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CYP26A1 knockout embryonic stem cells exhibit reduced differentiation and growth arrest in response to retinoic acid
CYP26A1, a cytochrome P450 enzyme, metabolizes all-trans-retinoic acid (RA) into polar metabolites, e.g. 4-oxo-RA and 4-OH-RA. To determine if altering RA metabolism affects embryonic stem (ES) cell differentiation, we disrupted both alleles of Cyp26a1 by homologous recombination. CYP26a1−/− ES cell...
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Published in: | Developmental biology 2008-03, Vol.315 (2), p.331-354 |
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description | CYP26A1, a cytochrome P450 enzyme, metabolizes all-trans-retinoic acid (RA) into polar metabolites, e.g. 4-oxo-RA and 4-OH-RA. To determine if altering RA metabolism affects embryonic stem (ES) cell differentiation, we disrupted both alleles of Cyp26a1 by homologous recombination. CYP26a1−/− ES cells had a 11.0±3.2-fold higher intracellular RA concentration than Wt ES cells after RA treatment for 48 h. RA-treated CYP26A1−/− ES cells exhibited 2–3 fold higher mRNA levels of Hoxa1, a primary RA target gene, than Wt ES cells. Despite increased intracellular RA levels, CYP26a1−/− ES cells were more resistant than Wt ES cells to RA-induced proliferation arrest. Transcripts for parietal endodermal differentiation markers, including laminin, J6(Hsp 47), and J31(SPARC, osteonectin) were expressed at lower levels in RA-treated CYP26a1−/− ES cells, indicating that the lack of CYP26A1 activity inhibits RA-associated differentiation. Microarray analyses revealed that RA-treated CYP26A1−/− ES cells exhibited lower mRNA levels than Wt ES cells for genes involved in differentiation, particularly in neural (Epha4, Pmp22, Nrp1, Gap43, Ndn) and smooth muscle differentiation (Madh3, Nrp1, Tagln Calponin, Caldesmon1). In contrast, genes involved in the stress response (e.g. Tlr2, Stk2, Fcgr2b, Bnip3, Pdk1) were expressed at higher levels in CYP26A1−/− than in Wt ES cells without RA. Collectively, our results show that CYP26A1 activity regulates intracellular RA levels, cell proliferation, transcriptional regulation of primary RA target genes, and ES cell differentiation to parietal endoderm. |
doi_str_mv | 10.1016/j.ydbio.2007.12.021 |
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To determine if altering RA metabolism affects embryonic stem (ES) cell differentiation, we disrupted both alleles of Cyp26a1 by homologous recombination. CYP26a1−/− ES cells had a 11.0±3.2-fold higher intracellular RA concentration than Wt ES cells after RA treatment for 48 h. RA-treated CYP26A1−/− ES cells exhibited 2–3 fold higher mRNA levels of Hoxa1, a primary RA target gene, than Wt ES cells. Despite increased intracellular RA levels, CYP26a1−/− ES cells were more resistant than Wt ES cells to RA-induced proliferation arrest. Transcripts for parietal endodermal differentiation markers, including laminin, J6(Hsp 47), and J31(SPARC, osteonectin) were expressed at lower levels in RA-treated CYP26a1−/− ES cells, indicating that the lack of CYP26A1 activity inhibits RA-associated differentiation. Microarray analyses revealed that RA-treated CYP26A1−/− ES cells exhibited lower mRNA levels than Wt ES cells for genes involved in differentiation, particularly in neural (Epha4, Pmp22, Nrp1, Gap43, Ndn) and smooth muscle differentiation (Madh3, Nrp1, Tagln Calponin, Caldesmon1). In contrast, genes involved in the stress response (e.g. Tlr2, Stk2, Fcgr2b, Bnip3, Pdk1) were expressed at higher levels in CYP26A1−/− than in Wt ES cells without RA. Collectively, our results show that CYP26A1 activity regulates intracellular RA levels, cell proliferation, transcriptional regulation of primary RA target genes, and ES cell differentiation to parietal endoderm.