【作者】 陈礼强；

【导师】 郑曙明；

【作者基本信息】 西南大学 ， 水产养殖， 2007， 硕士

【摘要】 细鳞裂腹鱼(Schizothorax chongi)隶属于鲤科(Cyprinidae)裂腹鱼亚科(Schizothoracinae)，是产区的名贵鱼类，也是中国特有的重要冷水性经济鱼类。本研究以细鳞裂腹鱼为试验材料，研究了性腺发育成熟过程中各组织中营养物质的积累和转化及其相关酶活性的变化规律；通过组织切片和透射电镜技术对细鳞裂腹鱼性腺发育的显微和超微结构进行了观测；还对其人工繁育技术和胚胎发育特点进行了探讨。1细鳞裂腹鱼性腺成熟过程中体内的生化组成和相关酶活性测定雌雄细鳞裂腹鱼的性腺系数(GSI)均随性腺的成熟而逐渐升高，到第Ⅴ期达到最大值，分别为8.78±3.93和5.54±2.03；雄鱼的肝胰脏系数(HSI)在第Ⅴ期最高，为2.13±0.22；而雌鱼性腺的脂肪系数(CF)则在第Ⅳ~+期到达峰值14.61±3.44。对性腺发育成熟过程中肌肉、肝胰脏、性腺和血液中各项生理生化指标的检测表明：水分在各时期各组织中的变化较小；肌糖原随性腺成熟变化小，而肝糖原和血糖则呈现剧烈的变化；卵巢在发育的早期，对蛋白质的需求较大，而对脂类物质的需求不高，该期卵巢积累的蛋白质和脂类均来源于体内的其他组织；卵巢成熟后期对蛋白质的需求减少，但对脂类的需求急剧增加，在此阶段脂质主要是靠内源供应。精巢中乳酸脱氢酶(LDH)活性与其精子成熟的代谢密切相关，并随精子的成熟而升高；性腺中的脂肪酶(LPS)活性也随性腺发育成熟而逐渐升高；一氧化氮合酶(NOS)活性在性腺成熟前的各组织中均以“脉冲”的方式表达，可能是一种生殖信号。2细鳞裂腹鱼性腺发育的显微和超微结构研究系统地描述了各期精、卵巢的形态结构和各时相生殖细胞的显微和超微结构特征。将精、卵巢的发育都分为6个时期，相应的卵母细胞发育分为6个时相，生精细胞发育经历了初级精原细胞、次级精原细胞、初级精母细胞、次级精母细胞、精子细胞和成熟精子几个阶段。细鳞裂腹鱼精巢属于小叶型。精小叶边缘有各种非生殖细胞分布，包括支持细胞、边界细胞、成纤维细胞和间质细胞，各类生精细胞位于精小叶中部。初级精原细胞和次级精原细胞在精巢中的分布区域，细胞大小，细胞器种类和数量的分布上有诸多不同。拟染色体在除精子以外的各期生殖细胞中存在。核泡是在精子细胞形成的过程中产生的，并随着精子的成熟而消失。成熟精子头部无顶体，主要为核占据，尾部细长，主要由轴丝组成，呈“9+2”结构。卵巢是由生殖脊包裹着生长发育的各级卵母细胞构成。卵母细胞发育到第3时相末，卵黄开始在细胞核与细胞膜的中间部位积累，并围绕细胞核呈环状分布，随后逐渐往细胞核和细胞膜两边扩散。卵黄的形成经历了卵黄颗粒前体、卵黄颗粒中间体和卵黄颗粒3个阶段。在卵母细胞发育成熟过程中，核仁数目呈波浪状起伏，到第3时相晚期达到最多，为118±26个，核仁颗粒在第2时相晚期达到最大，直径为7.0±1.6μm。3细鳞裂腹鱼人工繁殖研究精子在天然河水中快速运动时间为43±8 s，寿命为180±36 s。精子在不同浓度的氯化钠和葡萄糖溶液中快速运动时间和寿命的变化规律基本一致。在0.6％的氯化钠溶液和0.4％的葡萄糖溶液中精子的活力最强，其精子快速运动的平均时间分别为54 s和52 s，平均寿命分别为1508s和1760s。加入去离子水后，停止运动的精子又可被激活。2005～2006年，分别用4种不同的催产剂组合对经人工驯养的180余尾性成熟的细鳞裂腹鱼进行了人工催产，共获得受精卵50余万粒，孵化出仔鱼9万余尾。可成功进行人工催产的水温范围为13～18℃。选择出催产效果较好的催产剂组合为：PG与HCG联合二次注射，第一次注射剂量为(5mg+600IU)/kg鱼体重，第二次注射剂量为(12mg+1000IU)/kg鱼体重，在水温13～18℃效应时间为85～98 h。在水温17±1℃的条件下，胚胎发育历时124 h后孵出仔鱼，孵出9d后仔鱼鳔充气并开始平游。从受精卵到孵化出膜所需要的平均积温为2108 h·℃。胚胎发育可分为6个阶段，共25个发育时期，各发育时期的特征均被描述。在裂腹鱼亚科中，细鳞裂腹鱼胚胎和仔鱼发育速度较快，在肌肉效应期以后更加明显；心跳频率并不是简单的由慢到快的线性增长，而是一个快慢相间的曲线变化的过程；胚胎发育的各个时期对环境的敏感性不同，原肠期对外界环境变化最为敏感。更多还原

