三疣梭子蟹精子顶体反应前后胞内Ca^(2+)的变化  

Changes in intracellular Ca^(2+)of sperm fore-and-aft acrosome reaction in the swimming crab Portunus trituberculatus

作　　者：朱冬发[1] 沈丽萍[1] 沈建明[1] 吴琼[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,浙江宁波315211

出　　处：《动物学报》2008年第5期885-889,共5页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金(No.40576068);浙江省科技计划(No.2008C22048);浙江省自然科学基金(No.Y305339)资助~~

摘　　要：应用激光扫描共聚焦显微镜(LSCM)和Fluo-3/AM荧染技术对三疣梭子蟹精子顶体反应前后的胞内Ca2+变化进行了观察和检测。结果显示,在精子顶体反应过程中,胞内Ca2+主要分布在细胞核、穿孔器和胞质膜残存处,胞内Ca2+浓度([Ca2+]i)总体上呈现先上升后下降的趋势。顶体反应前精子的平均荧光强度为35.95±5.71;穿孔器前伸、顶体囊膜翻转阶段精子的平均荧光强度为66.80±7.35;顶体囊膜脱落、顶体丝形成阶段精子的平均荧光强度为3.87±2.82;上述各阶段间精子荧光强度有极显著差异(P<0.01)。顶体反应穿孔器前伸、顶体囊膜翻转阶段的精子相比顶体反应前精子,[Ca2+]i显著提高;而在顶体囊膜脱落、顶体丝形成阶段,[Ca2+]i则急剧下降,只在顶体丝基部胞质膜残存处有微量Ca2+存在。初步探讨了三疣梭子蟹精子顶体反应前后胞内Ca2+变化的功能。The acroseme reaction process of sperm in the swimming crab Portunus trituberculatus （Miers） consists of four consecutive stages： （1） swelling of the apical cap; （2） eversion of the acrosomal vesicle; （3） forward elongation of the perforatorium and reversal of the acrosomal vesicle membrane; and （4） shedding of the acrosomal vesicle membrane and formation of the acrosomal filament. In the present study, Laser Scanning Confocal Microscope （LSCM） was used to image spermatozoa stained with Fluo-3/AM at different stages of the fore-and-aft acrosome reaction. Fluo-3/AM was used as the Ca^2＋. fluorescence indicator. The 488 nm argon laser was used for excitation and the emitted fluorescence was 526 nm. On account of the stability of sperm during the acrosome reaction, we selected sperm at three stages （pre-acrosome reaction, the 3rd stage and the 4th stage of the acrosome reaction） to analyze intracellular Ca^2＋ changes. The average fluorescence intensity of sperms in the above-chosen stages were 35.95 ± 5.71, 66.80 ± 7.35 and 3.87 ±2.82 respectively. Average fluorescence intensities among these stages exhibited significant differences （ P 〈 0.01 ）. The intracellular calcium concentration （ [ Ca^2 ＋ ]i ） of sperm shows a tendency to rise first and then decrease during the acrosome reaction. The [Ca^2＋ ]i of the sperm in the 3rd stage of the acrosome reaction increased significantly compared with that of the sperm before acrosome reaction. In the 4th stage of the acrosome reaction, with the acresomal vesicle membrane shed and the acrosomal filament formed, the [ Ca^2＋ ]i of the sperm decreased sharply. The primary distribution sites of the intraccllular Ca^2＋ are the nucleus, perforatorium and cytoplasmic membrane remnants. The physiological actions of the intracellular Ca^2＋ changes of sperm during the fore-and-aft acrosome reaction in P. trituberculatus are also discussed

关 键 词：三疣梭子蟹 精子 顶体反应 激光扫描共聚焦显微镜 胞内CA^2+ 

分 类 号：Q954.4[生物学—动物学]