人工诱导日本鳗鲡性腺发育组织学研究

从人工诱导日本鳗鲡(Anguill japonica)性腺成熟的角度,采用对亲鳗群体外形和解剖观察,以及性腺组织切片的方法,对日本鳗鲡从下海鳗到性腺发育成熟的全过程进行了组织学和细胞学研究。结果表明,人工诱导的日本鳗鲡精巢发育仍可分为5个时相,即精原细胞前增殖期(Ⅰ);精原细胞后增殖期(Ⅱ),精母细胞生长期(Ⅲ),精子出现期(Ⅳ)和精子成熟期(Ⅴ)。除日本鳗鲡下海鳗的精巢有73.3%的个体处在第Ⅰ时相外,其整个精巢的发育过程和细胞学特征与以前的研究无太大差异。在下海鳗群体中未观察到第Ⅰ时相的卵巢。因此,人工诱导日本鳗鲡的卵巢发育只有卵母细胞单层滤泡期(Ⅱ);脂肪泡出现期(Ⅲ);卵黄充满期(Ⅳ)和卵母细胞最后成熟期(Ⅴ)。本研究除对人工诱导下日本鳗鲡性腺发育各时相的组织学和细胞学特征进行了观察外,还探讨了人工诱导日本鳗鲡性腺发育的时间、下海鳗群体的组成、以及脂肪泡的形成和变化过程等问题,为以后的日本鳗鲡人工育苗技术提供了参考依据。     

胶州湾菲律宾蛤仔的性腺发育

１９９６年４～６月,对胶州湾内的菲律宾蛤仔连续取样,将性腺用１０％福尔马林液固定,组织切片４～６μｍ,Ｈ．Ｅ染色,显微照相显示菲律宾蛤仔卵巢和精巢的发育是同步进行的,性腺在４月初开始进入繁殖初期的快速发育,４月中和４月底生殖细胞已明显增大,至５月上旬染滤已被成熟的生殖细胞充满,５月中旬前后生殖细胞被分批排放,５月下旬和６月上旬性腺进入休止状态,至此,形成了一个繁殖期,在繁殖期内,高龄贝比低龄贝发育     

人工繁殖大鲵性腺组织学观察

对4尾人工繁殖的3龄成熟大鲵的性腺组织学进行观察。精巢中的生殖细胞分为5种,卵巢中的卵母细胞发育分为6个时相,并且在成熟的精巢和卵巢中观察到各个不同发育阶段的性腺细胞,说明大鲵性腺发育是不同步的,这对大鲵的种群繁衍具有积极的意义。     

稚鳖的性别分化及组织学研究

解剖出生１５～４５ｄ的稚鳖,所有个体的性腺呈细线状,肉眼分辨不出雌雄。组织学研究发现,雌雄个体的性别都已明显分化。雄性精巢的外壁由两层膜构成,其内部由许多隔膜分隔成许多精细管,精细管里充满原始的精母细胞;雌性卵巢有卵巢腔和输卵管,由结缔组织、毛细血管和生殖上皮形成卵巢膜和基质,并把卵巢分隔成一个个小室,原始的卵泡就在这些小室中生长、发育。     