</description><identifier>ISSN: 0012-1606</identifier><identifier>EISSN: 1095-564X</identifier><identifier>DOI: 10.1016/j.ydbio.2007.12.021</identifier><identifier>PMID: 18241852</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Base Sequence ; Cell Differentiation - drug effects ; Cell Line ; Cell Proliferation - drug effects ; CYP26A1 ; Cytochrome P-450 Enzyme System - deficiency ; Cytochrome P-450 Enzyme System - genetics ; Cytochrome P-450 Enzyme System - metabolism ; Cytochrome P450 ; DNA Primers - genetics ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - drug effects ; Embryonic Stem Cells - enzymology ; Endoderm differentiation ; Gene Expression - drug effects ; Gene Targeting ; Homeodomain Proteins - genetics ; Knockout ; Laminin - genetics ; Leukemia Inhibitory Factor - pharmacology ; Metabolism ; Mice ; Mice, Knockout ; Microarray ; Models, Biological ; Myocytes, Smooth Muscle - cytology ; Myocytes, Smooth Muscle - drug effects ; Myocytes, Smooth Muscle - metabolism ; Neurons - cytology ; Neurons - drug effects ; Neurons - metabolism ; Oligonucleotide Array Sequence Analysis ; Retinoic acid ; Retinoic Acid 4-Hydroxylase ; Review ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Stem cells ; Transcription Factors - genetics ; Tretinoin - metabolism ; Tretinoin - pharmacology ; Vitamin A</subject><ispartof>Developmental biology, 2008-03, Vol.315 (2), p.331-354</ispartof><rights>2007 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c464t-d8cf7b0ed36a66afecb153bf6b62535b82191e24452ccba78d677cd9c7bed1883</citedby><cites>FETCH-LOGICAL-c464t-d8cf7b0ed36a66afecb153bf6b62535b82191e24452ccba78d677cd9c7bed1883</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18241852$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Langton, Simne</creatorcontrib><creatorcontrib>Gudas, Lorraine J.</creatorcontrib><title>CYP26A1 knockout embryonic stem cells exhibit reduced differentiation and growth arrest in response to retinoic acid</title><title>Developmental biology</title><addtitle>Dev Biol</addtitle><description>CYP26A1, a cytochrome P450 enzyme, metabolizes all-trans-retinoic acid (RA) into polar metabolites, e.g. 4-oxo-RA and 4-OH-RA. To determine if altering RA metabolism affects embryonic stem (ES) cell differentiation, we disrupted both alleles of Cyp26a1 by homologous recombination. CYP26a1−/− ES cells had a 11.0±3.2-fold higher intracellular RA concentration than Wt ES cells after RA treatment for 48 h. RA-treated CYP26A1−/− ES cells exhibited 2–3 fold higher mRNA levels of Hoxa1, a primary RA target gene, than Wt ES cells. Despite increased intracellular RA levels, CYP26a1−/− ES cells were more resistant than Wt ES cells to RA-induced proliferation arrest. Transcripts for parietal endodermal differentiation markers, including laminin, J6(Hsp 47), and J31(SPARC, osteonectin) were expressed at lower levels in RA-treated CYP26a1−/− ES cells, indicating that the lack of CYP26A1 activity inhibits RA-associated differentiation. Microarray analyses revealed that RA-treated CYP26A1−/− ES cells exhibited lower mRNA levels than Wt ES cells for genes involved in differentiation, particularly in neural (Epha4, Pmp22, Nrp1, Gap43, Ndn) and smooth muscle differentiation (Madh3, Nrp1, Tagln Calponin, Caldesmon1). In contrast, genes involved in the stress response (e.g. Tlr2, Stk2, Fcgr2b, Bnip3, Pdk1) were expressed at higher levels in CYP26A1−/− than in Wt ES cells without RA. Collectively, our results show that CYP26A1 activity regulates intracellular RA levels, cell proliferation, transcriptional regulation of primary RA target genes, and ES cell differentiation to parietal endoderm.