粗唇(鱼危)精巢显微结构和精子发生的研究

采用光镜和透射电镜观察了粗唇(鱼危)(Leiocassis crassilabris)精巢结构和精子发生.光镜结果显示:粗唇(鱼危)的精巢各小叶由数个小囊组成.从第Ⅱ期到第V期时,初级精母细胞逐渐发育成为次级精母细胞、精子细胞和精子,第Ⅵ期为排精后退化时期.透射电镜观察粗唇(鱼危)的精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子的形态以及细胞核和线粒体等的结果表明:在精子发生各期中线粒体和细胞核有明显的变化;精原细胞时期细胞器较丰富,到精子细胞后期,细胞器的形态和数量发生了较大变化;随着精母细胞的发育,核质凝聚程度逐渐增强;粗唇(鱼危) 的精子具有椭圆的头部和复杂的中片,并具有由外膜折叠形成的波浪形的结构.     

外源性促性腺激素诱导日本鳗鲡精子发生和成熟的作用机制

用兔抗血清对抗促黄体素生成素受体（LHR）或称绒毛膜促性腺激素受体（CGR）和雄激素受体（AR）进行LHR和AR免疫组织化学定位,以揭示外源性促性腺激素（鲤脑垂体激素和hCG）诱发日本鳗鲡精子发生及其内分泌机制。结果表明，经过注射激素处理后的实验组与注射前的对照组相比较，其精巢发育和精子发生出现十分显著的变化。组织学切片观察显示,激素处理前鳗鲡精巢处于精原细胞增殖期，而两种激素混合注射后第10天，实验组可见精小叶中精原细胞的有丝分裂和初级与次级精母细胞的数量显著的增加。注射后第35天，靠近生殖上皮除有少量精原细胞外，精小叶中有大量初级精母细胞和次级精母细胞和少数精子细胞以及管腔中存在少量精子。在注射后第83天，日本鳗鲡完成了精子发生和精巢发育成熟以及释精。免疫组织化学染色结果进一步揭示，激素处理前，LH受体免疫活性分布在生殖上皮，显示强的免疫阳性反应；激素处理后，LH受体定位在Sertoli细胞和间质细胞以及精原细胞和初级与次级精母细胞的胞膜上，均显示强的免疫阳性反应。激素处理前，雄激素受体定位在生殖上皮和早期生精细胞的胞膜上；激素处理后，AR则定位在这些生精细胞的核或胞质，而精子细胞和精子显示免疫阴性反应。这些结果首次证明了这两种激素诱导鳗鲡精子发生和成熟的作用机制是通过LH受体和雄激素受体的介导。     

长薄鳅的精巢发育和精子发生

应用组织学和超微结构方法研究了长薄鳅(Leptobotia elongate Bleeker)精巢发育和精子发生。结果显示:长薄鳅精巢一对,其发育过程可划分为6个时期,Ⅰ期性腺肉眼无法分辨雌雄。组织学结构显示长薄鳅的精巢属于小叶型,根据发育过程中细胞的大小、核仁多少、细胞核和细胞质浓缩的程度将精子发生划分为6个时相。超微结构下精原细胞分为初级精原细胞和次级精原细胞,成熟精子分头部、中片和尾部3个部分,尾部有侧鳍,具"9+3"式微管结构。     

粗唇鮠精巢显微结构和精子发生的研究

采用光镜和透射电镜观察了粗唇鮠(Leiocassis crassilabris)精巢结构和精子发生。光镜结果显示:粗唇鮠的精巢各小叶由数个小囊组成。从第Ⅱ期到第V期时,初级精母细胞逐渐发育成为次级精母细胞、精子细胞和精子,第Ⅵ期为排精后退化时期。透射电镜观察粗唇鮠的精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子的形态以及细胞核和线粒体等的结果表明:在精子发生各期中线粒体和细胞核有明显的变化;精原细胞时期细胞器较丰富,到精子细胞后期,细胞器的形态和数量发生了较大变化;随着精母细胞的发育,核质凝聚程度逐渐增强;粗唇鮠的精子具有椭圆的头部和复杂的中片,并具有由外膜折叠形成的波浪形的结构。     

广东鲂性腺发育组织学研究

利用组织学方法,对珠江下游野生广东鲂(Megalobrama terminalis)性腺进行了显微观察和研究,描述了其卵巢、精巢的结构特征以及各类型生殖细胞形态。结果显示,广东鲂属于一次性产卵类型,卵巢发育类型为部分同步型。其卵巢为细线或条状,卵巢发育经历卵原细胞期、单层滤泡细胞期、卵黄泡期、卵黄充满期、成熟期和退化期。卵细胞发生经历3个阶段:卵原细胞、初级卵母细胞和次级卵母细胞;卵巢初级卵母细胞中核仁外排现象出现在第2时相末期至第3时相早期;精巢呈线状或直棒状,精巢为叶状型结构。精细胞发生经历5个阶段:精原细胞、初级精母细胞、次级精母细胞、精子细胞及精子;繁殖季节精子充满于精小叶内,精小囊消失。     

