| 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 Publication | MOLECULAR AND CELLULAR ENDOCRINOLOGY
 |
| ISSN | 0303-7207 |
| Volume | 307Issue:1-2Pages:224-236 |
| Abstract | 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.; 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. |
| Subtype | Article |
| Keyword | Immunofluorescene 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 Area | Cell Biology ; Endocrinology & Metabolism |
| DOI | 10.1016/j.mce.2009.04.003 |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| Funding Organization | 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 ; 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 By | SCI |
| Language | 英语 |
| WOS Research Area | Cell Biology ; Endocrinology & Metabolism |
| WOS Subject | Cell Biology ; Endocrinology & Metabolism |
| WOS ID | WOS:000267837400029 |
| WOS Keyword | TILAPIA 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 Organization | 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 ; 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 |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ihb.ac.cn/handle/152342/7654 |
| Collection | 期刊论文 |
| Corresponding Author | Gui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| Affiliation | Chinese 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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