Expression pattern, cellular localization and promoter activity analysis of ovarian aromatase (Cyp19a1a) in protogynous hermaphrodite red-spotted grouper

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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
刊名	MOLECULAR AND CELLULAR ENDOCRINOLOGY
	2009-08-13
卷号	307 期号:1-2 页码:224-236
关键词	Immunofluorescene localization Promoter activity Cyp19a1a/b Grouper Hermaphrodite Sex reversal
ISSN号	0303-7207
通讯作者	Gui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
中文摘要	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.
英文摘要	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.
学科主题	Cell Biology; Endocrinology & Metabolism
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Cell Biology ; Endocrinology & Metabolism
研究领域[WOS]	Cell Biology ; Endocrinology & Metabolism
关键词[WOS]	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
收录类别	SCI
资助信息	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
语种	英语
WOS记录号	WOS:000267837400029
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/7654]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
作者单位	Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
推荐引用方式 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.(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.