人工养殖条件下云南盘鮈成鱼性腺发育及周年变化

采用常规解剖学和组织学方法,研究人工养殖条件下云南盘鮈（Discogobio yunnanensis）成鱼性腺发育及周年变化情况,为云南盘鮈全人工化繁殖提供基础资料。实验时间为2016年6月至2017年6月,每月中旬解剖成鱼取性腺,雌雄各5～8尾,活体状态下测量常规生物学数据,解剖后肉眼观察性别及性腺发育情况,并记录色泽等其他外形特征。取性腺称重并测量其长度和宽度,计算其性腺成熟系数（GSI）,观察其性腺发育状况。结果表明,云南盘鮈在人工养殖条件下可以性成熟,并能繁殖后代。卵巢发育可分为Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ期,Ⅱ期卵巢持续时间最长（6-11月）,11月开始出现Ⅲ期卵巢,12月主要为Ⅱ、Ⅲ期卵巢,次年1-3月主要为Ⅲ期卵巢,4-5月主要为Ⅳ期卵巢,5月底到达Ⅴ期即成熟期,排卵后,进入短暂的Ⅵ期,后迅速退化至Ⅱ期。本实验没有观察到Ⅰ期性腺。云南盘鮈精巢发育也可分为5期。6-12月主要为Ⅱ期精巢,12月开始出现Ⅲ期精巢,以Ⅱ、Ⅲ期精巢过冬;次年1-3月主要为Ⅲ期精巢,含少量Ⅱ、Ⅳ期精巢,4月主要为Ⅳ、Ⅴ期精巢,5月全为Ⅴ期精巢,6月则以Ⅱ期精巢为主,含少量Ⅴ、Ⅵ期精巢。云南盘鮈卵巢成熟系数为1%~18%,最大成熟系数18%;精巢成熟系数为0.2%~8.0%,最大成熟系数为8.0%;性腺成熟系数在4-6月出现一次高峰,成熟系数变化趋势与周年变化一致,说明云南盘鮈为一次产卵类型鱼类,且繁殖期集中在5月底至6月初。     

半滑舌鳎性腺的组织学研究

采用组织切片,对半滑舌鳎性腺的组织学结构进行了观察研究。结果表明,半滑舌鳎的精巢为小叶型,精巢内的生殖细胞分为五种。其精原细胞存在于精小叶的内壁上,精母细胞和成熟的精子细胞位于精小囊中。精子成熟后,精小囊破裂释放精子进入小叶腔,完成发育过程。半滑舌鳎卵巢外有被膜包裹,其被膜的结缔组织可明显地分为内外两层。内层的结缔组织、血管与原始生殖细胞一同伸进卵巢内部形成许多产卵板。产卵板以卵巢腔为中心向四周呈辐射状排列,上有原始生殖细胞分化形成卵原细胞和不同时相的卵母细胞。半滑舌鳎卵巢中的卵母细胞的发育可以分为5个时相。本文描述了各时相卵母细胞的形态学特征。并阐述了半滑舌鳎的产卵类型。     

黑龙江银鲫雌雄同体的组织学

从方正银鲫自交繁殖的当年鱼群体中发现约有２％左右的雌雄同体。外形观察两侧性腺的分配和着生方式十分复杂和多样。一般为一侧精巢,另一侧为卵巢,但常见到一侧性腺,精巢和卵巢交替着生,有时则从卵巢旁生出一个球状精巢或囊状组织,但未见精巢旁长出球状卵巢的。组织学观察,两种组织的界限分明,雌雄同体的精巢和卵巢都发育良好。精巢壶腹中充满各时相的精原细胞、精母细胞和精子。卵巢则充满Ⅳ期中的卵母细胞,但与精巢交接处的卵细胞或进入精巢中少量卵细胞则发育迟缓,一般处于Ⅱ、Ⅲ时相。非常奇特的是：从卵巢旁生长的小囊状组织,一半为精巢,一半为卵巢,精巢发育良好,可见到各时相的精细胞,但卵巢发育不良,大多是Ⅱ、Ⅲ时相的卵细胞。两种组织交接处有膜分隔,同时有血管通过,供两组织营养。     

脊尾白虾的性腺发育及组织结构观察

为系统研究脊尾白虾的性腺发育及组织学特征,采用常规的石蜡切片及H.E染色方法对脊尾白虾的性腺发育及其组织结构进行观察。结果表明,脊尾白虾的雌性生殖系统由卵巢、输卵管及排卵孔组成。卵子发生经历了卵原细胞、卵黄合成前期卵母细胞、内源性卵黄合成期卵母细胞、外源性卵黄合成期卵母细胞,最后发育为成熟的卵母细胞。卵巢发育可分为增殖期、小生长期、大生长期、成熟期及产后恢复期。脊尾白虾雄性生殖系统由精巢、输精管及排精孔组成。精巢由生精小管构成,不同生精小管内精子发育可不同步。精子发生经历了精原细胞、初级精母细胞、次级精母细胞、精细胞,最后发育为精子。输精管可分为前、中、后输精管及末端壶腹,精荚在输精管中形成。     

