尼罗罗非鱼六个性别相关标记的FISH分析

从尼罗罗非鱼（Oreochromis niloticus）BAC基因文库5个克隆中提取和纯化含有6个性别连锁或相关标记（CLC5，GM204，GM271，GM354，UNH995和UNH104）的重组质粒DNA作为模板，以简并核苷酸为引物，通过PCR制备原位杂交探针。探针用荧光素进行标记，并与尼罗罗非鱼中期相染色体进行荧光原位杂交以确定这些标记在尼罗罗非鱼染色体上的位置和分布。结果显示，这些性别连锁或相关标记都位于尼罗罗非鱼第一对染色体长臂近末端，从分子细胞学角度验证了第一对染色体是尼罗罗非鱼的性染色体。另外由于这些标记的荧光信号在XY个体的2条性染色体上都有，一方面说明这些标记在罗非鱼上还不是性别特异的；另一方面也验证了尼罗罗非鱼的性染色体还处于分化的早期阶段。【中国水产科学，2006，13（4）：525—529】     

Reproductive performance,survival and development of nauplii and copepodites,sex ratio and adult life expectancy of the harpacticoid copepod,Euterpina acutifrons,fed different microalgal diets

A series of laboratory experiments were conducted on the harpacticoid copepod, Euterpina acutifrons, to assess the influence of 10 different microalgal diets (four monoalgal and six mixed algal diets) on several parameters related to its productivity in culture. The four monoalgal diets were the Tahitian strain of Isochrysis sp. (T‐Iso), Pavlova salina (Pav), Tetraselmis chuii (Tet) and the diatom Chaetoceros muelleri (Chaet), the five binary diets were T‐Iso+Tet, Pav+Tet, T‐Iso+Pav, Tet+Chaet and Pav+Chaet, while the tri‐algal diet was T‐Iso+Tet+Pav. All diets were fed to copepods at 1500 μ gC L^?1 and in the case of binary or trialgal diets, carbon concentration was divided equally between the two or three algae offered. Among monoalgal diets, the diatom Chaet was excellent for E. acutifrons. Out of the 10 diets tested, the binary diet Tet+Chaet, which contained the diatom Chaet, was the best for naupliar production of single pair E. acutifrons (19.5±1.7 nauplii female^?1 day^?1), significantly (P<0.05) higher than all other treatments except for the Chaet treatment (P>0.05). Similarly, in the group naupliar production trial (50 adult E. acutifrons per replicate), Tet+Chaet produced a significantly higher number of nauplii (405.8±18.6 nauplii day^?1) than the other treatments (P<0.05). Tet+Chaet further supported the highest naupliar survival (82.0±2.8%) and copepodite survival (89.0±2.8%), while the mono‐algal diet Chaet produced the second highest naupliar (76.7±2.6%) and copepodite survival (83.5±2.6%). In contrast, Pav produced the lowest overall survival at the naupliar stage (30.0±2.9%), significantly lower than all other treatments (P<0.05). While development from newly hatched nauplii to copepodites was not significantly affected by diets, mean development time from nauplius to adult was significantly different among treatments. Mean development time from hatching (naupliar I stage; NI) to the adult stage was the fastest with Tet+Chaet and Chaet (6.8±0.0 days for both treatments), which was significantly faster than that of Pav, T‐Iso Pav+Tet and T‐Iso+Pav+Te treatment (P<0.05). E. acutifrons sex ratio was significantly affected by diets, and always skewed in favour of males. Feeding on Pav resulted in the lowest proportion of females (23.7±1.2%), significantly lower than for six of the other treatments (P<0.05). Adult females had longer average life expectancy than males for all treatments, and were the longest when fed Tet+Chaet (9.5±0.4 days), which was more than twice as long as the shortest lifespan recorded for the Pav treatment (4.2±0.6 days) (P<0.05). In summary, among 10 diets tested, the binary diet Tet+Chaet appeared to support the highest culture productivity of E. acutifrons while the diatom Chaet also performed well as a monoalgal diet.     

全雄奥利亚罗非鱼的制种与应用研究

为获得全雄奥利亚罗非鱼(Oreochromis aurea),对WZ♀-ZZ♂类型的奥利亚罗非鱼采用原系(ZZ♂)与转化系(ZZ△♀)二系配套技术,控制遗传全雄鱼的性别。根据泄殖器特征对转化后的实验鱼进行性別鉴定,确定性别。对实验鱼的外部形态、内部结构进行生物学描述。统计并观察实验鱼的雄性率、生长和发育状况。总结全雄奥利亚罗非鱼在应用中的优势,以期为奥利亚罗非鱼养殖业的发展指明方向。     

Morphological sex change upon treatment by endocrine modulators in meiogynogenetic tench (Tinca tinca L.)