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Line</subject><subject>Cell Proliferation - drug effects</subject><subject>CYP26A1</subject><subject>Cytochrome P-450 Enzyme System - deficiency</subject><subject>Cytochrome P-450 Enzyme System - genetics</subject><subject>Cytochrome P-450 Enzyme System - metabolism</subject><subject>Cytochrome P450</subject><subject>DNA Primers - genetics</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - drug effects</subject><subject>Embryonic Stem Cells - enzymology</subject><subject>Endoderm differentiation</subject><subject>Gene Expression - drug effects</subject><subject>Gene Targeting</subject><subject>Homeodomain Proteins - genetics</subject><subject>Knockout</subject><subject>Laminin - genetics</subject><subject>Leukemia Inhibitory Factor - pharmacology</subject><subject>Metabolism</subject><subject>Mice</subject><subject>Mice, Knockout</subject><subject>Microarray</subject><subject>Models, Biological</subject><subject>Myocytes, Smooth Muscle - cytology</subject><subject>Myocytes, Smooth Muscle - drug effects</subject><subject>Myocytes, Smooth Muscle - metabolism</subject><subject>Neurons - cytology</subject><subject>Neurons - drug effects</subject><subject>Neurons - metabolism</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Retinoic acid</subject><subject>Retinoic Acid 4-Hydroxylase</subject><subject>Review</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Stem cells</subject><subject>Transcription Factors - genetics</subject><subject>Tretinoin - metabolism</subject><subject>Tretinoin - pharmacology</subject><subject>Vitamin A</subject><issn>0012-1606</issn><issn>1095-564X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><recordid>eNqFkU1vEzEQhi0EomnhFyAhn7jt4vHu2t4DhyriS6oEB5DgZPljljpN1sH2FvLvcUgkbvTkOTzzjmceQl4Aa4GBeL1pD96G2HLGZAu8ZRwekRWwcWgG0X97TFaMAW9AMHFBLnPeMMY6pbqn5AIU70ENfEXK-vtnLq6B3s3R3cWlUNzZdIhzcDQX3FGH222m-Ps22FBoQr849NSHacKEcwmmhDhTM3v6I8Vf5ZaalDAXGuYK532cM9ISa13CHGuoccE_I08ms834_Pxeka_v3n5Zf2huPr3_uL6-aVwv-tJ45SZpGfpOGCHMhM7C0NlJWMGHbrCKwwjI-37gzlkjlRdSOj86adFD3fSKvDrl7lP8udRf6V3Ix4XMjHHJWrJOSqX4gyAHNsgRugdBGCUoMbAKdifQpZhzwknvU9iZdNDA9FGf3ui_-vRRnwauq77a9fIcv9gd-n89Z18VeHMCsJ7tPmDS2QWcq5KQ0BXtY_jvgD8GjK5V</recordid><startdate>20080315</startdate><enddate>20080315</enddate><creator>Langton, Simne</creator><creator>Gudas, Lorraine J.</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20080315</creationdate><title>CYP26A1 knockout embryonic stem cells exhibit reduced differentiation and growth arrest in response to retinoic acid</title><author>Langton, Simne ; Gudas, Lorraine J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c464t-d8cf7b0ed36a66afecb153bf6b62535b82191e24452ccba78d677cd9c7bed1883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Line</topic><topic>Cell Proliferation - drug effects</topic><topic>CYP26A1</topic><topic>Cytochrome P-450 Enzyme System - deficiency</topic><topic>Cytochrome P-450 Enzyme System - genetics</topic><topic>Cytochrome P-450 Enzyme System - metabolism</topic><topic>Cytochrome P450</topic><topic>DNA Primers - genetics</topic><topic>Embryonic Stem Cells - cytology</topic><topic>Embryonic Stem Cells - drug effects</topic><topic>Embryonic Stem Cells - enzymology</topic><topic>Endoderm differentiation</topic><topic>Gene Expression - drug effects</topic><topic>Gene Targeting</topic><topic>Homeodomain Proteins - genetics</topic><topic>Knockout</topic><topic>Laminin - genetics</topic><topic>Leukemia Inhibitory Factor - pharmacology</topic><topic>Metabolism</topic><topic>Mice</topic><topic>Mice, Knockout</topic><topic>Microarray</topic><topic>Models, Biological</topic><topic>Myocytes, Smooth Muscle - cytology</topic><topic>Myocytes, Smooth Muscle - drug effects</topic><topic>Myocytes, Smooth Muscle - metabolism</topic><topic>Neurons - cytology</topic><topic>Neurons - drug effects</topic><topic>Neurons - metabolism</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>Retinoic acid</topic><topic>Retinoic Acid 4-Hydroxylase</topic><topic>Review</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Stem cells</topic><topic>Transcription Factors - genetics</topic><topic>Tretinoin - metabolism</topic><topic>Tretinoin - pharmacology</topic><topic>Vitamin A</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Langton, Simne</creatorcontrib><creatorcontrib>Gudas, Lorraine J.