title: Calcineurin and CRTC2 mediate FSH and TGFβ1 upregulation of Cyp19a1 and Nr5a in ovary granulosa cells.
abstract: Estrogens are essential for female reproduction and overall well-being, and estrogens in the circulation are largely synthesized in ovarian granulosa cells. Using primary cultures of ovarian granulosa cells from gonadotropin-primed immature rats, we have recently discovered that pituitary FSH and ovarian cytokine transforming growth factor beta 1 (TGFβ1) induce calcineurin-mediated dephosphorylation-activation of cAMP-response element-binding protein (CREB)-regulated transcription coactivator (CRTC2) to modulate the expression of Star, Cyp11a1, and Hsd3b leading to increased production of progesterone. This study explored the role of calcineurin and CRTC2 in FSH and TGFβ1 regulation of Cyp19a1 expression in granulosa cells. Ovarian granulosa cells treated with FSH displayed increased aromatase protein at 24  h post-treatment, which subsided by 48  h, while TGFβ1 acting through its type 1 receptor augmented the action of FSH with a greater and longer effects. It is known that the ovary-specific Cyp19a1 PII-promoter contains crucial response elements for CREB and nuclear receptor NR5A subfamily liver receptor homolog 1 (LRH1/NR5A2) and steroidogenic factor 1 (SF1/NR5A1), and that the Nr5a2 promoter also has a potential CREB-binding site. Herein, we demonstrate that FSH+TGFβ1 increased LRH1 and SF1 protein levels, and their binding to the Cyp19a1 PII-promoter evidenced, determined by chromatin immunoprecipitation analysis. Moreover, pretreatment with calcineurin auto-inhibitory peptide (CNI) abolished the FSH+TGFβ1-upregulated but not FSH-upregulated aromatase activity at 48  h, and the corresponding mRNA changes in Cyp19a1, and Nr5a2 and Nr5a1 at 24  h. In addition, FSH and TGFβ1 increased CRTC2 binding to the Cyp19a1 PII-promoter and Nr5a2 promoter at 24  h, with CREB bound constitutively. In summary, the results of this study indicate that calcineurin and CRTC2 have important roles in mediating FSH and TGFβ1 collateral upregulation of Cyp19a1 expression together with its transcription regulators Nr5a2 and Nr5a1 in ovarian granulosa cells.
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan.
authors: Wei-An
authors: WA
authors: Lai
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan.
authors: Yi-Ting
authors: YT
authors: Yeh
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan.
authors: Wei-Ling
authors: WL
authors: Fang
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan.
authors: Leang-Shin
authors: LS
authors: Wu
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan.
authors: Nobuhiro
authors: N
authors: Harada
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan.
authors: Peng-Hui
authors: PH
authors: Wang
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan jiuanh@ym.edu.tw johnweiwang@hotmail.com fck@ccms.ntu.edu.tw.
authors: Ferng-Chun
authors: FC
authors: Ke
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan jiuanh@ym.edu.tw johnweiwang@hotmail.com fck@ccms.ntu.edu.tw.
authors: Wen-Ling
authors: WL
authors: Lee
authors: Institute of PhysiologySchool of Medicine, National Yang-Ming University, 155 Linong Street, Section 2, Taipei 11221, TaiwanDepartment of NursingHsin-Sheng College of Medical Care and Management, Taoyuan, TaiwanDepartment of Animal Science and TechnologyCollege of Bio-Resources and Agriculture, National Taiwan University, Taipei, TaiwanDepartment of BiochemistrySchool of Medicine, Fujita Health University, Aichi, JapanDepartment of Obstetrics and GynecologyTaipei Veterans General Hospital, Taipei, TaiwanDepartment of Obstetrics and GynecologyNational Yang-Ming University, Taipei, TaiwanInstitute of Molecular and Cellular BiologyCollege of Life Science, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei 10617, TaiwanDepartment of MedicineCheng Hsin General Hospital, 45 Jhensing Street, Taipei 11220, TaiwanDepartment of NursingOriental Institute of Technology, New Taipei City, Taiwan jiuanh@ym.edu.tw johnweiwang@hotmail.com fck@ccms.ntu.edu.tw.
authors: Jiuan-Jiuan
authors: JJ
authors: Hwang
mesh_terms: D000818:Animals; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D019703:Calcineurin / Q000378:metabolism* / Q000494:pharmacology; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism* / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D054339:Steroidogenic Factor 1 / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism* / Q000494:pharmacology; D015854:Up-Regulation
chemical_list: C545700:CRTC2 protein, rat; D054339:Steroidogenic Factor 1; D015534:Trans-Activators; D053773:Transforming Growth Factor beta1; D005640:Follicle Stimulating Hormone; D001141:Aromatase; D019703:Calcineurin
keywords: CRTC2; FSH; LRH1/Nr5a2; SF1/Nr5a1; TGFβ; aromatase/Cyp19a1; calcineurin; granulosa cell; ovary