双须骨舌鱼性腺发育的组织学观察

通过组织学研究方法,分别对双须骨舌鱼Ⅱ~Ⅴ期的卵巢及Ⅱ~Ⅵ期精巢的形态结构、特征及生殖细胞变化进行描述。结果显示,Ⅱ时相卵母细胞的细胞核较大,约为细胞体积的一半;Ⅲ时相卵母细胞出现卵黄斑及卵黄颗粒;Ⅳ时相卵母细胞中卵黄迅速积累且细胞膜出现褶皱;Ⅴ时相卵母细胞膜表面的褶皱消失。雄性双须骨舌鱼精巢发育的Ⅱ期,只含有初级精母细胞。Ⅲ期精巢分布有初级和次级精母细胞,精小管形成且小管中充满精细胞;Ⅳ期精巢主要含有初级和次级精母细胞以及精原细胞,精小管中出现空腔;Ⅴ期精巢中包含次级精母细胞、精原细胞和精子细胞,精小管中的空腔增大;Ⅵ期精巢则显示出高度血管化和结缔组织增多等特点。本研究将为双须骨舌鱼人工繁殖提供参考。     

单环刺螠精巢年周期发育及精子发生

采用组织学方法观察了单环刺螠(Urechis unicinctus)精巢的形态结构及其年周期发育,并利用超微技术观察了精子的发生过程。结果表明,单环刺螠的精巢呈条带状,位于虫体尾部,两端通过肌束分别与呼吸肠管壁外侧和体壁内侧相连。精巢由生精细胞团与结缔组织组成,结缔组织位于精巢中央,精原细胞团分布于结缔组织外周,精母细胞团散布于结缔组织内,各级精母细胞脱离精巢落入体腔液中,历经精母细胞、精细胞,最后形成精子脱离细胞团进入肾管。根据精巢的组织学特征,将单环刺螠的精巢年周期发育划分为增殖期、生长期、成熟期、排放期和休止期5个时期。光镜和电镜下观察了精巢和体腔液中精子发生各时期的结构变化,包括细胞核形态的变化、顶体发生以及尾部的形成。本研究揭示了单环刺螠的精巢发育特点、精子发生历经的场所和分化特征。     

西埔湾黄鳍鲷精子发生和形成

本文在超微结构水平上,着重研究了西埔湾黄鳍鲷精子发生和形成过程中各级精细胞形态结构的变化、线粒体和高尔基体等细胞器的演变特点以及精巢内一些体细胞的形态结构与功能。研究结果证实了黄鳍鲷雄性性腺能在半封闭的西埔湾内发育成熟。精子发生和形成可分为原始精原细胞、精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子形成等6个发育阶段。成熟精子由头部、中段和尾部组成,头部近圆形,细胞核呈马蹄形,核内染色质高度浓缩,核上方不具顶体。尾部轴丝为“9×2+2”结构。     

二倍体和三倍体皱纹盘鲍精子发生过程的超微结构

比较了二倍体和三倍体皱纹盘鲍精子发生过程中细胞和细胞器的超微结构变化。结果表明，二倍体皱纹盘鲍的精子发生经历了精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子5个阶段。其形态结构发生了一系列变化，主要包括：核染色质浓缩、线粒体的发达与融合、顶体形成和胞质的减少。三倍体皱纹盘鲍各种生精细胞的直径和核径均大于二倍体，精原细胞结构与二倍体相似；初级、次级精母细胞的胞质中，除溶酶体外，线粒体、内质网等细胞器少于二倍体，线粒体大小与二倍体没有差别，但形态不典型，层状嵴不发达；三倍体皱纹盘鲍的精子发生停滞在精子细胞阶段，表现出各种畸形状态，很多趋于解体，没有发现成熟精子。     

