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Expression pattern, cellular localization and promoter activity analysis of ovarian aromatase (Cyp19a1a) in protogynous hermaphrodite red-spotted grouper
Huang, Wei; Zhou, Li; Li, Zhi; Gui, Jian-Fang; Gui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2009-08-13
Source PublicationMOLECULAR AND CELLULAR ENDOCRINOLOGY
ISSN0303-7207
Volume307Issue:1-2Pages:224-236
AbstractAromatase plays a key role in sex differentiation of gonads. In this study, we cloned the full-length cDNA of ovarian aromatase from protogynous hermaphrodite red-spotted grouper (Epinephelus akaara), and prepared the corresponding anti-EaCyp19a1a antiserum. Western blot and immunofluorescence studies revealed ovary-specific expression pattern of EaCyp19a1a in adults and its dynamic expression change during artificial sex reversal. EaCyp19a1a was expressed by follicular cells of follicular layer around oocytes because strong EaCyp19a1a immunofluorescence was observed in the cells of ovaries. During artificial sex reversal, EaCyp19a1a expression dropped significantly from female to male, and almost no any positive EaCyp19a1a signal was observed in testicular tissues. Then, we cloned and sequenced a total of 1967 bp T-flanking sequence of EaCyp19a1a promoter, and showed a number of potential binding sites for some transcriptional factors, such as SOX5, GATA gene family, CREB, AP1, FOXL1, C/EBP, ARE and SF-1. Moreover, we prepared a series of 5' deletion promoter constructs and performed in vitro luciferase assays of EaCyp19a1a promoter activities. The data indicated that the CREB regulation region from -1010 to -898 might be a major cis-acting element to EaCyp19a1a promoter, whereas the elements GATA and SOX5 in the region from -1216 to -1010 might be suppression elements. Significantly, we found a common conserved sequence region in the fish ovary-type aromatase promoters with identities from 93% to 34%. And, the motifs of TATA box, SF-1, SOX5, and CREB existed in the region and were conserved among the most of fish species. (C) 2009 Elsevier Ireland Ltd. All rights reserved.; Aromatase plays a key role in sex differentiation of gonads. In this study, we cloned the full-length cDNA of ovarian aromatase from protogynous hermaphrodite red-spotted grouper (Epinephelus akaara), and prepared the corresponding anti-EaCyp19a1a antiserum. Western blot and immunofluorescence studies revealed ovary-specific expression pattern of EaCyp19a1a in adults and its dynamic expression change during artificial sex reversal. EaCyp19a1a was expressed by follicular cells of follicular layer around oocytes because strong EaCyp19a1a immunofluorescence was observed in the cells of ovaries. During artificial sex reversal, EaCyp19a1a expression dropped significantly from female to male, and almost no any positive EaCyp19a1a signal was observed in testicular tissues. Then, we cloned and sequenced a total of 1967 bp T-flanking sequence of EaCyp19a1a promoter, and showed a number of potential binding sites for some transcriptional factors, such as SOX5, GATA gene family, CREB, AP1, FOXL1, C/EBP, ARE and SF-1. Moreover, we prepared a series of 5' deletion promoter constructs and performed in vitro luciferase assays of EaCyp19a1a promoter activities. The data indicated that the CREB regulation region from -1010 to -898 might be a major cis-acting element to EaCyp19a1a promoter, whereas the elements GATA and SOX5 in the region from -1216 to -1010 might be suppression elements. Significantly, we found a common conserved sequence region in the fish ovary-type aromatase promoters with identities from 93% to 34%. And, the motifs of TATA box, SF-1, SOX5, and CREB existed in the region and were conserved among the most of fish species. (C) 2009 Elsevier Ireland Ltd. All rights reserved.
SubtypeArticle
KeywordImmunofluorescene Localization Promoter Activity Cyp19a1a/b Grouper Hermaphrodite Sex Reversal
Department[Huang, Wei; Zhou, Li; Li, Zhi; Gui, Jian-Fang] Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Subject AreaCell Biology ; Endocrinology & Metabolism
DOI10.1016/j.mce.2009.04.003
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology ; National Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology ; National Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology ; National Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology
Indexed BySCI
Language英语
WOS Research AreaCell Biology ; Endocrinology & Metabolism
WOS SubjectCell Biology ; Endocrinology & Metabolism
WOS IDWOS:000267837400029
WOS KeywordTILAPIA OREOCHROMIS-NILOTICUS ; MESSENGER-RNA EXPRESSION ; TROUT ONCORHYNCHUS-MYKISS ; BASS DICENTRARCHUS-LABRAX ; EUROPEAN SEA BASS ; P450 AROMATASE ; EPINEPHELUS-COIOIDES ; NILE TILAPIA ; DEVELOPMENTAL EXPRESSION ; CYTOCHROME-P450 AROMATASES
Funding OrganizationNational Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology ; National Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology ; National Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology ; National Major Basic Research [2004CB117401]; National Natural Science Foundation of China ; Guangdong Province [U0631007]; Chinese Academy of Sciences [KSCX2-YW-N-020, 075A011301]; Innovation Project of Institute of Hydrobiology
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Cited Times:57[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/7654
Collection期刊论文
Corresponding AuthorGui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Huang, Wei,Zhou, Li,Li, Zhi,et al. Expression pattern, cellular localization and promoter activity analysis of ovarian aromatase (Cyp19a1a) in protogynous hermaphrodite red-spotted grouper[J]. MOLECULAR AND CELLULAR ENDOCRINOLOGY,2009,307(1-2):224-236.
APA Huang, Wei,Zhou, Li,Li, Zhi,Gui, Jian-Fang,&Gui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2009).Expression pattern, cellular localization and promoter activity analysis of ovarian aromatase (Cyp19a1a) in protogynous hermaphrodite red-spotted grouper.MOLECULAR AND CELLULAR ENDOCRINOLOGY,307(1-2),224-236.
MLA Huang, Wei,et al."Expression pattern, cellular localization and promoter activity analysis of ovarian aromatase (Cyp19a1a) in protogynous hermaphrodite red-spotted grouper".MOLECULAR AND CELLULAR ENDOCRINOLOGY 307.1-2(2009):224-236.
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