Exogenous steroids alter sex differentiation in fish substantially. In the present study we have evaluated the effects of 17α-methyltestosterone (MT) and oestrogen receptor antagonist Tamoxifen (TA) on gonadal development and skewness of the sex ratio in all-female tench juveniles. In the first two experiments, sexually undifferentiated juveniles were orally treated with three doses of MT and TA (50, 100 and 150 mg kg⁻¹). Both the treatments resulted in a moderate dose-dependent masculinization, with neomale production ranging from 17% (50 mg kg⁻¹) to 26% (150 mg kg⁻¹) for the MT only treatment, and from 0% (50 mg kg⁻¹) to 27% (100 mg kg⁻¹) only for the TA treatment respectively. In the third experiment treatment of sexually differentiated tench females with single steroid treatments or combinations of the two resulted in populations composed of females and intersex individuals. The significantly highest occurrence of intersex individuals (45.5%) was found in the group subjected to combine treatment of MT+TA (150+200 mg kg⁻¹). No masculinization effect of the single or the combined treatment occurred. It can be concluded that oral treatment with MT or TA only slightly modifies the normal process of sex differentiation in gynogenetic tench juvenile, but treatment with the above-mentioned combinations has a highly significant potential to skew the sex ratio in sexually differentiated tench females. However, from an applied point of view, the treatment procedure will need optimization before use at a commercial level.     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

A genetic tool for evaluating male‐mediated stock introgression in Atlantic salmon

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养殖银鲳性腺发育规律和性类固醇激素变化

为掌握养殖银鲳的性腺发育规律及其与性类固醇激素水平的关系,本研究采用组织切片技术和酶联免疫吸附法(Elisa),观察养殖条件下银鲳的精巢与卵巢发育特征和血清中性类固醇激素的周年变化,并分析性腺成熟指数(GSI)、肝重指数(HSI)和肥满度(CF)与性腺发育的关系。结果显示,养殖银鲳雌鱼1龄即可发育成熟,成熟卵巢呈一对""形的囊状结构。11月份有近一半个体发育至III期,33.3%已发育至IV期,12月—翌年1月越冬期间约2/3退化至II期,3月又迅速发育至IV期,4月份达到V期并产卵,5月份产卵结束卵巢进入恢复期IV期。银鲳精巢属小叶型,III期精巢出现小叶腔,IV期精巢出现精子,精巢发育速率快于卵巢,即"雄性早熟",7月龄约20%个体可达到性成熟,越冬时退化至III期,3月重新发育,精子分批成熟排精。雌鱼GSI值为0.19%~12.89%,HSI为0.97%~2.30%,CF为2.30~3.08 g/cm~3,雄鱼的GSI为0.08%~2.62%,HSI值为0.73%~1.83%,CF为2.11~2.80 g/cm~3,GSI值在V期时达到最大,HSI和CF值则于IV期达到峰值。雌鱼血清中雌二醇(E2)周年表达水平为23.27~59.13 pg/m L,雄鱼为15.90~36.20 pg/m L,雌鱼的睾酮(T)为14.57~68.67 nmol/L,雄鱼的T为18.62~66.49 nmol/L,E2水平与HSI值呈现显著正相关关系,T表达水平与GSI值呈现显著正相关关系。研究表明,银鲳血清中性类固醇激素含量与精巢和卵巢的发育密切相关,可作为了解养殖银鲳性腺发育的重要指标。     

Sex change in coral reef fish

Gonadal differentiation can take many forms in fish, ranging from gonochorism, where individuals directly develop as male or female and finally possess only testis or ovaries at sexual maturation, to hermaphroditism where the same individuals can produce mature male and female gametes at some time in their lives. Hermaphrodite fish are, thus, an excellent model for studying the plasticity of sex determination and differentiation in vertebrates. We have shown that sex steroids play a principal role in sex differentiation and sex change in fish. Our laboratory implements several fish models that undergo sex change from female to male or male to female or in both directions. In this review, we will briefly discuss recent advances in our understanding of the mechanism of sex change in coral reef fish.