动物学报
動物學報
동물학보
ACTA ZOOLOGICA SINICA
2001年
1期
79-84
,共6页
雌核发育%精子蛋白%精头膜%鞭毛%脱膜精头%分级分离
雌覈髮育%精子蛋白%精頭膜%鞭毛%脫膜精頭%分級分離
자핵발육%정자단백%정두막%편모%탈막정두%분급분리
对雌核发育银鲫和两性融合彩鲫精子蛋白组份进行了比较分析。通过分级抽提得到精子的不同组份精浆、精头的膜、鞭毛和脱膜精头等,然后经不同的凝胶电泳系统,比较分析了银鲫精子和其两性亲缘种彩鲫精子相应组份可溶性蛋白成份的差异。研究表明,经分级抽提的银鲫精子和彩鲫精子的各个组份都含有其特定的蛋白谱带。精浆蛋白在两种鱼之间和两种鱼的不同个体之间都存在一定差异。精头膜、鞭毛和脱膜精头的可溶性蛋白在同种鱼不同个体间高度一致,但在两种鱼之间表现出差异。两种鱼精头膜的可溶性蛋白在SDS-PAGE电泳图谱上基本一致,而在非变性聚丙烯酰胺凝胶电泳图谱上则具有各自的特征性谱带。鞭毛可溶性蛋白的SDS-PAGE分析在雌核发育银鲫中揭示出一条特异的蛋白带。脱膜精头的可溶性蛋白在SDS-PAGE电泳图谱上差异明显,存在几条特征性蛋白带,并经Acid-Urea PAGE系统分析,证实这些特征性蛋白为碱性蛋白。这些发现为进一步鉴定雌核发育银鲫雄鱼精子的特异性蛋白和揭示其分子机制打下了基础。
對雌覈髮育銀鯽和兩性融閤綵鯽精子蛋白組份進行瞭比較分析。通過分級抽提得到精子的不同組份精漿、精頭的膜、鞭毛和脫膜精頭等,然後經不同的凝膠電泳繫統,比較分析瞭銀鯽精子和其兩性親緣種綵鯽精子相應組份可溶性蛋白成份的差異。研究錶明,經分級抽提的銀鯽精子和綵鯽精子的各箇組份都含有其特定的蛋白譜帶。精漿蛋白在兩種魚之間和兩種魚的不同箇體之間都存在一定差異。精頭膜、鞭毛和脫膜精頭的可溶性蛋白在同種魚不同箇體間高度一緻,但在兩種魚之間錶現齣差異。兩種魚精頭膜的可溶性蛋白在SDS-PAGE電泳圖譜上基本一緻,而在非變性聚丙烯酰胺凝膠電泳圖譜上則具有各自的特徵性譜帶。鞭毛可溶性蛋白的SDS-PAGE分析在雌覈髮育銀鯽中揭示齣一條特異的蛋白帶。脫膜精頭的可溶性蛋白在SDS-PAGE電泳圖譜上差異明顯,存在幾條特徵性蛋白帶,併經Acid-Urea PAGE繫統分析,證實這些特徵性蛋白為堿性蛋白。這些髮現為進一步鑒定雌覈髮育銀鯽雄魚精子的特異性蛋白和揭示其分子機製打下瞭基礎。
대자핵발육은즉화량성융합채즉정자단백조빈진행료비교분석。통과분급추제득도정자적불동조빈정장、정두적막、편모화탈막정두등,연후경불동적응효전영계통,비교분석료은즉정자화기량성친연충채즉정자상응조빈가용성단백성빈적차이。연구표명,경분급추제적은즉정자화채즉정자적각개조빈도함유기특정적단백보대。정장단백재량충어지간화량충어적불동개체지간도존재일정차이。정두막、편모화탈막정두적가용성단백재동충어불동개체간고도일치,단재량충어지간표현출차이。량충어정두막적가용성단백재SDS-PAGE전영도보상기본일치,이재비변성취병희선알응효전영도보상칙구유각자적특정성보대。편모가용성단백적SDS-PAGE분석재자핵발육은즉중게시출일조특이적단백대。탈막정두적가용성단백재SDS-PAGE전영도보상차이명현,존재궤조특정성단백대,병경Acid-Urea PAGE계통분석,증실저사특정성단백위감성단백。저사발현위진일보감정자핵발육은즉웅어정자적특이성단백화게시기분자궤제타하료기출。
In order to reveal the difference of sperm proteins between males from gynogenetic Silver crucian carp (Carassius auratus gibelio) and gonochoristic transparent Color crucian carp (Carassius auratus transparent colored varity), the two types of sperm were respectively extracted by the same methods at same conditions, and various fractions were collected. The fractions include sperm plasma, sperm head membrane, flagella, and demembranated sperm head.
The soluble protein components of these extracted fractions and the whole sperm were analysed by means of different polyacrylamide gel electrophoresis system, such as nondenaturing polyacrylamide gel electrophoresis, SDS-PAGE and Acid-Urea PAGE system. The obtained results show that each fraction contains specific soluble protein components, and a few of soluble proteins are different in some fraction between gynogenetic Silver crucian carp and gonochoristic Color crucian carp. The protein components of sperm plasma are different not only between the two kinds of fishes, but also among the individuals in the same kind of fish. It is suggested that the difference of sperm plasma proteins could be resulted from the contamination of blood and body fluid.
In the flagella fraction, one soluble protein component specific for gynogenetic Silver crucain carp was observed by SDS-PAGE system. The soluble protein components in sperm head membrane fraction were analysed by SDS-PAGE system and nondenaturing PAGE system, and found that they have same patterns in SDS-PAGE system but various pattern in nondenaturing PAGE system. It was suggested that the difference might be related to complex and functional status of the head membrane proteins. The significant difference of the soluble protein components between two types of fishes were discovered in demembranated sperm head fraction. Several protein bands specific for gynogenetic Silver crucain carp were revealed by SDS-PAGE analysis. Furthermore, the specific proteins were demonstrated to be basic proteins by Acid-Urea PAGE system. These discoveries laid a foundation for identifying specific proteins and revealing the mloe cular mechanism in gynogenetic Silver crucian carp sperm