秦岭北坡中国林蛙精巢显微结构的年周期变化

用光镜观察了秦岭北坡中国林蛙(Ranachensinensis)精巢显微结构的年周期变化,结合精巢系数的变化探讨其生殖规律。结果显示,秦岭北坡中国林蛙的生精周期属于非连续型。精巢系数的变化与精子发生的活动周期相一致。精子发生从每年5月开始,翌年4月结束,历时1年。生精周期可划分为5个时期。Ⅰ期,精原细胞增殖期,5～7月,精巢系数最小,精原细胞进行有丝分裂;Ⅱ期,精母细胞成熟分裂期,8～9月,精巢系数最大,精原细胞、精母细胞和精子细胞在生精小管内共存;Ⅲ期,精子形成期,9～1 0月,精子细胞变态形成精子;Ⅳ期,成熟精子贮存越冬期,1 1月至翌年2月,成熟精子贮存在生精小管中;V期,精子排放期,翌年3～5月,精巢系数显著下降,成熟精子从生精小管脱离,通过输精管道排出体外。     

中国林蛙消化管胚后发育的组织学观察

用解剖和组织切片技术对中国林蛙Rana chensinensis蝌蚪(23～46期)消化管的形态和显微结构变化进行观察比较.结果 表明,在中国林蛙幼体的胚后发育过程中,消化管长度从28期的37.0 mm逐渐增加至39期的95.3 mm,在变态过程中又缩短至22.0 mm.胃从42期开始膨大, 46期在位置和形态上与成蛙相似.消化管粘膜上皮逐渐完善,粘膜下层和肌层逐渐出现并加厚.胃腺、小肠腺以及杯形细胞在42～43期出现.说明林蛙消化管的组织结构特征与变态发育过程中食性的变化相适应.     

原癌基因FOS蛋白、雌二醇及其受体在中国林蛙不同时期精巢中的表达

用免疫细胞化学方法检测了原癌基因FOS蛋白、17β-雌二醇（E₂）和雌激素受体（ER）在中国林蛙生精周期中不同时期精巢内的表达定位。结果显示：在中国林蛙生精周期的Ⅰ—Ⅴ期，E₂和ER在精原细胞、精母细胞、精子细胞、精子、支持细胞和间质细胞内均有表达。在不同时期的精巢中，E₂和ER在生精细胞的定位具有一致性：在Ⅱ—Ⅲ期，精子细胞的E₂和ER阳性表达最强；在Ⅲ期，精子中E₂和ER阳性反应强度显著高于Ⅳ—Ⅴ期（P＜0.01)。在生精周期的Ⅰ—Ⅴ期，支持细胞中，E₂和ER表达强度经历由强减弱再增强的变化过程。在生精周期的Ⅲ期，间质细胞中E₂和ER阳性反应强度高于其他各期。除精子外的生精细胞，支持细胞和间质细胞内均有FOS阳性反应，其表达强度呈现阶段性变化。     

原癌基因FOS蛋白、雌二醇及其受体在中国林蛙不同时期精巢中的表达

用免疫细胞化学方法检测了原癌基因FOS蛋白、17β-雌二醇(E2)和雌激素受体(ER)在中国林蛙生精周期中不同时期精巢内的表达定位。结果显示:在中国林蛙生精周期的Ⅰ—Ⅴ期,E2和ER在精原细胞、精母细胞、精子细胞、精子、支持细胞和间质细胞内均有表达。在不同时期的精巢中,E2和ER在生精细胞的定位具有一致性:在Ⅱ—Ⅲ期,精子细胞的E2和ER阳性表达最强;在Ⅲ期,精子中E2和ER阳性反应强度显著高于Ⅳ—Ⅴ期(P<0.01)。在生精周期的Ⅰ—Ⅴ期,支持细胞中,E2和ER表达强度经历由强减弱再增强的变化过程。在生精周期的Ⅲ期,间质细胞中E2和ER阳性反应强度高于其他各期。除精子外的生精细胞,支持细胞和间质细胞内均有FOS阳性反应,其表达强度呈现阶段性变化。     

革胡子鲇卵巢在第一次性周期内分化与发育的研究

用光镜和电镜研究了革胡子鲇（Ｃｌａｒｉａｓｌａｚｅｒａ）原生殖细胞的起源迁移,卵巢在第１次性周期内的分化与发育以及各发育时期卵母细胞的超微结构。原生殖细胞起源于内胚层,有固定的迁移路线,进入生殖嵴后,生殖嵴进一步分化,出现卵巢腔分化成卵巢,卵母细胞在第１次性周期内的发育可分成６个时相,描述了各时相卵母细胞的显微结构与超微结构。同时,叙述了卵母细胞中卵黄发生的形态形成。     

