Molecular Mechanism of Anti-Mullerian Hormone in the Sex Differentiation and Sex Change of Protandrous Black Porgy

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Project title

Code/計畫編號

NSC99-2628-B019-005-MY3

Translated Name/計畫中文名

黑鯛性別分化及性轉變的分子遺傳機制：anti-mullerian hormone的角色

Project Coordinator/計畫主持人

Ching-Fong Chang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Aquaculture

Website

https://www.grb.gov.tw/search/planDetail?id=2380961

Year

2012

Start date/計畫起

01-08-2012

Expected Completion/計畫迄

31-07-2013

Bugetid/研究經費

1965千元

ResearchField/研究領域

漁業
生物技術(農)

"黑鯛第一年與第二年為雄性，第三年部分魚將性轉變為雌性，且以含雌二醇（estradiol, E2）飼料投餵4個月會誘導成為雌性，停止投餵後會導致變回雌性（reversible sex change），因此0-1歲全雄性黑鯛提供為探討性別分化之模式魚種，1-3年齡黑鯛則作為探討性轉變之魚種。本研究之目的以組織學、內分泌、細胞生物、外科切除技術及分子遺傳基因之層次，探討anti-mullerian hormone（Amh）在黑鯛生殖腺發育包括性別分化與性轉變之分子機制。實驗期間為三年，分別進行下列之實驗： 1) 瞭解黑鯛性別分化與性轉變過程，生殖管（genital ducts）之發育特性。 2) 探討3個月齡至1年齡期間雄性黑鯛性別分化相關重要基因（Amh及Amh receptor）之關係。 3) 以E2誘導性轉變後，再變為雌性為模式（reversible sex change），探討對雄性（Amh及Amh receptor）與雌性（Fig α與cyp19a1a）相關重要基因的關係。 4) 以in situ hybridization方法確定Amh及Amh receptor在雄性生殖腺分化所表現的位置。 5) 探討Amh及Amh receptor在黑鯛1至3年齡之季節性變化與性轉變之關係。 6) 以外科手術技術（切除精巢或切除其中一邊之生殖腺），探討卵巢與精巢Amh及Amh receptor在黑鯛性轉變過程之可能角色。 7) 以酵母菌表現Amh蛋白，進而以精巢與卵巢進行組織培養（in vitro tissue culture）的技術及添加Amh，探討Amh對精巢與卵巢組織的發育作用。 8) 以Amh RNA interference（基因弱化）的技術，探討Amh在黑鯛生殖腺發育與性轉變之角色。經由本計畫之執行，預期瞭解生殖腺Amh及Amh receptor之表現是否與黑鯛性別分化與性轉變有關係，進而可說明Amh對精巢與卵巢發育之影響及其可能的作用機制。" "Black porgy, Acanthopagrus schlegeli Beeker, a marine protandrous hermaphrodite, is functional male for the first 2 years of life but begin to sexually change to female after the third year. Estradiol (E2) could induce a reversible sex change in 1-year-old fish.This sex pattern provides an unique model to study the molecular mechanism of male sex differentiation and sex change in gonad. The objectives are to investigate the molecular mechanism of gonadal sex differentiation in protandrous black porgy in the relation to the expression and function of Amh (anti-mullerian hormone) and Amh receptor. The experiments will include as follows: 1) The expression profiles of Amh , Amh receptor and vasa (germ cell marker) transcripts in gonad during gonadal sex differentiation in black progy. 2) The localization of Amh and Amh receptor gene in the differentiating gonad by in situ hybridization. 3) The expression profiles of male genes (Amh and Amh receptor trascripts), female gene (Fig α and cyp19a1a) and vasa (germ cell marker) in the gonad of E2-induced (fed) females. 4) The expression profiles of germ cell marker (vasa), male genes (Amh and Amh receptor transcripts), and female gene (Fig α and cyp19a1a) during reversible sex chage from E2-induced female to a male. 5) The expression profile of Amh and Amh receptor during the seasonal change and natural sex change in 1- to 3-year-old black porgy. 6) The expression profile of Amh and Amh receptor in the gonadal tissue after surgically removal of testicular tissue or one of the pair gonads. 7) The effect of Amh in the development and cellular activity of gonadal tissue by tissue culture. 8) The specific roles of Amh in the gonadal (testicular and ovarian development) by Amh RNA interference technique. 9) The specific effects of Amh in the development of testicular and ovarian tissue in the bisexual gonad. We anticipate to obtain the significant and novel results and explain the function and mechanism of anti-mullerian hormone in the development of gonad."

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Molecular Mechanism of Anti-Mullerian Hormone in the Sex Differentiation and Sex Change of Protandrous Black Porgy

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