三个三倍体鲫鱼品系及野鲫mtDNA的比较研究

用16种限制性内切酶研究了银鲫(3N=156～162)、彭泽鲫(3N=162)和缩骨鲫(3N=150)3个三倍体鲫鱼品系及野鲫(2N=100)的线粒体DNA。有6种酶在种系间和种系内产生限制性片段长度多态性(RFLPs),银鲫共存在4种单倍型,彭泽鲫2种,野鲫3种,缩骨鲫1种。彭泽鲫和银鲫拥有相同的常见单倍型,缩骨鲫的单倍型属于野鲫的常见型。根据限制性位点的变异数据,计算了单倍型间的相似性、核苷酸多样性、品系内核苷酸多样性和品系间的遗传距离,确定彭泽鲫属于银鲫的一个地方品系,缩骨鲫属于野鲫的一个地方品系。根据核苷酸的差异,推算出银鲫和野鲫两个亚种的分化大约在11万年前完成。     

PCR survey of hox genes in the goldfish Carassius auratus auratus

A tetraploidization event took place in the cyprinid lineage leading to goldfishes about 15 million years ago. A PCR survey for Hox genes in the goldfish Carassius auratus auratus (Actinopterygii: Cyprinidae) was performed to assess the consequences of this genome duplication. Not surprisingly, the genomic organization of the Hox gene clusters of goldfish is similar to that of the closely related zebrafish (Danio rerio). However, the goldfish exhibits a much larger number of recent pseudogenes, which are characterized by indels. These findings are consistent with the hypothesis that dosage effects cause selection pressure to rapidly silence crucial developmental regulators after a tetraploidization event.     

A BAC library for the goldfish Carassius auratus auratus (Cyprinidae, Cypriniformes)

A goldfish (Carassius auratus auratus) bacterial artificial chromosome genomic library (BAC library) was constructed from one aquarium-bred male specimen (tetraploid, 4n=100, genome size=3.52 pg/cell). The library consists of 128,352 positive clones with an average insert size of 150.4 kb, covering the genome 11-fold. All clones were spotted onto nylon filters and thus are available for screening of genomic regions of interest, such as candidate genes, gene families, or large-sized syntenic DNA regions of cyprinid species. Preliminary screens with two genes were conducted with hybridizing probes to the genes RAG1 and lgi1. RAG1 is a single-copy gene in zebrafish and is duplicated in C. a. auratus. We found a very close correlation between the number of positive BAC clones and the expected library coverage. Two copies of lgi1 were found in zebrafish. We have detected four different copies in C. a. auratus, not in the expected abundance, which indicates some variation in the coverage of the BAC library. The preliminary screens indicate that many duplicated genes that resulted from the ancient fish-specific genome duplication persist in the tetraploid goldfish genome. Hence, the BAC library will provide a useful resource for the future work on comparative genomics, polyploidy, diploidization, and evolutionary genomics in fishes.     

负压状态下压力变化导致鲫鱼身体组织的损伤

通过试验研究负压状态下压力变化过程对鲫（Carassius auratus auratus)的损伤,试验采用真空泵和空气压缩机在试验容器内形成不同的压力变化过程,统计不同体长的鲫经历压力变化过程后的损伤情况,并对部分受损伤的鲫进行解剖和组织观察。研究发现负压状态下压力时变导数较大的变化过程会对鲫的生存构成直接威胁,主要损伤是鱼鳔部分或全部受损,在肝胰脏、肾脏等处有明显出血点。综合分析不同试验条件对鲫损伤的情况,得到了对鲫尽可能安全的压力时变导数极限值,从而为新型环保水力设施的设计提供参考依据,起到保护渔业资源的作用。     

Characterization of microsatellites located within the genes of goldfish (Carassius auratus auratus)

The goldfish (Carassius auratus auratus; Cyprinidae) is not only an important ornamental fish species, but also a useful model for biological studies. The sequence of goldfish genes present in the public database was searched for short tandem repeats, and 11 polymorphic microsatellites were detected within eight important genes. Two microsatellites were located in coding regions of the c‐myc and GAP‐43 genes, respectively, whereas the others were mainly located in 5′ and 3′ untranscribed regions of other genes, such as gonadotropin and activin. The average allele number of these microsatellites was 5.5 per locus with a range between 3 and 9. These microsatellites will be useful for ecological and population genetic studies of this species, as well as for the ornamental fish industry.     