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。     

青虾幼虾发育时期性腺发育组织学研究

采用组织切片及类固醇激素含量测定等方法,研究青虾Macrobrachium nipponense幼虾发育第1~31d的精巢、卵巢及促雄腺发育的起始时间、发育过程及成熟时间。结果表明:青虾幼虾发育到第10d(PL10)时促雄腺呈索状结构开始发育,随后经历增殖期(PL10)、合成期(PL13)和分泌期(PL19)3个发育阶段发育成熟,形成完整的促雄腺。青虾精巢和卵巢均在幼虾发育第13d(PL13)开始发育,精巢形成不规则排列的精原细胞,而卵巢生殖上皮开始分化为椭圆或多边形的卵原细胞。精巢经精原细胞期(PL13)、精母细胞期(PL16)、精细胞期(PL19)和精子期(PL22)4个发育阶段成熟,此时成熟的精巢生精小管内充满成熟的精子。卵巢经卵原细胞期(PL13)、初级卵母细胞期(PL16)、次级卵母细胞期(PL16)和卵子期(PL19)4个时期发育成熟,此时卵巢充满成熟卵子。从PL1到PL22,睾酮分泌量逐渐增加,维持在稳定水平至PL25后逐渐下降;从PL1到PL19,17β-雌二醇的分泌量逐渐增加,维持在稳定水平至PL25后逐渐下降。本研究显示:促雄腺与精巢发育过程均持续10d,但促雄腺早于精巢3d开始发育和成熟,这可为性别和生殖相关基因的筛选及性别调控机制的研究提供参考。     

Testicular responsiveness to gonadotropic hormonein vitro and Leydig and Sertoli cell ultrastructure during pubertal development of male African catfish (Clarias gariepinus)

The gonadotropin (GTH)-stimulated testicular androgen secretionin vitro and the ultrastructure of Leydig and Sertoli cells was studied during the pubertal development in male African catfish. Testicular weight increased from less than 1 mg in the ninth week of age to nearly 600 mg in the 28th week. Immature testes (stage I: spermatogonia) were highly sensitive to GTH and secreted very high amounts of androgens per mg of tissue. The secretion per mg tissue decreased gradually in stages II (spermatogonia and spermatocytes) and III (spermatogonia, spermatocytes, and spermatids), but precipitously in stage IV (all germ cell stages, including spermatozoa). However, due to the testicular weight gain, the total androgen output per pair of testes increased slightly in stage III and strongly in stage IV. The sensitivity to GTH decreased with the appearance of haploid germ cells in stage III. Leydig cells but not Sertoli cells showed the ultrastructural characteristics of steroid producing cells. Leydig cell morphology did not change in stages I–III, while in stage IV, more smooth endoplasmic reticulum was present. The ultrastructural characteristics of Sertoli cells did not change prominently. Thus, spermatogonial multiplication and spermatocyte formation takes place when the testicular steroidogenic system is highly active and responsive to GTH; whereas the differentiation of haploid germ cells is accompanied by a reduced responsiveness to GTH and by the secretion of several-fold lower androgen amounts per mg of tissue.     

瘤背石磺两性腺周年发育规律的研究

研究了江、浙、沪地区瘤背石磺（Onchidium struma）两性腺周年发育规律。结果显示,瘤背石磺雌雄同体,雄性先熟,生物学最小型为体长2.81cm,体质量1.81g。其两性腺发育为1年1个周期,发育经历增殖期、精子发生期、精卵同步发生期、排放期和休止期。其中排放期又分精子排放、精卵排放和卵子排放,体现了精卵发生的异时性。精子的发生包括精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子;卵子的发生包括卵原细胞期、无卵黄期、卵黄形成期、卵黄形成后期和成熟期。成熟系数周年呈明显单峰分布,峰值出现在7～9月间;蛋白腺系数、卵黄腺系数与成熟系数具有一定的相关性,蛋白腺系数基本随着成熟系数变化而变化,卵黄腺系数随着成熟系数变化的过程中出现3个峰值,分别在6、8和12月。     

长鳍吻鮈性腺发育的初步观察

采用常规石蜡切片技术对野生长鳍吻鮈(Rhinogobio ventralis(Sauvage et Dabry))性腺进行了组织学观察,描述了其各类型生殖细胞形态及卵巢、精巢的分期特征。结果显示:长鳍吻鮈卵巢为细线状或扁条状,精巢为细线状或直棍状;卵细胞发生经历了卵原细胞、初级卵母细胞和次级卵母细胞3个阶段,精细胞发生经历了精原细胞、初级精母细胞、次级精母细胞、精子细胞及精子5个阶段;性腺发育分为6个时期,卵巢中第Ⅱ时相晚期至Ⅲ时相早期的初级卵母细胞中出现核仁外排现象;同一时期卵巢中存在多种时相的卵母细胞,故推测其卵巢发育类型为部分同步型,精巢为小叶型结构。     

The urogenital papillae as an indicator of sexual maturity in male African catfish,Clarias gariepinus, (Burchell, 1822)

The present study investigated a non‐invasive method based on macroscopic morphological features of male Clarias gariepinus for the assessment of sexual maturity stage. As African catfish cannot be stripped for semen as in most other fish species; they have to be killed to obtain semen from the testes. This method could be used to ensure that males used to obtain sperm are sexually mature to increase the concentration and quality of mature sperm for egg fertilization in the production of C. gariepinus. Morphological features including the length and width of the urogenital papillae (UGP), an external secondary sexual characteristic of C. gariepinus (n = 89) were measured and calculated as a percentage of total body length. The urogenital papillae length (L_UGP) and width (W_UGP) index of each fish were used and compared with the macroscopic and microscopic maturity stage of the testes. The results revealed that although positive, a significant correlation does not exist between the L_UGP or W_UGP and maturity stage of the testes. Therefore, although this procedure would be valuable as an initial non‐invasive assessment of maturity, it does not provide an accurate or reliable indication of the maturity stage of the fish.     

