Zhu L F, Gui J F
State Key Laboratory for Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072.
Shi Yan Sheng Wu Xue Bao. 1998 Dec;31(4):369-76.
Isozyme zymograms of esterase (EST), lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and superoxide dismutase (SOD) were analysed by polyacrylamide gradient gel electrophoresis at different developmental stages of embryogenesis in 4 types of various ploidy crucian carp embryos, including haploids, diploids, natural triploids, and multiple tetraploids, and 2 types of haploid and diploid common carp embryos. Haploid embryos of crucian carp (Carassius auratus) and common carp (Cyprinus carpio) were produced by treating eggs with UV-irradiated milt from blunt snout bream (Megalobrama amblycephala). Natural triploid embryos were obtained from the eggs of gynogenetic silver crucian carp (Carassius auratus gibelio) inseminated with milt from red common carp. Multiple tetraploid embryos were also produced by gynogenesis from eggs of the newly discovered multiple tetraploid females inseminated with milt from red common carp. Gradient gel electrophoresis indicated that the band types and staining intensity of 4 isozymes expressed in haploid embryos of crucian carp and red common carp were similar to that in the correlative diploid embryos. In natural triploid silver crucian carp embryos, the zymograms of MDH and SOD isozymes were identical with that of diploid crucian carp embryos, but the EST and LDH isozymes manifested more new enzyme bands in comparison with diploid embryos. The corresponding expressed products of some bands in the triploid embryos, such as EST5 and EST6, could be observed also in red common carp embryos, which provided evidence for hybrid origin about the gynogenetic fish. The multiple tetraploids incorporated one foreign genome of red common carp, therefore, the effects of genes from the foreign genome could be observed in the multiple tetraploid embryos. Gene expression of the isozymes in the tetraploid embryos was somewhat similar to that in hybrids. Owing to interaction of triploid silver crucian carp genomes and common carp haploid genome, some isozyme bands, such as EST5 and EST6, changed in quantity, and some bands increased, such as s-SOD1, s-SOD2, s-SOD3 and s-SOD4 in the tetraploid embryos. Moreover, the heterogeneity was revealed among embryos developed from gynogenetic eggs of 3 different multiple tetraploid individuals.
采用聚丙烯酰胺梯度凝胶电泳法,对4种不同倍性鲫鱼胚胎(包括单倍体、二倍体、天然三倍体和复合四倍体)以及2种单倍体和二倍体鲤鱼胚胎在胚胎发育不同阶段的酯酶(EST)、乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)和超氧化物歧化酶(SOD)同工酶酶谱进行了分析。鲫鱼(Carassius auratus)和鲤鱼(Cyprinus carpio)的单倍体胚胎是通过用经紫外线照射的团头鲂(Megalobrama amblycephala)精液处理鱼卵产生的。天然三倍体胚胎是由雌核发育的银鲫(Carassius auratus gibelio)鱼卵与红鲤精液授精获得的。复合四倍体胚胎也是通过用新发现的复合四倍体雌鱼的鱼卵与红鲤精液进行雌核发育产生的。梯度凝胶电泳表明,鲫鱼和红鲤单倍体胚胎中4种同工酶的条带类型和染色强度与相关二倍体胚胎相似。在天然三倍体银鲫胚胎中,MDH和SOD同工酶的酶谱与二倍体鲫鱼胚胎相同,但EST和LDH同工酶与二倍体胚胎相比表现出更多新的酶带。三倍体胚胎中某些条带(如EST5和EST6)的相应表达产物在红鲤胚胎中也能观察到,这为雌核发育鱼类的杂交起源提供了证据。复合四倍体整合了红鲤的一个外源基因组,因此,在复合四倍体胚胎中可以观察到外源基因组基因的作用。四倍体胚胎中同工酶的基因表达与杂种中的有些相似。由于三倍体银鲫基因组与鲤鱼单倍体基因组的相互作用,四倍体胚胎中的一些同工酶条带(如EST5和EST6)数量发生变化,一些条带增加,如s-SOD1、s-SOD2、s-SOD3和s-SOD4。此外,在3个不同复合四倍体个体的雌核发育鱼卵发育的胚胎之间显示出异质性。