金鱼ABC转运蛋白基因家族成员鉴定及分析

腺苷三磷酸结合盒转运蛋白(ATP-binding cassette transporter,ABC transporter)基因家族在原核生物和真核生物中广泛存在,该家族蛋白能够利用ATP裂解产生的能量将多种底物转运到膜上,参与多种生物过程,如营养摄入、细胞解毒、脂质稳态、信号转导、病毒防御以及抗原呈递等。目前,鱼类中,只在斑马鱼、斑点叉尾鮰和鲤鱼等少数鱼类中对该基因家族进行了系统的研究,关于金鱼ABC转运蛋白基因家族的详细分析,未见报道。本研究中,我们利用三代结合二代测序技术构建的金鱼转录组参考基因集数据,鉴定出55个ABC转运蛋白基因,通过系统进化分析将它们分为8个亚家族(A^H)。即金鱼ABC转运蛋白基因是由10个ABCA、14个ABCB、13个ABCC、5个ABCD、1个ABCE、4个ABCF、7个ABCG和1个ABCH组成。同时,我们将金鱼与斑马鱼、斑点叉尾鮰和鲤鱼等物种ABC转运蛋白基因家族成员的数目进行比较分析,推测硬骨鱼类特异的第3次全基因复制(3R-WGD)和谱系特异的第4次全基因组复制(4R-WGD)对金鱼该基因家族成员数目的影响。本研究结果为金鱼ABC转运蛋白基因功能的研究提供了理论依据。     

Condition cycles in juvenile Pagrus auratus

Nutritional condition was measured in juvenile snapper Pagrus auratus (<200 mm fork length) using three indices: relative condition index (I_c), hepatosomatic index (I_H) and digesto-somatic index (I_D). In a laboratory starvation experiment, all three indices declined substantially over a 24-day period, but I_H was most sensitive. In wild snapper I_c and I_H showed no diel cycles. I_D for 0+ snapper showed a strong diel cycle consistent with continuous feeding during daylight hours and lack of feeding during the night. I_D for 1+ snapper showed no diel cycle. Subsequent analyses were restricted to daytime samples for I_H and morning samples for I_D to minimize the confounding effect of time of day. I_c, I_H and I_D were monitored at one site at approximately bi-monthly intervals over a period of 3.25 years. All three indices varied significantly, but only I_D and I_H displayed seasonal cycles. I_D peaked in late summer-autumn and dropped to a minimum in winter, due to seasonal fluctuations in the feeding rate that probably reflected variations in metabolic and growth rates. I_H peaked in autumn-winter and declined to a minimum in summer, thus lagging 4–6 months behind I_D. I_H varied significantly among four sampling sites for all five combinations of sampling periods and year classes, whereas I_c varied significantly among sites for only one of the five combinations. The Kawau Bay site, which supported the highest density of snapper, had the highest I_H for all except one of the combinations. This suggests that juvenile snapper aggregate selectively at sites that provide optimal feeding conditions. However, no relationship was found between I_H and growth rate, indicating that better nutritional condition may not translate into faster growth.     

鲫鱼雌性有关蛋白的研究

本文利用聚丙烯酰胺、SDS、超薄平板等电聚焦等多种电泳方法,对不同性别鲫鱼的血清分析后,发现了两种与雌性有关的特异血清蛋白,FSSP-Ⅰ及FSSP-Ⅱ。分离提纯了含量较多的FSSP-Ⅱ,其分子量、等电点、氨基酸组成分析的结果证明,它是一类与已报道过的鱼类雌性蛋白极不相同的、与性别有关的蛋白。为性别问题的研究提供了有意义的资料。     

Die Auswirkung der Übungsverteilung auf das Lernen und Vergessen bei Carassius auratus auratus Linné3

The significance of massed and distributed practice in discrimination learning to criterion by goldfish was studied. All animals were trained to discriminate between 6 horizontal black and white stripes (positive stimulus) and 2 vertical black and white stripes (negative stimulus). 5 groups of animals, 12 Ss each, got different training programs. Groups varied in number of trials per day (30, 10, 5 successive trials) and in distribution of trials over the day (30 trials given in blocks of 5 with ITI of 1 h or 5 trials, each spaced by an ITI of 1 h). In general animals with a small number of massed trials/day were superior in early acquisition period. When trials/day were spaced the method proved to be more efficient. The training method showed no effect concerning retention.     