养殖中华鲟性腺发生与分化的组织学研究

用组织切片方法研究人工繁育中华鲟的性腺发生及两性分化过程。中华鲟出膜后3d，原始生殖细胞以单细胞的形式存在于肾管区腹下方。出膜后11d，生殖褶形成，到2月龄和7月龄时，性腺中分别出现血管和脂肪组织，直到8月龄性腺均处于两性未分化状态，划分为0期。9月龄(体重0．6～1．1kg)，性腺出现组织学水平上的两性分化，性腺划入I期。I期性腺中，精(卵)原细胞进行有丝分裂。以初级精(卵)母细胞出现作为Ⅱ期开始的标志，精巢和卵巢分别于1．8～2．2年龄(1.55～5．6kg)和2．5～3．0年龄(3．1～8．2kg)进入Ⅱ期。3年龄以后，性腺发育分化陆续达到肉眼可分辨性别的程度，5～5．6年龄(18～35．5kg)时，所有的性腺能肉眼区分雌雄，但精巢和卵巢仍都处于Ⅱ期，其中卵母细胞的卵径为60～240μm。     

The effect of whole body lipid on early sexual maturation of 1+ age male chinook salmon (Oncorhynchus tshawytscha)

Early sexual maturation of male chinook salmon (maturation 1 to 4 years prior to females in the same age class) results in reduced effectiveness of stock enhancement programs and a financial loss to the salmon farming industry. Previous studies in Atlantic salmon have shown that the age of maturity in males is affected by growth and/or body energy stores, but the relative roles of these two factors are not well understood. Therefore, an experiment was designed to determine when spermatogenesis was initiated, to characterize the endocrine changes during the onset of puberty in male salmon, and to determine if the level of whole-body lipid affects the incidence of early male maturation in a wild stock (Yakima River) of 1+ spring chinook salmon. Fry were fed a commercial diet from February until August and were then divided into groups of 320 fish (mean weight, 5.6 g) and fed one of five experimental diets (two replicate groups/diet) containing 4%, 9%, 14%, 18% or 22% lipid and 82%, 77%, 73%, 69%, or 65% protein for 13 months. Fish were reared on natural photoperiod and ambient temperature (6°C to 16°C), and pair-fed to a level based on the tank with the lowest feed consumption. Fish were weighed monthly and sampled to determine body composition, pituitary follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, plasma insulin-like growth factor I (IGF-I) levels, and stage of gonadal development.

Throughout the experimental period the mean fish weight was similar among treatment groups. However, from December through the end of the experiment in the following September, maturing males were significantly larger than nonmaturing fish. Initial lipid levels in 0-age experimental fish were near 6%, which is similar to wild fish of the same stock and age captured in the Yakima River during August. Fish fed diets containing more than 4% lipid increased in whole-body lipid content during the first 2 months of feeding and then maintained at relatively constant levels during the course of the experiment. Whole-body lipid levels for the dietary treatment groups averaged 5.6%, 7.1%, 8.2%, 9.4%, and 9.6% from October through the following September.

Based on histological examination of the testes of experimental fish, type B spermatogonia and primary spermatocytes were first observed in some of the yearling males during November. These were designated maturing males. Pituitary FSH levels were significantly higher in maturing than nonmaturing males at this time and for the remainder of the study. Pituitary FSH levels increased as spermatogenesis proceeded in maturing fish, whereas pituitary LH levels increased in maturing 1+ males only during July and August, when testes were in late stages of spermatogenesis and in September during spermiation. Plasma IGF-I levels were significantly higher in maturing males than nonmaturing fish from December through the end of experiment. Since maturing males were significantly larger than nonmaturing fish of both sexes from December through September, the difference in IGF-I levels could be due to differences in growth or due to maturation.

The percentage of maturing males was significantly influenced by whole-body lipid, increasing from 34% in fish fed the 4% lipid diet to 45% in fish fed the 22% lipid diet. These data suggest that whole-body lipid levels influenced the incidence of maturation of male spring chinook salmon. In addition, both endocrine and histological indicators suggest that maturation was initiated in males approximately a full year prior to the time the fish will spawn.