Evolutionary and ecological studies on the circum-north Pacific elements, the super-family cottoid

为了验证Cottoid内交配的多系进化假说，我们收集了一些鱼类样本，包括西太平洋物种和Cottoid物种生殖道的精细结构形态，以研究其分子遗传学。通过2006年3月在普吉湾和范库弗、2006年7月在乌纳拉斯卡岛和2007年11月在彼得大帝湾的3次考察，共采集到5科60种1500个标本。杜父鱼雌雄间的泌尿生殖投射形态差异出现在性腺分化完成后相当晚的时期，精巢和精子的形态在杜父鱼科中具有明显的特化。特别是在交配杜父鱼中发现了一些共同的特征，如精子头部细长、曙红阳性分泌、精子束、精子发生不同步等。本文以吉尔伯蒂半鳞鱼（Hemilepidotus gilberti）为材料，研究了生殖细胞向正常可育精子（真精子）和非可育双形精子（准精子）的分化。我们从睾丸中收集精母细胞，通过细胞培养获得了真精子和旁精子，表明DNA复制精母细胞能够自主进行精子发生的一部分以形成旁精子。通过对真精子和准精子酸提取液的电泳分析，在其电泳图谱中检测到了每种类型的特异性蛋白条带，表明不同类型精子的核浓缩是通过不同的精子特异性核碱性蛋白发生的。

To verify the polyphyletic evolution hypothesis of copulating within cottoid, we had the researches to collection of fish samples for investigation of molecular phylogeny including the western Pacific species and the fine structural morphology of the reproductive tract on cottoid species. We collected 5 families 60 species 1500 specimens during three expeditions, the Puget sounds and Vancuver in March 2006, Unalaska island in July 2006, and Peter the Great Bay in November 2007.Process of gonadal sex differentiation and formation of reproductive organ were investigated in uncopulative sculpin Cottus. In Cottus, the differences in morphology of urogenital projection between male and female appeared considerably late period after having completed gonadal sex differentiation.Remarkable specialization was admitted in the morphology of testis and sperm among Cottidae. Especially, some common characteristics (slender sperm head, eosin positive-secretion, sperm bunch, asynchronous spermatogenesis) were observed overcoming genus in copulative sculpins. It is thought that each characteristic is an adaptation to the copulation related to the spermatogenesis and the sperm motility.We investigated the differentiation of germ cells toward normal fertile sperm (eusperm) and non-fertile dimorphic sperm (parasperm) by using Hemilepidotus gilberti. We obtained eusperm and parasperm by cell culture of germ cells from spermatocytes collected from teites, indicating DNA-duplicating spermatocytes are potent to autonomously progress a part of spermatogenesis to form parasperm. By electrophoretic analysis of acid extracts from eusperm and parasperm, specific protein bands for each type were detected in its electrophoretic patterns, indicating the condensation of nucleus of the different type of sperm occur through a different sperm-specific nuclear basic proteins.