不同品种金鱼和鲫鱼的分子系统发育关系研究

为探讨不同品种金鱼的系统进化关系,利用PCR技术扩增了金鱼的7个代表品种红龙睛(Red dragongoldfish)、红帽子(Red cap goldfish)、虎头(Tiger head goldfish)、琉金(Gold plating goldfish)、墨龙睛(Black dragongoldfish)、水泡眼(Water vesicle goldfish)、珍珠(Genuine pearl goldfish)的线粒体DNA上细胞色素b的部分核苷酸序列,长度为597 bp.结合GenBank中红鲫、野鲫、日本白鲫、银鲫、鲤鱼的序列进行比较分析,结果显示,这7个金鱼品种之间的同源性都很高,在99.5%～100%之间;7种金鱼和红鲫的同源性也很高,为99.5%～99.8%,与野鲫的同源性在96.8%～97.2%,与日本白鲫、银鲫的同源性为93.1%～94.3%,与鲤鱼的同源性相对较低,为88.3%～88.6%.利用DNAstar软件构建了不同品种金鱼和鲫鱼的分子系统树,从分子水平进一步证实了金鱼起源于野鲫.     

Early development of the chondrocranium in Chrysichthys auratus

The inception and development of the cartilaginous cephalis skeleton of Chrysichthys auratus is described from hatching to about 18 days post-hatching. At hatching, no skeletal structure is present. Not until day 3 do clearly delimited cranial primordia become apparent. As in many siluriforms, the neurocranium is platybasic from the start, the suspensorium constitutes, with Meckel's cartilage and the hyoid bar, a single cartilaginous element, and the junction between the front and rear of the neurocranium is complete on day 4. By day 8 the quadratomandibular joint has formed and the tectum posterius has appeared. Cartilage reduction first affects the trabecular bars, then, markedly, the visceral arches. By day 18 the braincase floor has almost disappeared.     

Gill sphaerosporosis in goldfish (Carassius auratus)

Infections caused by a Sphaerospora sp. resembling S. chinensis are reported for the first time in goldfish (Carassius auratus) from North America. The myxosporean was found in the respiratory epithelium of the gill of pond-reared fish. Spores from stained tissue sections were spherical with an equal mean length and width of 6.3 microns. Spore valves were thickened at the suture which lies in a plane perpendicular to two prominent pyriform polar capsules. The polar capsules were 4.0 x 2.8 microns in length and width. Both monosporous and disporous development within a surrounding "pseudoplasmodium" was detected. Infections caused moderate hyperplasia and occasional necrosis of the respiratory epithelial cells of the gill.     

Eosinophilic granule cells in Carassius auratus scale epidermis

Eosinophilic granule cells (EGCs) were characterized in Carassius auratus scale epidermis in situ and in explants. Live EGCs were readily identified by the presence of numerous large cytoplasmic granules observed with DIC microscopy. Histochemical staining with toluidine blue and alcian blue yielded granule metachromasia and pale blue granules, respectively, both consistent with mammalian mast cell staining. However, EGCs also share some features with mammalian basophils as neutral red dye was selectively incorporated into EGC granules. EGCs within scale epidermis were actively motile, displaying average speeds of 16 μm/min and maximum speeds of greater than 40 μm/min and showing morphological plasticity during migration. The predominant motile phenotype was elongate with a well‐developed leading lamella, while a broader body motile morphology was observed to a lesser extent. A trailing, relatively unchanged uropod was associated with every motile EGC and invariably contained one or a few granules. A rounded EGC shape without a leading‐edge or trailing uropod was also observed and was generally associated with static cells. Individual cells readily switched between the three major shapes during motility; static cells could abruptly develop a polarized morphology, and actively motile cells switched between elongate and broad‐bodied shapes or the static, rounded shape.     

Temperature tolerance in the goldfish,Carassius auratus

• 1.Lower and upper temperature tolerances of 240 goldfish, Carassius auratus, were measured at constant acclimation temperatures of 5, 15, 25 and 35 °C via critical thermal methodology.
• 2.Mean critical thermal minima and maxima ranged from 0.3 to12.6 °C and 30.8 to 43.6 ° C, respectively, and were significantly linearly related to acclimation temperature. Acclimation temperature accounted for approximately 90% of the variance in temperature tolerance. Ultimate critical thermal minimum and maximum equaled 0.3 and 43.6 °C, respectively.
• 3.Integrating the temperature tolerance polygon yielded an area of temperature tolerance of 1429 °C², which is approximately 17% larger than the polygon measured via the incipient lethal temperature approach. This difference is explained by methodological differences in these two techniques to quantify temperature tolerance.