渔业科学进展
漁業科學進展
어업과학진전
MARINE FISHERIES RESEARCH
2014年
4期
22-31
,共10页
柳学周%臧坤%徐永江%李春广%史宝%王妍妍
柳學週%臧坤%徐永江%李春廣%史寶%王妍妍
류학주%장곤%서영강%리춘엄%사보%왕연연
漠斑牙鲆%LHβ亚基%卵巢发育周期%表达特性%血浆激素浓度
漠斑牙鲆%LHβ亞基%卵巢髮育週期%錶達特性%血漿激素濃度
막반아평%LHβ아기%란소발육주기%표체특성%혈장격소농도
Paralichthys lethostigma%LHβ subunit%Ovarian maturation cycle%Expression pattern%Plasma hormone levels
利用cDNA末端快速克隆(Rapid Amplification of cDNA Ends, RACE)方法获得了漠斑牙鲆(Paralichthys lethostigma)促黄体素(Luteinizing Hormone, LH)β亚基的cDNA全长序列,检测了LHβ亚基mRNA的组织表达水平,揭示了垂体、肝脏和卵巢中LHβ亚基mRNA在卵巢发育周期中的表达水平变化,利用酶联免疫技术(Enzyme-Linked Immuno Sorbent Assay, ELISA)测定了血浆LH和雌二醇(Estrodiol, E2)表达水平的变化。结果表明,漠斑牙鲆为卵巢非同步发育分批产卵性鱼类, LHβ亚基cDNA序列全长597bp,开放阅读框(Open reading frame, ORF)长438 bp,编码145个氨基酸,LHβ亚基mRNA具有广泛的组织分布特性。与肝脏和卵巢相比,垂体中LHβ亚基mRNA表达水平在卵巢发育各个阶段都有较高表达水平,在IV期表达水平迅速升高至较高水平并保持至VI期。卵巢中LHβ mRNA表达水平在V期达到峰值,而肝脏中LHβ mRNA表达水平在II期时达到峰值,在V期也有相对较高的表达水平。血浆LH和E2表达水平在卵巢发育周期中均呈现趋势一致的规律性变化。研究结果可为认识漠斑牙鲆生殖调控机制提供基础资料。
利用cDNA末耑快速剋隆(Rapid Amplification of cDNA Ends, RACE)方法穫得瞭漠斑牙鲆(Paralichthys lethostigma)促黃體素(Luteinizing Hormone, LH)β亞基的cDNA全長序列,檢測瞭LHβ亞基mRNA的組織錶達水平,揭示瞭垂體、肝髒和卵巢中LHβ亞基mRNA在卵巢髮育週期中的錶達水平變化,利用酶聯免疫技術(Enzyme-Linked Immuno Sorbent Assay, ELISA)測定瞭血漿LH和雌二醇(Estrodiol, E2)錶達水平的變化。結果錶明,漠斑牙鲆為卵巢非同步髮育分批產卵性魚類, LHβ亞基cDNA序列全長597bp,開放閱讀框(Open reading frame, ORF)長438 bp,編碼145箇氨基痠,LHβ亞基mRNA具有廣汎的組織分佈特性。與肝髒和卵巢相比,垂體中LHβ亞基mRNA錶達水平在卵巢髮育各箇階段都有較高錶達水平,在IV期錶達水平迅速升高至較高水平併保持至VI期。卵巢中LHβ mRNA錶達水平在V期達到峰值,而肝髒中LHβ mRNA錶達水平在II期時達到峰值,在V期也有相對較高的錶達水平。血漿LH和E2錶達水平在卵巢髮育週期中均呈現趨勢一緻的規律性變化。研究結果可為認識漠斑牙鲆生殖調控機製提供基礎資料。
이용cDNA말단쾌속극륭(Rapid Amplification of cDNA Ends, RACE)방법획득료막반아평(Paralichthys lethostigma)촉황체소(Luteinizing Hormone, LH)β아기적cDNA전장서렬,검측료LHβ아기mRNA적조직표체수평,게시료수체、간장화란소중LHβ아기mRNA재란소발육주기중적표체수평변화,이용매련면역기술(Enzyme-Linked Immuno Sorbent Assay, ELISA)측정료혈장LH화자이순(Estrodiol, E2)표체수평적변화。결과표명,막반아평위란소비동보발육분비산란성어류, LHβ아기cDNA서렬전장597bp,개방열독광(Open reading frame, ORF)장438 bp,편마145개안기산,LHβ아기mRNA구유엄범적조직분포특성。여간장화란소상비,수체중LHβ아기mRNA표체수평재란소발육각개계단도유교고표체수평,재IV기표체수평신속승고지교고수평병보지지VI기。란소중LHβ mRNA표체수평재V기체도봉치,이간장중LHβ mRNA표체수평재II기시체도봉치,재V기야유상대교고적표체수평。혈장LH화E2표체수평재란소발육주기중균정현추세일치적규률성변화。연구결과가위인식막반아평생식조공궤제제공기출자료。
The role of the endocrine pathway in the regulation of southern flounder Paralichthys lethostigma reproduction has been unclear. Here we aimed to explore the underlying mechanisms by investigating the expression pattern of luteinizing hormoneβ subunit (LHβ). Using samples isolated from pituitary ofP. we obtained the full-length cDNA sequence of LHβ with the RACE method. The spatial and temporal expression patterns ofLHβmRNA in different tissues during an ovarian maturation cycle were analyzed using the qPCR assay. The levels of plasma LH and E2during an ovarian maturation cycle were also measured using the ELISA method. Our results revealed an asynchronous oocyte development and batch spawning in southern flounder. The cDNA sequence of LHβhad a length of 597 bp and it encoded 145 amino acids. LHβ expression was detected in all examined tissues except for muscle, which implied it could be involved in multiple physiological functions in southern flounder. The LHβ mRNA level in pituitary was high through the entire ovarian maturation cycle, and peaked at stage V and VI. In the ovary the LHβ mRNA level peaked at stage V. In the liver the LHβ mRNA level was the highest at stage II and relatively high at stage V. The levels of plasma LH and E2 fluctuated in a similar pattern during the ovarian maturation cycle. These results suggested that LHβmight regulate the ovarian maturation cycle via endocrine, paracrine and autocrine pathways. Our study provided insights into the functions of the endocrine pathway in the reproduction of southern flounder.