长鳍篮子鱼繁殖季节性腺的组织学研究

采用常规组织切片法研究了长鳍篮子鱼繁殖季节各期卵巢和精巢的组织学结构特征。在所获得样本中,未发现Ⅰ期、Ⅱ期的卵巢和精巢。Ⅲ期卵巢以第Ⅲ时相卵母细胞为主,这时期液泡、卵黄颗粒出现,同时含有少量Ⅰ时相和Ⅱ时相的卵母细胞;Ⅳ期卵巢以第Ⅳ时相卵母细胞为主,Ⅳ时相晚期卵母细胞开始出现油球,细胞核偏移和变形,放射带明显;Ⅴ期卵巢的卵细胞游离,卵细胞的外层分别有胶膜、放射带和质膜;Ⅵ期卵巢主要由Ⅲ时相卵母细胞和大量的空滤泡外膜组成。长鳍篮子鱼精巢为辐射型,精巢内生殖细胞分为初级精母细胞、次级精母细胞、精子细胞和精子,各期生殖细胞和支持细胞组成了精小管,同一精小管中的生精细胞发育不同步。精子成熟后,充满整个精小管,完成发育过程。     

汤旺河花斑副沙鳅繁殖季节性腺发育组织学观察

运用组织切片技术研究汤旺河花斑副沙鳅Parabotia fasciata繁殖季节的性腺发育。结果表明:花斑副沙鳅性成熟系数在6月份最高,其中雄性群体的平均成熟系数为(1.26±0.28)%,雌性群体为(27.6±3.14)%;6~8月,花斑副沙鳅性成熟系数逐渐降低,推断其繁殖盛期在6月中旬~7月上旬。繁殖季节汤旺河花斑副沙鳅精巢发育处于精子形成阶段,精小囊相互融合,大量的精细胞向精子转变,排精后精巢内充满网状的结缔组织,染色后呈粉红色,精原细胞游离于囊腔内,精巢结构为典型的小叶型结构。花斑副沙鳅只有一侧卵巢,繁殖季节卵巢切面显示,主要由第Ⅰ、Ⅱ、Ⅲ(早期)、Ⅴ和Ⅵ5个时相卵母细胞构成,其中第Ⅰ、Ⅱ、Ⅲ(早期)和Ⅵ时相卵母细胞主要存在于产后恢复期的卵巢内,第Ⅴ时相卵母细胞存在于成熟卵巢内。成熟卵母细胞卵径为628.9~893.6μm,放射带厚度为13.69~20.486μm。依据卵巢切面组织学及性成熟系数月份变化,推测汤旺河花斑副沙鳅为一次性产卵鱼类。     

Reproductive cycle of yellow sea bream <Emphasis Type="Italic">Dentex tumifrons</Emphasis> in Wakasa Bay,the Sea of Japan off central Honshu

ABSTRACT:   To determine the spawning season and reproductive cycle of the yellow sea bream, Dentex tumifrons , in Wakasa Bay, in the Sea of Japan off central Honshu, seasonal change in the gonadosomatic index ( GSI ) was examined and histological observations of both ovaries and testes were conducted. The female GSI gradually increased from May, maintained a high level between August and October and then rapidly decreased. The histological observations of the ovary showed that vitellogenesis began from May and developed until September. Spent fish began to be observed in October and all the fish collected in December were spent. There was only one peak in the seasonal change in GSI of males and females in Wakasa Bay. Thus, yellow sea bream spawn only in autumn in Wakasa Bay. Differences in the reproductive cycle and number of spawning seasons were found between the two different latitudinal areas, Wakasa Bay and the East China Sea (low latitudinal area) where the spawning seasons were spring and autumn. An increase in water temperature is an important factor to initiate gonadal maturation (recrudescence) and is terminated by the short day-length in Wakasa Bay.     