【Abstract】 Schizothorax chongi, which belongs to Schizothoracinae, Cyprinidae, is one of the most important economical cold water fish, uniquely lived in China. In this paper, the reproductive biology of S. chongi was discussed. The results indicated as follows:1. The detection of biochemical composition and enzymes activities in vivo tissues of S.chongi with the gonad maturityGonadosomatic index (GSI) was gradually rising with the gonad maturity and achieved the maximum at the stage V, arriving at 8.78+3.93 and 5.54±2.03 respectively. Hepatopancreas index (HSI) of male also reached maximum of 2.13±0.22 at stage V, however, the female gonadal coefficient of fat (CF) in stage IV~+ reached the peak of 14.61±3.44.Physiological and biochemical indices were detected in different tissues during gonad development and maturation. The results show that: tissue water contents changes very little in different stages. Muscle glycogen has a small change, but liver glycogen and glucose showed a drastic change with the gonad maturity. In the early stages, ovarian demand for a great deal of protein but little lipid, the resource of protein and lipid accumulated in vivo Came from other organizations; In the later stages, mature ovarian reduced the demand for protein, but the demand for lipid increased dramatically, and the lipid mainly relies on internal supply at this stage; Testicular lactate dehydrogenase (LDH) activity was closely related to the sperm mature metabolism, and gradually increased with the mature sperm; Lipase (LPS) activity in gonads along with the gonad maturity and gradually increased; Nitric oxide synthase (NOS) activity in the organizations are "pulse" of expression before gonads mature, it may be a reproductive signal.2. Histological and untrastructural studies on the gonad development of S.chongiThe microstructure of the gonad development and the intrastructure of stem cells of S. chongi were systemic depicted. According to the morphological and histological features, both testis and ovary development in S. chongi can be divided into six stages, and the change of female cells can be divided into six phases. There are several phases in the spermatogenesis, namely are primary spermatogonium、secondary spermatogonium、primary spermatocytes、secondary spermatocytes、spermatid and mature sperm. The testis was lobular type. There are some somatic cells around the testis, such as sertoli cell and leyding cell, fibroblasts and so on. Various spermatogenic cells locate in the central lobule. Primary spermatogonia and secondary spermatogonia have many differences in cell size, organdie types and quantities as well as the distribution of regions in the testis. Chromatoid body was presented in most of germ cells, except for sperm. Nuclear bubbles were formed in the growth of sperm cells, and disappeared with the sperm maturation. The spermatozoon lacks an acrosome in its head which mainly occupied by nuclear. The tail mainly composed by the axoneme which present a typical "9 +2" structure.3. Study on the artificial propagation of 5. chongiSperm’s fast movement time and life time amounting to 43±8 s and 180±36 s separately in natural river water; The duration in which sperm kept fast motility and the sperm life span was basically similar in NaCl and glucose solutions; Sperm acquisition of high motility in 0.6% NaCl solution and 0.4% glucose solution, the average fast motility time was 54 s and 52 s, and the average life time was 1508 s and 1760 s separately; The sperm which lost their motility would reactivate motility if joined the water.During the year 2005～2006, in order to study the technology of artificial propagation, four kind of different combination hormones were injected separately into 180 mature wild S.chongi, which have been artificial domesticated in running water fishpond. Altogether, the quantity of fertilized eggs laid by breeding fish was 500,000 and the 90,000 larva were hatched out. The water temperature scope which artificial propagate can be successfully implemented was form 13℃to 18℃. The optimum combination of exterior hormones was: the combination of PG and HCG, with two injections, the first time injection dosage was (5mg+600IU)/kg fish body weight, and the second time was (12mg+1000IU)/kg fish body weight, the reactive time was about 85~98 h under the water temperature 13～18℃.Under the water temperature 17±1℃, the embryonic development process took 124 hours form fertilization to hatching with the average cumulative temperature of 2108.00 h·℃. The yolk-sac larvae started to swim horizontally with inflation the pneumatic cyst 9 days after hatching. The embryonic development process could be divided into 6 stages, including fertilized egg stage, cell division stage, blastula stage, gastrula stage, neural stage and organ formation stage, and contained twenty-four developing phrases as other teleost. each of their characteristic was described. The developmental speed of embryo and yolk-sac larva of S.chongi was quicker in the Schizothoracinae, later would be specially more obvious in the muscle effective phase; The frequency of palpitation was not simple linear growth from slowly to the quickly, but was a curve change process with speed interaction; each stage of early development has different sensitivity to (peripheral environment, the gastrula stage was more sensitive than other stages.更多还原