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Expression and regulation of stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) in equine and bovine preovulatory follicles
•hCG induces CXCR4 expression in equine and bovine preovulatory follicles.•The hCG-dependent induction of CXCR4 occurs in both granulosa and theca cells.•Forskolin induces CXCR4 expression in bovine granulosa cell cultures.•Numerous signaling pathways are involved in follicular CXCR4 expression.•SDF...
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| Published in: | Molecular and cellular endocrinology 2014-06, Vol.391 (1-2), p.10-21 |
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| cites | cdi_FETCH-LOGICAL-c419t-6eb08628a1203233375b8b6980781b6f8a6ba66681bb0e7063cfa232dc05bc483 |
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| container_title | Molecular and cellular endocrinology |
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| creator | Sayasith, Khampoun Sirois, Jean |
| description | •hCG induces CXCR4 expression in equine and bovine preovulatory follicles.•The hCG-dependent induction of CXCR4 occurs in both granulosa and theca cells.•Forskolin induces CXCR4 expression in bovine granulosa cell cultures.•Numerous signaling pathways are involved in follicular CXCR4 expression.•SDF1/CXCR4 activation increases the expression of ovulation-regulated genes.
The interaction between stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) has been implicated in leukocyte attraction, tissue remodeling and angiogenesis. The objective of the present study was to characterize the expression and regulation of SDF1 and CXCR4 in equine follicles during the ovulatory process. Equine preovulatory follicles were isolated during estrus 0–39h after hCG treatment. Follicle wall preparations (theca interna with attached granulosa cells) and isolated preparations of granulosa cells and theca interna were obtained, and total RNA extracts were analyzed by RT-PCR/Southern blot. Results showed that levels of CXCR4 transcripts were induced by hCG in follicles at 36h post-hCG (P |
| doi_str_mv | 10.1016/j.mce.2014.04.009 |
| format | article |
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The interaction between stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) has been implicated in leukocyte attraction, tissue remodeling and angiogenesis. The objective of the present study was to characterize the expression and regulation of SDF1 and CXCR4 in equine follicles during the ovulatory process. Equine preovulatory follicles were isolated during estrus 0–39h after hCG treatment. Follicle wall preparations (theca interna with attached granulosa cells) and isolated preparations of granulosa cells and theca interna were obtained, and total RNA extracts were analyzed by RT-PCR/Southern blot. Results showed that levels of CXCR4 transcripts were induced by hCG in follicles at 36h post-hCG (P<0.05 vs 0h), with the induction observed in both granulosa and theca cells. Immunoblotting and immunohistochemical analyses confirmed an increase in CXCR4 protein in follicles after hCG treatment. In contrast, levels of SDF1 transcripts were very low in granulosa cells but high in theca interna cells throughout most of the ovulatory period. Studies in vivo performed with bovine preovulatory follicles collected 0–24h post-hCG revealed a marked and significant up-regulation of CXCR4 transcripts after hCG (P<0.05), as observed in equine follicles. A similar pattern of CXCR4 mRNA up-regulation was observed in cultures of bovine granulosa cells treated with forskolin (P<0.05). This forskolin-dependent induction of CXCR4 mRNA was suppressed by co-treatment with inhibitors of PKA, ERK1/2 and EGFR, and by the progesterone receptor antagonist RU486 (P<0.05), underscoring the contribution of multiple signaling pathways. In complementary studies, treatment of bovine granulosa cells with EGF or the hypoxia mimetic cobalt chloride significantly increased CXCR4 transcript levels, whereas co-treatment with forskolin and a CXCR4 antagonist repressed the expression of several ovulation-related genes. Collectively, this study describes for the first time the gonadotropin-dependent up-regulation of CXCR4 transcript in ovarian follicles of large monoovulatory species, provides some insights into the regulation of CXCR4 gene expression in granulosa cells, and identifies a potential link between follicular SDF1/CXCR4 activation and the regulation of ovulation-related genes.