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Developmental biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Langton, Simne</au><au>Gudas, Lorraine J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>CYP26A1 knockout embryonic stem cells exhibit reduced differentiation and growth arrest in response to retinoic acid</atitle><jtitle>Developmental biology</jtitle><addtitle>Dev Biol</addtitle><date>2008-03-15</date><risdate>2008</risdate><volume>315</volume><issue>2</issue><spage>331</spage><epage>354</epage><pages>331-354</pages><issn>0012-1606</issn><eissn>1095-564X</eissn><abstract>CYP26A1, a cytochrome P450 enzyme, metabolizes all-trans-retinoic acid (RA) into polar metabolites, e.g. 4-oxo-RA and 4-OH-RA. To determine if altering RA metabolism affects embryonic stem (ES) cell differentiation, we disrupted both alleles of Cyp26a1 by homologous recombination. CYP26a1−/− ES cells had a 11.0±3.2-fold higher intracellular RA concentration than Wt ES cells after RA treatment for 48 h. RA-treated CYP26A1−/− ES cells exhibited 2–3 fold higher mRNA levels of Hoxa1, a primary RA target gene, than Wt ES cells. Despite increased intracellular RA levels, CYP26a1−/− ES cells were more resistant than Wt ES cells to RA-induced proliferation arrest. Transcripts for parietal endodermal differentiation markers, including laminin, J6(Hsp 47), and J31(SPARC, osteonectin) were expressed at lower levels in RA-treated CYP26a1−/− ES cells, indicating that the lack of CYP26A1 activity inhibits RA-associated differentiation. Microarray analyses revealed that RA-treated CYP26A1−/− ES cells exhibited lower mRNA levels than Wt ES cells for genes involved in differentiation, particularly in neural (Epha4, Pmp22, Nrp1, Gap43, Ndn) and smooth muscle differentiation (Madh3, Nrp1, Tagln Calponin, Caldesmon1). In contrast, genes involved in the stress response (e.g. Tlr2, Stk2, Fcgr2b, Bnip3, Pdk1) were expressed at higher levels in CYP26A1−/− than in Wt ES cells without RA. Collectively, our results show that CYP26A1 activity regulates intracellular RA levels, cell proliferation, transcriptional regulation of primary RA target genes, and ES cell differentiation to parietal endoderm.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>18241852</pmid><doi>10.1016/j.ydbio.2007.12.021</doi><tpages>24</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Base Sequence Cell Differentiation - drug effects Cell Line Cell Proliferation - drug effects CYP26A1 Cytochrome P-450 Enzyme System - deficiency Cytochrome P-450 Enzyme System - genetics Cytochrome P-450 Enzyme System - metabolism Cytochrome P450 DNA Primers - genetics Embryonic Stem Cells - cytology Embryonic Stem Cells - drug effects Embryonic Stem Cells - enzymology Endoderm differentiation Gene Expression - drug effects Gene Targeting Homeodomain Proteins - genetics Knockout Laminin - genetics Leukemia Inhibitory Factor - pharmacology Metabolism Mice Mice, Knockout Microarray Models, Biological Myocytes, Smooth Muscle - cytology Myocytes, Smooth Muscle - drug effects Myocytes, Smooth Muscle - metabolism Neurons - cytology Neurons - drug effects Neurons - metabolism Oligonucleotide Array Sequence Analysis Retinoic acid Retinoic Acid 4-Hydroxylase Review RNA, Messenger - genetics RNA, Messenger - metabolism Stem cells Transcription Factors - genetics Tretinoin - metabolism Tretinoin - pharmacology Vitamin A |
title | CYP26A1 knockout embryonic stem cells exhibit reduced differentiation and growth arrest in response to retinoic acid |
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