【作者】 崔淼；

【导师】 赵俊； 陈湘粦；

【作者基本信息】 华南师范大学 ， 动物学， 2003， 硕士

【摘要】 采用聚丙烯酰胺凝胶垂直板电泳技术，对同源和异源精子激发彭泽鲫雌核发育胚胎发育过程和子代成体不同组织中的同工酶的表达情况进行了比较研究。结果表明同源和异源精子激发的胚胎发育过程在MDH、LDH和SOD同工酶表达上存在明显差异，充分说明了“异精生物学效应”的存在。MDH的差异有两点，①在同源精子激发的胚胎中，MDH6’表达至心博期后消失，但在异源精子激发的胚胎中，MDH6’持续表达至孵出期；②在孵出期，异源精子激发的胚胎比同源精子激发的胚胎多出两条带MDH4’和MDH5’。LDH的差异表现在异源精子激发的胚胎在原肠中期到肌肉收缩期比同源精子多出4条带(LDH9’～12’)。SOD同工酶差异表现在异源精子激发的胚胎在眼黑色素期和孵出期比同源精子激发的胚胎多出两条酶带(SOD1’、SOD2’)。而在EST和ME同工酶表达上，同源和异源精子激发的胚胎则几乎没有差异。对不同精子激发彭泽鲫雌核发育子代的生长效应研究，证实了异精效应的存在，发现海鲤精子是一种激发效果较好的外源精子。 对异精激发彭泽鲫雌核发育子代及其母本彭泽鲫和父本海鲤的血清、肌肉蛋白以及心、肝、肾、脑、眼等组织的酯酶(EST)、苹果酸脱氢酶(MDH)、苹果酸酶(ME)、超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)电泳图谱的比较研究。结果发现：异源子代的血清、肌肉蛋白和同工酶的电泳图谱与母本彭泽鲫相同而与父本海鲤显著不同，推测异源精子的作用可能是在DNA水平上对后代产生影响，这种影响没有表现在成体同工酶的表达上。 通过对三种鲫鱼品系不同组织同工酶的电泳图谱的比较研究，揭示了彭泽鲫和银鲫至少在生化水平上已有明显的分化，很可能起源于不同的地区，独立演化而形成。彭泽鲫和银鲫D系的同工酶电泳图谱都包括有野鲫的基本酶带，而彭泽鲫和野鲫的酯酶同工酶电泳图谱尤为相似，推测彭泽鲫和银鲫都起源于野鲫，彭泽鲫和野鲫的亲缘关系较近，而银鲫和野鲫的亲缘关系较远。更多还原

【Abstract】 GUI Miao (College of Life Science, South China Normal University, Guangzhou 510631)Pengze crucian carp, Carassius auratus of Pengze, is a unique model system for studying evolutionary genetic and selective breeding, owing to its specific genetic background and reproductive modes. The species exists a triploid bisexual population, female outnumbering male by about four to one in their natural habits, and reproduces by gynogenesis. Surprisingly, heterogeneous sperm from other species of fish, besides stimulating and activating development of the egg, are able to contribute to the phenotype of gynogenetic offspring, that is allgynogenesis.The patterns of expressions in five isozymes (EST, MDH, ME, SOD and LDH) were investigated in the embryogenetic stages of gynogenetic crucian carp(Carassius auratus of Pengze) activated by homogenous and heterogeneous sperms. Five isozymes (EST, MDH, ME, SOD and LDH) of different tissues, serum and muscle protein of Bumper crucian carp (C. auratus of Pengze (3-)XCyprinus acutidorsalis ( & )), its female parent crucian carp (C. auratus of Pengze) and male parent seap carp (C. acutidorsalis) were also analyzed with vertical polyacrylamide gel electrophoresis.The patters of MDH, LDH and SOD showed apparent differences between the embryo activated by homogenous sperm and that activated by heterogeneous sperm. The main differences in MDH exist in hatching stage, which showed two more bands(MDH4’ and MDH5’) in the isozyme pattern of embryo activated by heterogeneous sperm than that activated by homogenous sperm. The main differences in LDH exist in five stages from middle-gastrula to muscle construction, which showed four more bands (LDH9’, LDH10’, LDH 11’ and LDH 12’) in the isozyme pattern of embryo activated by heterogeneous sperm than that activated by homogenous sperm. The main differences in SOD exist in two stages-6-from darkly-pigmented eye to hatching, which showed two more bands(SODl’ and SOD2’) in the isozyme pattern of embryo activated by heterogeneous sperm than that activated by homogenous sperm. Meanwhile, the other two isozymes (EST and ME) detected in this paper did not show apparent differences between the embryo activated by homogenous sperm and that activated by heterogeneous sperm. Furthermore, the speed of development of the embryo activated by heterogeneous sperm is slight faster than that activated by homogenous sperm. By comparing the growth of the offspring of Pengze crucian carp produced by various sperm stimulating, the results comfirmed that the growth effects exist in the offspring activated by heterogeneous sperm, and discovered that seap carp can finely activated the gynogenesis of Pengze crucian carp.The results showed that the electrophoretogram patterns of isozymes, serum and muscle protein of Bumper crucian carp were the same as those of its female parent and significantly different from its male parent. It is speculated that the function of heterogeneous sperm was only in DNA level and not behave on the isozyme expression of grown progeny.Isozymes in different tissues of three stains of crucian carp were studied comparatively. The results indicated that Pengze crucian carp is different from Silver crucian carp in biochemical level, they probably came from different region. The electrophoretogram of Pengze crucian carp and Silver crucian carp all included the basal patterns of Wild crucian carp. EST isozyme patterns between Pengze crucian carp and Wild crucian carp is similar especially among the patterns of several isozyme. It is speculated that Pengze crucian carp and Silver crucian carp were all origined from Wild crucian carp. The relations between Pengze crucian carp and Wild crucian carp is closer than that of Silver crucian carp and Wild crucian carp.更多还原