</description><identifier>ISSN: 0303-7207</identifier><identifier>EISSN: 1872-8057</identifier><identifier>DOI: 10.1016/j.mce.2014.04.009</identifier><identifier>PMID: 24784705</identifier><language>eng</language><publisher>Ireland: Elsevier Ireland Ltd</publisher><subject>Activation ; Amino Acid Sequence ; And ovulation-related genes ; Animals ; Cattle ; Cells, Cultured ; Cellular ; Chemokine CXCL12 - genetics ; Chemokine CXCL12 - metabolism ; Chemokine receptor ; Chlorides ; Chorionic Gonadotropin - administration & dosage ; Cobalt - pharmacology ; Colforsin - pharmacology ; Control ; Epidermal Growth Factor - pharmacology ; Estrus - physiology ; Female ; Gene expression ; Gene Expression Regulation ; Genes ; Gonadotropin ; Granulosa Cells - cytology ; Granulosa Cells - drug effects ; Granulosa Cells - metabolism ; Horses ; Humans ; Inhibitors ; Mifepristone - pharmacology ; Molecular Sequence Data ; Ovulation - physiology ; Ovulatory process ; Preovulatory follicles ; Primary granulosa cells ; Protein Kinase Inhibitors - pharmacology ; Receptors ; Receptors, CXCR4 - agonists ; Receptors, CXCR4 - antagonists & inhibitors ; Receptors, CXCR4 - genetics ; Receptors, CXCR4 - metabolism ; RNA, Messenger - agonists ; RNA, Messenger - antagonists & inhibitors ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Signal Transduction ; Theca Cells - cytology ; Theca Cells - drug effects ; Theca Cells - metabolism</subject><ispartof>Molecular and cellular endocrinology, 2014-06, Vol.391 (1-2), p.10-21</ispartof><rights>2014 Elsevier Ireland Ltd</rights><rights>Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c419t-6eb08628a1203233375b8b6980781b6f8a6ba66681bb0e7063cfa232dc05bc483</citedby><cites>FETCH-LOGICAL-c419t-6eb08628a1203233375b8b6980781b6f8a6ba66681bb0e7063cfa232dc05bc483</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/24784705$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sayasith, Khampoun</creatorcontrib><creatorcontrib>Sirois, Jean</creatorcontrib><title>Expression and regulation of stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) in equine and bovine preovulatory follicles</title><title>Molecular and cellular endocrinology</title><addtitle>Mol Cell Endocrinol</addtitle><description>•hCG induces CXCR4 expression in equine and bovine preovulatory follicles.•The hCG-dependent induction of CXCR4 occurs in both granulosa and theca cells.•Forskolin induces CXCR4 expression in bovine granulosa cell cultures.•Numerous signaling pathways are involved in follicular CXCR4 expression.•SDF1/CXCR4 activation increases the expression of ovulation-regulated genes.
The interaction between stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) has been implicated in leukocyte attraction, tissue remodeling and angiogenesis. The objective of the present study was to characterize the expression and regulation of SDF1 and CXCR4 in equine follicles during the ovulatory process. Equine preovulatory follicles were isolated during estrus 0–39h after hCG treatment. Follicle wall preparations (theca interna with attached granulosa cells) and isolated preparations of granulosa cells and theca interna were obtained, and total RNA extracts were analyzed by RT-PCR/Southern blot. Results showed that levels of CXCR4 transcripts were induced by hCG in follicles at 36h post-hCG (P<0.05 vs 0h), with the induction observed in both granulosa and theca cells. Immunoblotting and immunohistochemical analyses confirmed an increase in CXCR4 protein in follicles after hCG treatment. In contrast, levels of SDF1 transcripts were very low in granulosa cells but high in theca interna cells throughout most of the ovulatory period. Studies in vivo performed with bovine preovulatory follicles collected 0–24h post-hCG revealed a marked and significant up-regulation of CXCR4 transcripts after hCG (P<0.05), as observed in equine follicles. A similar pattern of CXCR4 mRNA up-regulation was observed in cultures of bovine granulosa cells treated with forskolin (P<0.05). This forskolin-dependent induction of CXCR4 mRNA was suppressed by co-treatment with inhibitors of PKA, ERK1/2 and EGFR, and by the progesterone receptor antagonist RU486 (P<0.05), underscoring the contribution of multiple signaling pathways. In complementary studies, treatment of bovine granulosa cells with EGF or the hypoxia mimetic cobalt chloride significantly increased CXCR4 transcript levels, whereas co-treatment with forskolin and a CXCR4 antagonist repressed the expression of several ovulation-related genes. Collectively, this study describes for the first time the gonadotropin-dependent up-regulation of CXCR4 transcript in ovarian follicles of large monoovulatory species, provides some insights into the regulation of CXCR4 gene expression in granulosa cells, and identifies a potential link between follicular SDF1/CXCR4 activation and the regulation of ovulation-related genes.</description><subject>Activation</subject><subject>Amino Acid Sequence</subject><subject>And ovulation-related genes</subject><subject>Animals</subject><subject>Cattle</subject><subject>Cells, Cultured</subject><subject>Cellular</subject><subject>Chemokine CXCL12 - genetics</subject><subject>Chemokine CXCL12 - metabolism</subject><subject>Chemokine receptor</subject><subject>Chlorides</subject><subject>Chorionic Gonadotropin - administration & dosage</subject><subject>Cobalt - pharmacology</subject><subject>Colforsin - pharmacology</subject><subject>Control</subject><subject>Epidermal Growth Factor - pharmacology</subject><subject>Estrus - physiology</subject><subject>Female</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Genes</subject><subject>Gonadotropin</subject><subject>Granulosa Cells - cytology</subject><subject>Granulosa Cells - drug effects</subject><subject>Granulosa Cells - metabolism</subject><subject>Horses</subject><subject>Humans</subject><subject>Inhibitors</subject><subject>Mifepristone - pharmacology</subject><subject>Molecular Sequence Data</subject><subject>Ovulation - physiology</subject><subject>Ovulatory process</subject><subject>Preovulatory follicles</subject><subject>Primary granulosa cells</subject><subject>Protein Kinase Inhibitors - pharmacology</subject><subject>Receptors</subject><subject>Receptors, CXCR4 - agonists</subject><subject>Receptors, CXCR4 - antagonists & inhibitors</subject><subject>Receptors, CXCR4 - genetics</subject><subject>Receptors, CXCR4 - metabolism</subject><subject>RNA, Messenger - agonists</subject><subject>RNA, Messenger - antagonists & inhibitors</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Signal Transduction</subject><subject>Theca Cells - cytology</subject><subject>Theca Cells - drug effects</subject><subject>Theca Cells - metabolism</subject><issn>0303-7207</issn><issn>1872-8057</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><recordid>eNqNkd2KFDEQhYMo7rj6AN5ILmcveqz8dJLGKxl3VVgQ_AHvQjpdrRm7O7NJz-C-iY9r2lm9VKEgqfCdU1QOIU8ZbBgw9Xy3GT1uODC5gVLQ3CMrZjSvDNT6PlmBAFFpDvqMPMp5BwC65uYhOeNSG6mhXpEfl9_3CXMOcaJu6mjCL4fBzUsbe5rnFEc3UI_DUHWYwhE72js_x1Qxuv7w6opd_JL5rzjGb2FCuv28pWOcQ1-sPO4LSSVdl9f38oKGieLNYcEWURuPy7XMj8dlaEy3tI_DEPyA-TF50Lsh45O785x8urr8uH1TXb97_Xb78rrykjVzpbAFo7hxjIPgQghdt6ZVjQFtWKt641TrlFKlaQE1KOF7xwXvPNStl0ack_XJd5_izQHzbMeQl33dhPGQLVNaN9BoWf8brWsGUnHZ_AfKGyMapnVB2Qn1KeacsLf7FEaXbi0Du8Rsd7bEbJeYLZSCxf7Znf2hHbH7o_idawFenAAsX3cMmGz2ASePXSixzLaL4S_2PwHJTbZ9</recordid><startdate>20140625</startdate><enddate>20140625</enddate><creator>Sayasith, Khampoun</creator><creator>Sirois, Jean</creator><general>Elsevier Ireland Ltd</general><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>7X8</scope><scope>7U5</scope><scope>8FD</scope><scope>L7M</scope></search><sort><creationdate>20140625</creationdate><title>Expression and regulation of stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) in equine and bovine preovulatory follicles</title><author>Sayasith, Khampoun ; Sirois, Jean</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c419t-6eb08628a1203233375b8b6980781b6f8a6ba66681bb0e7063cfa232dc05bc483</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Activation</topic><topic>Amino Acid Sequence</topic><topic>And ovulation-related genes</topic><topic>Animals</topic><topic>Cattle</topic><topic>Cells, Cultured</topic><topic>Cellular</topic><topic>Chemokine CXCL12 - genetics</topic><topic>Chemokine CXCL12 - metabolism</topic><topic>Chemokine receptor</topic><topic>Chlorides</topic><topic>Chorionic Gonadotropin - administration & dosage</topic><topic>Cobalt - pharmacology</topic><topic>Colforsin - pharmacology</topic><topic>Control</topic><topic>Epidermal Growth Factor - pharmacology</topic><topic>Estrus - physiology</topic><topic>Female</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Genes</topic><topic>Gonadotropin</topic><topic>Granulosa Cells - cytology</topic><topic>Granulosa Cells - drug effects</topic><topic>Granulosa Cells - metabolism</topic><topic>Horses</topic><topic>Humans</topic><topic>Inhibitors</topic><topic>Mifepristone - pharmacology</topic><topic>Molecular Sequence Data</topic><topic>Ovulation - physiology</topic><topic>Ovulatory process</topic><topic>Preovulatory follicles</topic><topic>Primary granulosa cells</topic><topic>Protein Kinase Inhibitors - pharmacology</topic><topic>Receptors</topic><topic>Receptors, CXCR4 - agonists</topic><topic>Receptors, CXCR4 - antagonists & inhibitors</topic><topic>Receptors, CXCR4 - genetics</topic><topic>Receptors, CXCR4 - metabolism</topic><topic>RNA, Messenger - agonists</topic><topic>RNA, Messenger - antagonists & inhibitors</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>Signal Transduction</topic><topic>Theca Cells - cytology</topic><topic>Theca Cells - drug effects</topic><topic>Theca Cells - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sayasith, Khampoun</creatorcontrib><creatorcontrib>Sirois, Jean</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>Molecular and cellular endocrinology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sayasith, Khampoun</au><au>Sirois, Jean</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression and regulation of stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) in equine and bovine preovulatory follicles</atitle><jtitle>Molecular and cellular endocrinology</jtitle><addtitle>Mol Cell Endocrinol</addtitle><date>2014-06-25</date><risdate>2014</risdate><volume>391</volume><issue>1-2</issue><spage>10</spage><epage>21</epage><pages>10-21</pages><issn>0303-7207</issn><eissn>1872-8057</eissn><abstract>•hCG induces CXCR4 expression in equine and bovine preovulatory follicles.•The hCG-dependent induction of CXCR4 occurs in both granulosa and theca cells.•Forskolin induces CXCR4 expression in bovine granulosa cell cultures.•Numerous signaling pathways are involved in follicular CXCR4 expression.•SDF1/CXCR4 activation increases the expression of ovulation-regulated genes.
