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|Publication Year :||1997|
以含有14C-leucine 的培養液對不同卵巢發育時期之卵巢及肝胰臟進行體外培養，培養8小時後測定各組織對14C-leucine的滲入量。卵巢及肝胰臟中蛋白質的合成速率均隨卵巢發育而降低，而肝胰臟中蛋白質的合成速率較卵巢為高。培養後之組織萃取後經電泳、放射顯影、西方點墨法檢驗後發現：卵巢可合成卵黃素之三個次單元體，肝胰臟合成可為抗卵黃素血清辨識之蛋白質均為較卵黃素之三個次單元體為小。唯一可見之較大分子之分子量約為 85kDa，與血藍素之次單元體之分子量十分接近。組織萃液以抗卵黃素血清及Protein A進行免疫沉澱，卵巢中新合成蛋白質中可為抗卵黃素血清辨識的百分比為3.7-11.0%，肝胰臟則佔2.4-14.0％。每毫克組織新合成蛋白質中可為抗卵黃素血清辨識之蛋白質放射量在卵巢為42-199dpm/mg protein，在肝胰臟則為54-440dpm/mg protein；兩者均隨卵巢發育而降低，且在卵巢發育各時期中肝胰臟卵黃素的合成量均高於卵巢。
Changes in the level of vitellin/vitellogenin(Vn/Vg) in the hepatopancreas and hemolymph are closely associated with ovarian maturation in the mud crab, Scylla serrata. Thus, it is possible that hepatopancreas is the extraovarian tissue of Vg synthesis. The study will provide evidence by in vitro culture.
Ovary and hepatopancreas from crab at different ovarian stages (oocyte diameter from 167 to 283μm) were incubated in MEM medium containing 14C-leucine. The incorporation of 14C-leucine into tissue was counted 8 hours after culture. The rate of protein synthesis in ovary and hepatopancreas decrease with ovarian maturation. And the rate of protein synthesis in hepatopancreas is higher than that in ovary. Tissue extracts from 8 hours incubation were analysis by electrophoresis, autoradiography, TCA percipitation and immunopercipitation. Ovary synthesize three subunits of Vn. Those proteins that are synthesized by hepatopancreas and can be recognized by anti-vitellin serum, are smaller than the subunits of Vn/Vg. The molecular weight of the only visiable band in autoradiography is about 85kDa, close to the molecular weight of hemocyanin subunits. The rates of protein synthesis are 42-199 dpm/mg protein in ovary and 54-440 dpm/mg protein in hepatopancreas, both decrease with ovarian maturation. Specific Vn/Vg synthesis was determined by immunoprecipitation with polyclonal Vn-specific antiserum and protein A. The percentage of Vn/Vg specific synthesis in the total protein synthesis are 3.7-11.0% in ovary and 2.4-11.0% in hepatopancreas. Judging the volum of the two organs, Vn/Vg synthesis in hepatopancreas are much higher than in ovary during those ovarian stages studied.
The results led to the following conclusions: 1). Both ovary and hepatopancreas are the sites of Vn synthesis in Scylla serrala. 2). Hepatopancreas is the major source of Vn/Vg, especially in the early stage of ovarian maturation. 3). The rate of protein synthesis in ovary and hepatopancreas decrease with ovarian maturation. 4). The percentage of Vn/Vg specific protein synthesis increased with ovarian maturation in ovary but decrease in hepatopancreas.
|Appears in Collections:||動物學研究所|
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