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Molecular and expression characterization of three gonadotropin subunits common alpha, FSH beta and LH beta in groupers
Li, CJ; Zhou, L; Wang, Y; Hong, YH; Gui, JF; Gui, JF, Chinese Acad Sci, Grad Sch, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol,Inst Hydrobiol, Wuhan 430072, Peoples R China
2005-04-15
Source PublicationMOLECULAR AND CELLULAR ENDOCRINOLOGY
ISSN0303-7207
Volume233Issue:1-2Pages:33-46
AbstractA SMART cDNA plasmid library was constructed from protogyous greasy grouper (Epinephelus coioides) pituitary, and the full-length cDNAs of three gonadotropin (GTH) subunits common alpha, FSH beta and LH beta were cloned and sequenced from the library. The nucleotide sequences of common alpha, FSH beta and LH beta subunit cDNAs are 647, 594 and 574 bp in length, and encode for mature peptides of 94, 99 and 115 aa, respectively. High homology was observed by amino acid sequence alignment and identity comparison of the grouper mature peptides of common alpha, FSH beta and LH beta with that of other fishes. Phylogenetic tree analyses of the three GTH mature subunits revealed similar phylogeny relationships among the studied fish species. Three polyclonal antibodies were prepared from the in vitro expressed common alpha, FSH beta and LH beta mature proteins, respectively. Western blot analysis and immunofluoresence localization were performed on two typical stages of ovarian development stages in red-spotted grouper. Significant differences in protein expression levels of three gonadotropin subunits were revealed between the two ovarian development stages. In the individuals with resting ovary, common alpha was almost not detected in pituitaries, and FSH beta and LH beta expression levels were very low. While in the individuals with developing ovary, the expression of all three gonadotropin subunits reached to a high level. Immunofluoresence localization indicated that the grouper FSH beta cells mainly distributed in the middle area of PPD, while the LH beta cells distributed more widely, including in the area similar to the FSH beta cells and at the external periphery of pituitary near to the PI side. The common alpha might be expressed in both FSH beta and LH beta cells. Double immunofluoresence localization further demonstrated FSH beta and LH beta expression in distinct cells in the PPD area, although the FSH beta and LH beta cells were detected in the identical area of PPD. (c) 2005 Elsevier Ireland Ltd. All rights reserved.; A SMART cDNA plasmid library was constructed from protogyous greasy grouper (Epinephelus coioides) pituitary, and the full-length cDNAs of three gonadotropin (GTH) subunits common alpha, FSH beta and LH beta were cloned and sequenced from the library. The nucleotide sequences of common alpha, FSH beta and LH beta subunit cDNAs are 647, 594 and 574 bp in length, and encode for mature peptides of 94, 99 and 115 aa, respectively. High homology was observed by amino acid sequence alignment and identity comparison of the grouper mature peptides of common alpha, FSH beta and LH beta with that of other fishes. Phylogenetic tree analyses of the three GTH mature subunits revealed similar phylogeny relationships among the studied fish species. Three polyclonal antibodies were prepared from the in vitro expressed common alpha, FSH beta and LH beta mature proteins, respectively. Western blot analysis and immunofluoresence localization were performed on two typical stages of ovarian development stages in red-spotted grouper. Significant differences in protein expression levels of three gonadotropin subunits were revealed between the two ovarian development stages. In the individuals with resting ovary, common alpha was almost not detected in pituitaries, and FSH beta and LH beta expression levels were very low. While in the individuals with developing ovary, the expression of all three gonadotropin subunits reached to a high level. Immunofluoresence localization indicated that the grouper FSH beta cells mainly distributed in the middle area of PPD, while the LH beta cells distributed more widely, including in the area similar to the FSH beta cells and at the external periphery of pituitary near to the PI side. The common alpha might be expressed in both FSH beta and LH beta cells. Double immunofluoresence localization further demonstrated FSH beta and LH beta expression in distinct cells in the PPD area, although the FSH beta and LH beta cells were detected in the identical area of PPD. (c) 2005 Elsevier Ireland Ltd. All rights reserved.
SubtypeArticle
KeywordGonadotropins Common Alpha Fsh Beta Lh Beta Grouper Pituitary
DepartmentChinese Acad Sci, Grad Sch, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol,Inst Hydrobiol, Wuhan 430072, Peoples R China
Subject AreaCell Biology ; Endocrinology & Metabolism
DOI10.1016/j.mce.2005.01.010
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaCell Biology ; Endocrinology & Metabolism
WOS SubjectCell Biology ; Endocrinology & Metabolism
WOS IDWOS:000228166800004
WOS Keyword2 DISTINCT GONADOTROPINS ; FOLLICLE-STIMULATING-HORMONE ; MESSENGER-RNA LEVELS ; SALMONID PITUITARY GONADOTROPHS ; TROUT ONCORHYNCHUS-MYKISS ; EEL ANGUILLA-JAPONICA ; SEABREAM PAGRUS-MAJOR ; FEMALE RAINBOW-TROUT ; GTH-II ; GENE-EXPRESSION
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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/152342/9294
Collection期刊论文
Corresponding AuthorGui, JF, Chinese Acad Sci, Grad Sch, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol,Inst Hydrobiol, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, Grad Sch, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol,Inst Hydrobiol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Li, CJ,Zhou, L,Wang, Y,et al. Molecular and expression characterization of three gonadotropin subunits common alpha, FSH beta and LH beta in groupers[J]. MOLECULAR AND CELLULAR ENDOCRINOLOGY,2005,233(1-2):33-46.
APA Li, CJ,Zhou, L,Wang, Y,Hong, YH,Gui, JF,&Gui, JF, Chinese Acad Sci, Grad Sch, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol,Inst Hydrobiol, Wuhan 430072, Peoples R China.(2005).Molecular and expression characterization of three gonadotropin subunits common alpha, FSH beta and LH beta in groupers.MOLECULAR AND CELLULAR ENDOCRINOLOGY,233(1-2),33-46.
MLA Li, CJ,et al."Molecular and expression characterization of three gonadotropin subunits common alpha, FSH beta and LH beta in groupers".MOLECULAR AND CELLULAR ENDOCRINOLOGY 233.1-2(2005):33-46.
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