcontrib><creatorcontrib>Blom, Henk J.</creatorcontrib><title>Cytochrome P450-2C11 mRNA Is Not Expressed in Endothelial Cells Dissected from Rat Renal Arterioles</title><title>Nephron</title><addtitle>Nephron Physiol</addtitle><description>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. Conclusion: 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.</description><subject>Animals</subject><subject>Arterioles - enzymology</subject><subject>Aryl Hydrocarbon Hydroxylases - genetics</subject><subject>Aryl Hydrocarbon Hydroxylases - metabolism</subject><subject>Cells, Cultured</subject><subject>Cytochrome P450 Family 2</subject><subject>Endothelial Cells - enzymology</subject><subject>Female</subject><subject>In Vitro Techniques</subject><subject>Kidney - blood supply</subject><subject>Kidney - enzymology</subject><subject>Male</subject><subject>Original Paper</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>RNA, Messenger - metabolism</subject><subject>Steroid 16-alpha-Hydroxylase - genetics</subject><subject>Steroid 16-alpha-Hydroxylase - metabolism</subject><issn>1660-2137</issn><issn>1660-8151</issn><issn>0028-2766</issn><issn>1660-2137</issn><issn>2235-3186</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNqF0ctKAzEUBuAgiq3VhWtBggvBRTX3tMsyVi1IlaLrIZOLnTozqckU7NsbaanixmwSOF9-TnIAOMXoGmM-vEFpDSimfA90sRCoTzCV-7_OHXAU4wIhQhjDh6CDuaCSEd4FOlu3Xs-Dry18ZjzxDGNYz6YjOIlw6ls4_lwGG6M1sGzguDG-nduqVBXMbFVFeFummm5T2aUQOFMtnNkmlUehtaH0lY3H4MCpKtqT7d4Dr3fjl-yh__h0P8lGj31NpGj7igntkFBac-wKU1gsrBHCSO6UKAYaUSM0HSLKHOUSOUflkBrsJGVGIyVpD1xucpfBf6xsbPO6jDp1qRrrVzEXkg6QTP_xHyQYM8IET_DiD1z4VUivS0YyRsRgSBK62iAdfIzBunwZylqFdY5R_j2ffDefZM-3gauituZHbgeSwNkGvKvwZsMObK5_Aa4fkJs</recordid><startdate>20050201</startdate><enddate>20050201</enddate><creator>Heil, Sandra G.</creator><creator>De Vriese, An S.</creator><creator>Kluijtmans, Leo A.J.</creator><creator>Dijkman, Henry</creator><creator>van Strien, Denise</creator><creator>Akkers, Robert</creator><creator>Blom, Henk J.</creator><general>S. 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enzymology</topic><topic>Aryl Hydrocarbon Hydroxylases - genetics</topic><topic>Aryl Hydrocarbon Hydroxylases - metabolism</topic><topic>Cells, Cultured</topic><topic>Cytochrome P450 Family 2</topic><topic>Endothelial Cells - enzymology</topic><topic>Female</topic><topic>In Vitro Techniques</topic><topic>Kidney - blood supply</topic><topic>Kidney - enzymology</topic><topic>Male</topic><topic>Original Paper</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>RNA, Messenger - metabolism</topic><topic>Steroid 16-alpha-Hydroxylase - genetics</topic><topic>Steroid 16-alpha-Hydroxylase - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Heil, Sandra G.</creatorcontrib><creatorcontrib>De Vriese, An S.</creatorcontrib><creatorcontrib>Kluijtmans, Leo A.J.</creatorcontrib><creatorcontrib>Dijkman, Henry</creatorcontrib><creatorcontrib>van Strien, Denise</creatorcontrib><creatorcontrib>Akkers, Robert</creatorcontrib><creatorcontrib>Blom, Henk J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Immunology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>SIRS Editorial</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Nephron</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Heil, Sandra G.</au><au>De Vriese, An S.</au><au>Kluijtmans, Leo A.J.</au><au>Dijkman, Henry</au><au>van Strien, Denise</au><au>Akkers, Robert</au><au>Blom, Henk J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cytochrome P450-2C11 mRNA Is Not Expressed in Endothelial Cells Dissected from Rat Renal Arterioles</atitle><jtitle>Nephron</jtitle><addtitle>Nephron Physiol</addtitle><date>2005-02-01</date><risdate>2005</risdate><volume>99</volume><issue>2</issue><spage>p43</spage><epage>p49</epage><pages>p43-p49</pages><issn>1660-2137</issn><issn>1660-8151</issn><issn>0028-2766</issn><eissn>1660-2137</eissn><eissn>2235-3186</eissn><coden>NPRNAY</coden><abstract>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. Conclusion: 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.</abstract><cop>Basel, Switzerland</cop><pub>S. Karger AG</pub><pmid>15637425</pmid><doi>10.1159/000083135</doi><tpages>1</tpages></addata></record> |
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| subjects | Animals Arterioles - enzymology Aryl Hydrocarbon Hydroxylases - genetics Aryl Hydrocarbon Hydroxylases - metabolism Cells, Cultured Cytochrome P450 Family 2 Endothelial Cells - enzymology Female In Vitro Techniques Kidney - blood supply Kidney - enzymology Male Original Paper Rats Rats, Wistar RNA, Messenger - metabolism Steroid 16-alpha-Hydroxylase - genetics Steroid 16-alpha-Hydroxylase - metabolism |
| title | Cytochrome P450-2C11 mRNA Is Not Expressed in Endothelial Cells Dissected from Rat Renal Arterioles |
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