The interaction between stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) has been implicated in leukocyte attraction, tissue remodeling and angiogenesis. The objective of the present study was to characterize the expression and regulation of SDF1 and CXCR4 in equine follicles during the ovulatory process. Equine preovulatory follicles were isolated during estrus 0–39h after hCG treatment. Follicle wall preparations (theca interna with attached granulosa cells) and isolated preparations of granulosa cells and theca interna were obtained, and total RNA extracts were analyzed by RT-PCR/Southern blot. Results showed that levels of CXCR4 transcripts were induced by hCG in follicles at 36h post-hCG (P<0.05 vs 0h), with the induction observed in both granulosa and theca cells. Immunoblotting and immunohistochemical analyses confirmed an increase in CXCR4 protein in follicles after hCG treatment. In contrast, levels of SDF1 transcripts were very low in granulosa cells but high in theca interna cells throughout most of the ovulatory period. Studies in vivo performed with bovine preovulatory follicles collected 0–24h post-hCG revealed a marked and significant up-regulation of CXCR4 transcripts after hCG (P<0.05), as observed in equine follicles. A similar pattern of CXCR4 mRNA up-regulation was observed in cultures of bovine granulosa cells treated with forskolin (P<0.05). This forskolin-dependent induction of CXCR4 mRNA was suppressed by co-treatment with inhibitors of PKA, ERK1/2 and EGFR, and by the progesterone receptor antagonist RU486 (P<0.05), underscoring the contribution of multiple signaling pathways. In complementary studies, treatment of bovine granulosa cells with EGF or the hypoxia mimetic cobalt chloride significantly increased CXCR4 transcript levels, whereas co-treatment with forskolin and a CXCR4 antagonist repressed the expression of several ovulation-related genes. Collectively, this study describes for the first time the gonadotropin-dependent up-regulation of CXCR4 transcript in ovarian follicles of large monoovulatory species, provides some insights into the regulation of CXCR4 gene expression in granulosa cells, and identifies a potential link between follicular SDF1/CXCR4 activation and the regulation of ovulation-related genes.</abstract><cop>Ireland</cop><pub>Elsevier Ireland Ltd</pub><pmid>24784705</pmid><doi>10.1016/j.mce.2014.04.009</doi><tpages>12</tpages></addata></record> |
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| ispartof | Molecular and cellular endocrinology, 2014-06, Vol.391 (1-2), p.10-21 |
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| subjects | Activation Amino Acid Sequence And ovulation-related genes Animals Cattle Cells, Cultured Cellular Chemokine CXCL12 - genetics Chemokine CXCL12 - metabolism Chemokine receptor Chlorides Chorionic Gonadotropin - administration & dosage Cobalt - pharmacology Colforsin - pharmacology Control Epidermal Growth Factor - pharmacology Estrus - physiology Female Gene expression Gene Expression Regulation Genes Gonadotropin Granulosa Cells - cytology Granulosa Cells - drug effects Granulosa Cells - metabolism Horses Humans Inhibitors Mifepristone - pharmacology Molecular Sequence Data Ovulation - physiology Ovulatory process Preovulatory follicles Primary granulosa cells Protein Kinase Inhibitors - pharmacology Receptors Receptors, CXCR4 - agonists Receptors, CXCR4 - antagonists & inhibitors Receptors, CXCR4 - genetics Receptors, CXCR4 - metabolism RNA, Messenger - agonists RNA, Messenger - antagonists & inhibitors RNA, Messenger - genetics RNA, Messenger - metabolism Signal Transduction Theca Cells - cytology Theca Cells - drug effects Theca Cells - metabolism |
| title | Expression and regulation of stromal cell-derived factor-1 (SDF1) and chemokine CXC motif receptor 4 (CXCR4) in equine and bovine preovulatory follicles |
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