Effect of microtubule reactive drugs on steroid- and centrifugation-induced germinal vesicle migration during goldfish oocyte meiosis

During the process of progestogen-induced meiotic maturation in the goldfish oocyte, the oocyte nucleus (germinal vesicle, GV) migrates to the sperm entry site or micropyle at the animal pole. Following GV migration (GVM) to the micropyle, the nuclear membrane undergoes dissolution (GVD) and the cell enters metaphase I in preparation to generate the first polar body. Microtubule destabilizing drugs including colcemid, nocodazole and vinblastine were found to elicit GVM, mimicking the process which occurs just prior to the prophase I-metaphase I transition during steroid induced oocyte meiotic maturation. In addition, these drugs enhanced the induction of GVM by 17 alpha, 20 beta dihydroxy-4-pregnen-3-one, a potent, naturally occurring meiotogenic steroid in this species. By contrast, taxol, a microtubule stabilizing drug, was found to inhibit steroid induced GVM. A new assay for centrifugation induced GVM was applied to the goldfish oocyte in order to assess effects of steroids and drugs on GVM, without the complication of GVD or the restrictions imposed by the slow time course of naturally occurring GVM. The effective centrifugal force (ECF) required to elicit GVM in 50% of the oocytes (ECF50) decreased significantly after short incubations (1-5 hr) of oocytes with either 17 alpha,20 beta dihydroxy-4-pregnen-3-one or microtubule disrupting drugs (i.e., colcemid, nocodazole, or vinblastine). A working hypothesis, modeled after the effects of microtubule disrupting agents on intermediate filament arrays in somatic cells, is proposed in which a small number of microtubules or other polymeric tubulin units are responsible for maintaining a cytoskeletal array.(ABSTRACT TRUNCATED AT 250 WORDS)     

滇池高背鲫和方正银鲫酯酶、乳酸脱氢酶同工酶的比较研究

本研究以方正银鲫(Carassius auratus gibelio Bloch)普通鲫(Carassius auratus)和滇池高背鲫(Carassius sp.)的各种组织器官为材料,进行酯酶(Esterase)和乳酸脱氢酶(LDH)同工酶电泳图谱的分析比较。结果表明:9种不同组织中酯酶同工酶谱带各不相同,有明显的组织特异性。滇池高背鲫的酯酶谱图有3种表型。方正银鲫和滇池高背鲫同一组织的LDH同工酶酶谱也有明显差异。等电聚焦凝胶电泳(T=7.5％,C=5％)的结果又表明这二种鱼的肝脏、脑、卵的酯酶同工酶酶谱及电泳扫描图亦有差异。这些结果揭示滇池高背鲫与方正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,由不同的祖先,独立演化而形成。滇池高背鲫与云南普通鲫的LDH酶谱较为接近,这说明滇池高背鲫最可能起源于云南本地的普通鲫。     

鲫鱼后鳃体结构与功能的研究

本文对鲫鱼后鳃体的形态结构及其功能进行了初步研究。结果表明,鲫鱼后鳃体是一种内分泌腺体。腺体由滤泡组成,滤泡上皮包括二种颗粒细胞(亮细胞和暗细胞)和一种支持细胞。对不同性腺期的鲫鱼后鳃体的观察表明,后鳃体滤泡细胞随性腺发育增加高度,滤泡增生,腺体体积增大。而性腺成熟后期,后鳃体滤泡上皮有崩溃现象。揭示后鳃体可能与性腺发育有关。用免疫组织化学的非标记过氧化物酶技术对鲫鱼后鳃体进行激素降钙素的定性定位研究,观察到滤泡上皮的颗粒细胞发生阳性反应,由此证明颗粒细胞可分泌降钙素。     

~(90)锶对鲫鱼胚胎发育的影响及鲫仔鱼对~(90)锶吸收积累的研究

在胚胎发育过程中,用不同比放射性的~(90)锶试液处理受精卵,各处理组都有畸形和死亡现象。畸形的高峰一般出现在囊胚期和晶体形成期,畸形的类型一般为脊椎弯曲、体短、胸腔和卵黄囊扩大,头和眼畸形。随后即出现死亡高峰。例如:游动期的死亡率分别是:21%(对照);36%(5×10~(-11)居里/升);40%(5×1-~(-10)居里/升);45%(5×10~(-9)居里/升)。鲫仔鱼对~(90)锶吸收积累的结果表明:鲫仔鱼体中比放射性大小与试液的比放射性大小成正比,并随试验吋间的延长,生物量的增加而增加,鲫仔鱼的积累系数则不依试液的比放射性大小和生物量的多少为转移。     