中华锯齿米虾内外性征分化时间与特征

通过对中华锯齿米虾(Neocaridina denticulate sinensis)早期性腺发育的组织学观察和第一腹肢的形态学观察,研究了其雌雄性腺分化的时间、分化过程中生殖细胞及第一腹肢变化的特征。实验结果表明,卵原细胞出现在孵化后发育12～18 d的幼体,此时期所有幼体的第一腹肢内肢均具有刚毛。孵化后发育18～22 d幼体的性腺出现了两种类型:类型1和类型2,类型1具有典型的卵巢特征,之后沿着卵巢发育方向分化;类型2兼备精巢和卵巢的一些特征,之后沿着精巢发育方向分化。精巢的分化要经过一个类卵巢的阶段,在该时期还出现了雄性生殖肢,其第一腹肢内肢刚毛数量减少且为卵圆形,而雌性第一腹肢的内肢在形状与刚毛数量上无明显变化,雄性生殖肢的出现与精巢的分化时间是一致的。     

白斑狗鱼新基因lincRNA-Abcc10的克隆、表达及敲除模型构建

长非编码 RNA (lncRNAs)作为一种新的表观遗传因子, 在研究鱼类性别决定和性别分化过程中具有重要的作用。本研究利用 RACE 技术, 对白斑狗鱼(Esox Lucius)性别特异性 lncRNAs 基因 Novel01784 进行克隆, 获得了全长共 1328 bp 的序列。在线预测工具 CPC2 和 CPAT 的分析结果显示, 该基因不具有编码蛋白质的能力。通过 Blast 比对分析将其定位在 9 号染色体, 且位于 sp2 和 Abcc10 基因之间, 根据其位置关系将其命名为 lincRNAAbcc10。从共定位分析结果显示, lincRNA-Abcc10 可能参与调控离子运输、卵泡细胞发育和线粒体活性, 其表达模式与部分邻近基因相似。其次, 对白斑狗鱼 lincRNA-Abcc10 3 个时期在 8 个组织中的 qRT-PCR 结果显示, 该基因在卵巢中特异性表达, 随着时间增长表达量逐渐下降, 说明其可能在卵巢早期分化过程中起着重要的作用。此外, 通过对白斑狗鱼 126 d 卵巢核质 RNA 分离, qRT-PCR 结果显示, lincRNA-Abcc10 分布于细胞质内, 可能以顺式或反式方式调控邻近基因的翻译水平。最后, 利用 CRISPR/Cas9 技术建立了 lincRNA-Abcc10 突变模型, 为进一步研究其基因功能奠定了基础, 本研究为 lncRNA 在鱼类性别分化的作用提供了新的思路。     

Estimation of gonadal status of Japanese female eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> by a single injection of salmon pituitary extract

ABSTRACT:   Cultivated eels often either lack a part of the ovary (abnormal) or do not have any gonadal tissue (completely sterile). These abnormalities may cause a failure of induction of sexual maturation of female eels. In order to effectively induce sexual maturation and obtain good quality eggs, female eels having normal ovaries should be selected before induction of maturation. The present study examined effects of injection of salmon pituitary extract (SPE) on serum estradiol-17β levels to estimate gonadal status of sexually immature Japanese eels. At the initial stage (just prior to SPE injection), serum estradiol-17β levels were low and no significant difference was observed among eels possessing the different gonad types (normal, abnormal, and sterile), suggesting that gonadal status cannot be estimated by the measurement of serum estradiol-17β levels. A single injection of SPE significantly increased serum estradiol-17β levels in eels possessing normal and abnormal ovaries, but not in sterile eels. Serum estradiol-17β levels significantly increased from 24 h and were high on day 3. These data indicate that eels possessing ovaries can be distinguished from sterile eels by measurement of serum estradiol-17β between 1 and 3 days after a single injection of SPE.     