水体pH胁迫对异育银鲫皮质醇激素和非特异性免疫的影响

实验比较了不同pH胁迫对异育银鲫(Carassius auratus gibelio)皮质醇激素和非特异性免疫的影响。3个实验组水质的pH分别为6.0、8.5、9.5,对照组是pH为7.4的曝气自来水。分别在实验的第0、14和30天采血,血浆用于测定皮质醇含量和溶菌酶活性;分离的白细胞,采用流式细胞术测定吞噬活性和呼吸暴发。在实验的第4天多取1次血样,用于皮质醇含量测定。在实验的第31天,测量鱼的体重,并用柱状黄杆菌(Flavobacterium columnare)浸泡攻毒,7d后统计死亡情况。结果显示:在pH 6.0的胁迫下,白细胞的吞噬活性在第14、30天显著降低,而呼吸暴发和皮质醇含量没有显著变化,溶菌酶活性和生长与对照组没有显著差异,用黄杆菌攻毒也没有出现死亡。而在pH 8.5和9.5的胁迫下,皮质醇含量在第14、30天显著升高,白细胞的吞噬活性、呼吸暴发显著降低;在pH 9.5胁迫下的溶菌酶活性显著降低,用柱状黄杆菌攻毒两组都出现死亡。实验表明pH为8.5和9.5的胁迫严重影响了异育银鲫的非特异性免疫功能,流式细胞术能较客观测定白细胞的吞噬活性和呼吸暴发,可用于评价胁迫对鱼类健康的影响。     

鲫鱼微卫星DNA的分离及多态性分析

目的：用近缘物种鲤微卫星引物来分离鲫鱼微卫星标记并对其多态性进行分析。方法：以鲫鱼基因组DNA为模板,采用6对鲤微卫星引物进行PCR扩增,PCR产物经8%的非变性聚丙烯酰胺凝胶电泳和银染色检测。结果：筛选出2个以AC和TA为重复单元的鲫鱼新的微卫星标记。多态性分析表明,这2个微卫星标记的遗传杂合度分别为0.611和0.644,多态信息含量为0.536和0.572,属于高度多态性标记。结论：该研究筛选的2个微卫星标记可应用于鲫鱼遗传多样性、遗传连锁图谱构建及分子标记辅助育种等方面的研究。     

彩鲫hira基因片段的克隆及表达分析

hir／hira基因最先作为组蛋白基因表达的一种负调节因子从酵母中被鉴定出来。现已证实,HIRA包含一组保守的蛋白家族,广泛存在于低等真核生物、无脊椎动物和脊椎动物等多种生物体当中,为生命发育所必需。关于hira基因在动物发育过程中的具体作用研究还不是很多,为了进一步探讨hira基因在鱼类发育过程中的作用,作者根据已知hira基因保守序列设计一对简并引物,分别以彩鲫基因组DNA和卵巢cDNA为模板克隆了彩鲫hira基因（Cahira）片段,该片段基因组序列为2181bp,基因组片段含有6个内含子,长度分别为118bp、275bp、372bp、84bp、472bp、86bp;cDNA序列为759bp,编码253个氨基酸,具有5个WD结构域。对其氨基酸序列进行比较,结果表明,彩鲫hira基因的氨基酸序列与河鲍、爪蟾、鸡、小鼠、人的hira基因氨基酸序列具有非常高的同源性,分别高达92％、89％、89％、87％和88％。组织特异性表达分析表明,所检测的彩鲫组织除了精巢和肌肉中未检测到hira基因表达外,其余组织均有表达。其中卵巢和肝脏中表达很强,而在脑、心、脾、肾中表达较弱,说明该基因可能在维持卵巢和肝脏组织的功能方面起一定作用。     

Isolation and characterization of polymorphic trinucleotide microsatellites of the polyploid crucian carp (Carassius auratus)

In this study, we isolated and characterized nine polymorphic trinucleotide microsatellites (CAG and CCT) from the crucian carp (Carassius auratus). The number of alleles per locus ranged from two to six. Five loci showed a significantly excess homozygosity, and a genetic linkage between CAL0102 and CAL0495 was strongly suggested. Our results confirmed the triploidy of Korean individuals, and the microsatellites were found to be useful for analysing the allelic state of the polyploid crucian carp.     

Extensive hybridization reveals multiple coloration genes underlying a complex plumage phenotype

Coloration is an important target of both natural and sexual selection. Discovering the genetic basis of colour differences can help us to understand how this visually striking phenotype evolves. Hybridizing taxa with both clear colour differences and shallow genomic divergences are unusually tractable for associating coloration phenotypes with their causal genotypes. Here, we leverage the extensive admixture between two common North American woodpeckers—yellow-shafted and red-shafted flickers—to identify the genomic bases of six distinct plumage patches involving both melanin and carotenoid pigments. Comparisons between flickers across approximately 7.25 million genome-wide SNPs show that these two forms differ at only a small proportion of the genome (mean F_ST = 0.008). Within the few highly differentiated genomic regions, we identify 368 SNPs significantly associated with four of the six plumage patches. These SNPs are linked to multiple genes known to be involved in melanin and carotenoid pigmentation. For example, a gene (CYP2J19) known to cause yellow to red colour transitions in other birds is strongly associated with the yellow versus red differences in the wing and tail feathers of these flickers. Additionally, our analyses suggest novel links between known melanin genes and carotenoid coloration. Our finding of patch-specific control of plumage coloration adds to the growing body of literature suggesting colour diversity in animals could be created through selection acting on novel combinations of coloration genes.