Effects of estradiol-17β and 17α-methyltestosterone on gonadal sex differentiation in the F2 hybrid sturgeon, the bester

The effects of giving oral estradiol-17β (E₂) and 17α-methyltestosterone (MT) on gonadal sex differentiation in the F₂ hybrid sturgeon, the bester ( Huso huso female ×  Acipenser ruthenus male), are investigated. Giving E₂ at 10 μg/g diet to fish from 14 months until 31 months of age induced incomplete feminization and resulted in approximately 40% abnormal ovary development in which oocytes were observed without ovarian lamellar structures and gonadal shape was similar to normal testis. Giving MT at 25 μg/g diet for the same duration failed to induce masculinization, and resulted in approximately 30% undeveloped gonads even at 30–37 months of age. In contrast, E₂ and MT at only 1 μg/g diet given from 3 to 18 months of age was sufficient to induce feminization and masculinization, respectively. In these fish, feminization and masculinization were observed at 9 months, when most putative ovaries and testes were histologically distinguishable by the shape of the gonadal surface. These results indicate that sex reversal can be induced in these fish by hormone treatment that is started at 3 months age, before morphological differentiation occurs on the stroma of the gonads.     

The dynamics of gonad growth and ascorbate status in yellow perch, Perca flavescens (Mitchill)

Yellow perch, Perca flavescens (Mitchill), caught in the wild (Grand St Marys Lake, Ohio) and cultured in the laboratory were used to examine the relationship between gonad development and the concentration of ascorbic acid in ovaries, testes, liver and kidney. In wild fish the ovary had concentrations of ascorbate twice as high (75-102 μg g^-1) as the testis (28-48 μg g^-1), and when the differences in gonadosomatic indices between sexes were accounted for, females deposited 10-fold more of this essential nutrient in gonads than did males. Yellow perch cultured intensively throughout the gonadal recrudescence had shown a decrease of ascorbate concentration in the kidney, liver and ovary during winter, most likely due to the low dietary level of the vitamin. Diet enrichment with ascorbic acid resulted in a dramatic increase of ascorbate in ovary and other tissues. Incorporation of ascorbic acid may take place during the late vitellogenesis, and this vitamin deserves to be studied as a quality indicator in fish eggs.     

Histological observations of seasonal reproductive and lunar-related spawning cycles in the female honeycomb grouper Epinephelus merra in Okinawan waters

Seasonal reproductive and lunar-related spawning cycles of the female honeycomb grouper Epinephelus merra inhabiting Okinawan waters were examined by histological observation of ovaries. Gonadosomatic index (GSI) increased beginning in May and peaked in June. Histological observations revealed that many oocytes laden with yolk were present in the ovaries from May to August. From September to October, ovaries were occupied by immature oocytes. These results suggest that the reproductive season of E.   merra lasts for 4 months from May through August in Okinawan waters. When the fish were collected according to the lunar cycle, GSI increased with the approach of the full moon. Oocytes at various development stages were observed from the first quarter to the full moon. Fresh ovulatory follicles were present in the ovaries around the last quarter moon. These results suggest that E.   merra has a lunar spawning cycle and spawns between the full moon and the last quarter moon. Ovarian features of the fish collected around the last quarter moon were different among individuals; some fish had many oocytes at the tertiary yolk stage in the ovaries, while the ovaries of the others were occupied by the oocytes at the peri-nucleolus and the oil droplet stages. This observation suggests that a minor release of eggs occurs in this species before or after a major spawning lunar day. Oocytes at the migratory nucleus and the maturation stages were not observed in any ovarian samples. This may mean that maturation of oocytes is related to the spawning behavior of this species and makes rapid progress in the process of aggregation at the spawning sites.     

半滑舌鳎卵巢发育的组织学和形态数量特征研究

采用组织学方法系统研究了人工养殖条件下半滑舌鳎亲鱼卵巢的组织发育周年变化特征.结果表明,卵母细胞发育可分为6个时相,卵巢发育分为6期.卵巢不同发育时期都由不同类型的卵母细胞组成,半滑舌鳎为非同步分批多次产卵类型.周年发育过程中,6月龄前的鱼卵巢发育处于第Ⅰ期;9～12月龄卵巢可发育至第Ⅱ期;12月龄半滑舌鳎性腺可发育达到Ⅲ期,并一直保持至24月龄.3龄雌鱼达到性成熟,卵巢可发育至第Ⅳ期;随着水温和光周期的调控,卵巢发育进入成熟期(Ⅴ期),发生水合现象是卵母细胞成熟并准备排卵的信号;产卵结束后卵巢退化至第Ⅵ期,排出的卵母细胞退化吸收,以Ⅲ期卵巢越冬后,卵巢退化至Ⅱ期并持续至下一个繁殖周期.人工养殖条件下,卵巢内存在卵母细胞闭锁现象.性腺周年发育过程中,性腺指数(GSI)在性腺达到成熟期时达到峰值,产卵结束后性腺快速退化,GSI值各月份差异显著(P<0.05),亲鱼性腺重量在卵巢发育成熟过程中一直处于生长状态.亲鱼肝脏指数(HSI)在产卵前7～8月份(性腺达到成熟期前30 d左右)达到最大,表明7～8月份是性腺进行卵子发育、卵黄能量储备的重要时期,亲鱼在产卵前通过积极摄食储备体内能量,肝脏在卵母细胞卵黄积累方面具有重要的生理作用.肥满度(CF)值在繁殖盛期10～11月份达到最大,表现出与GSI类似的变化趋势.     

Reproductive biology of common silver biddy Gerres oyena in Okinawa Island of southern Japan

ABSTRACT:   Common silver biddy Gerres oyena were collected monthly between November 2002 and November 2005 in Okinawa Island of southern Japan. The seasonal reproductive cycle of this species was investigated using the gonadosomatic index (GSI) and through histological observation of gonads. Temporal variation in the GSI reflected gonadal development from histological observations; ovary development occurred from March to September and testes development occurred between March and August. Maximal development was observed in April and May in both sexes. While mean values for hepatosomatic and lipidosomatic indices decreased markedly in females relative to males during gonadal development, mean condition factors decreased with gonadal recrudescence in both sexes. The minimum standard length (SL) at sexual maturity was 89.7 mm for females and 81.4 mm for males and the size at which 50% of individuals were sexually mature was 104 and 92 mm SL in females and males, respectively. The power relationships of batch fecundity (BF) versus standard length (SL, mm) and body weight (BW, g) of females can be expressed as BF  = 46.7 × e^{1.64 SL} and BF  = 19 697 × e^{0.49 BW} , respectively.     

水温对半滑舌鳎性腺组织发育的影响

探讨了不同温度(19,21,23,25和27 ℃)对半滑舌鳎的性腺组织发育的影响。在受精后39～106 d对半滑舌鳎进行温度控制处理,通过石蜡组织切片,对半滑舌鳎早期性腺分化发育进行组织学观察。发现在19 ℃饲养条件下,受精后46 d(pfd46)半滑舌鳎性腺原始生殖细胞数量增加,有分化成卵巢的倾向;在该条件下,pfd82半滑舌鳎性腺开始分化为精巢。pfd82时,27 ℃条件下的幼鱼卵巢中卵原细胞数目大量增殖;精巢中精小叶多而密,结缔组织间隔清晰。pfd129时,27 ℃处理的幼鱼卵巢发育到Ⅱ期,其他温度处理组仍处于Ⅰ期卵巢,但随着温度的提高,卵巢中卵母细胞数目增多,且由散乱分布变为有序的排列在卵巢小叶中;精巢中贮精囊数量增多,囊腔中精子增多。pfd142时,高温处理组幼鱼性腺发育最快,其Ⅱ期卵巢中卵母细胞数目最多,精巢中后期生精细胞偏多,精子几乎分布到各个精小叶中,且贮精囊结构发达,内纳高密度精子,在周围输精管中也分布着精子。结果表明,温度影响了半滑舌鳎性腺发育程度,二者呈现线性关系,在适当温度范围内,性腺发育速度随着温度升高而加快。