乌梢蛇卵泡不同发育期颗粒细胞的显微结构变化

采用光学显微镜主要观察了乌梢蛇(Zaocys dhumnades)卵泡发育过程中颗粒细胞的显微结构变化.结果表明,乌梢蛇的滤泡前体细胞在形态、大小与嗜色性上均与生殖基的表面上皮相似;在原始卵泡期,卵母细胞周围的滤泡前体细胞围绕成一圈;在颗粒层细胞期的时期I至时期Ⅲ,颗粒细胞分化为由小细胞、梨形细胞与中间型细胞构成的异型颗粒细胞;在时期Ⅳ,这些异型颗粒细胞又转变成为只有小细胞的同型颗粒细胞.乌梢蛇卵泡颗粒细胞来源于生殖基的表面上皮,其发育特征是首先由同型发育成为异型,再由异型转变为同型颗粒细胞,具有同源异型的特征.     

昆明地区昭觉林蛙生殖及附属器官的年周期变化

解剖2006年4—12月自昆明东北郊采集到的体长大于40.0mm的昭觉林蛙(Rana chaochiaoensis)标本73只(♀39,♂34),测量其体长、体重、肝重、脂肪体重、卵巢重、精巢重、输卵管重和直径、卵径等。以体长(或体重)为协变量做协方差分析(ANCOVA),结果显示,输卵管直径、卵径,雌性的肝重、脂肪体重、卵巢重和输卵管重和雄性的精巢重在年周期内差异极显著。相关性检验显示,雌性卵巢重分别与肝重、输卵管重、输卵管直径、卵径均显著正相关,即它们的发育完全同步,但与脂肪体重不相关。各月雌体卵巢重和雄体精巢重的发育极显著正相关,即雄性与雌性的性腺发育同步。据解剖、检验分析及野外观察结果分析证明,昆明地区昭觉林蛙的繁殖时间在10月完成,且一年只有一个繁殖期。脂肪体在雌性个体生殖腺发育过程中起供能作用。在卵巢发育过程中,肝供能作用不明显,但在雌性个体抱对和产卵过程中起供能作用。在雄性性腺发育的过程中,肝和脂肪体的供能作用不如雌性明显,耗能极低。可见昭觉林蛙雄性个体的生殖投入少于雌性个体。一些雌性个体推迟冬眠时间,主要是为觅食和补充能量,以便安全越冬。此外,新生昭觉林蛙蝌蚪期在冬季度过,随后的发育和变态面临旱季缺水环境。所以,加强对昭觉林蛙产卵高峰期、蝌蚪越冬期和渡过干旱缺水期等生活周期中脆弱环节的环境保护,有利于种群的延续。     

乌梢蛇精巢显微结构的年周期变化

用光镜观察了乌梢蛇(Zaocys dhumnades)精巢显微结构的年周期变化,并结合精巢重量、精巢体积及精巢系数的年周期变化探讨了其生殖规律。结果表明,乌梢蛇精巢重量、精巢体积、精巢系数、曲细精管直径、生精上皮厚度及组成均具有明显的季节性变化。据此将乌梢蛇精巢的年周期活动划分为6个时期,其精子发生属于非连续型,生殖周期的类型属于交配后型。用精巢系数的年周期变化作为参数来判定精子发生进程是可靠的。     

昆明地区昭觉林蛙生殖及附属器官的年周期变化

解剖2006年4—12月自昆明东北郊采集到的体长大于40.0 mm的昭觉林蛙(Rana chaochiaoensis)标本73只(♀39, ♂34),测量其体长、体重、肝重、脂肪体重、卵巢重、精巢重、输卵管重和直径、卵径等。以体长(或体重)为协变量做协方差分析(ANCOVA),结果显示,输卵管直径、卵径,雌性的肝重、脂肪体重、卵巢重和输卵管重和雄性的精巢重在年周期内差异极显著。相关性检验显示,雌性卵巢重分别与肝重、输卵管重、输卵管直径、卵径均显著正相关,即它们的发育完全同步,但与脂肪体重不相关。各月雌体卵巢重和雄体精巢重的发育极显著正相关,即雄性与雌性的性腺发育同步。据解剖、检验分析及野外观察结果分析证明,昆明地区昭觉林蛙的繁殖时间在10月完成,且一年只有一个繁殖期。脂肪体在雌性个体生殖腺发育过程中起供能作用。在卵巢发育过程中,肝供能作用不明显,但在雌性个体抱对和产卵过程中起供能作用。在雄性性腺发育的过程中,肝和脂肪体的供能作用不如雌性明显,耗能极低。可见昭觉林蛙雄性个体的生殖投入少于雌性个体。一些雌性个体推迟冬眠时间,主要是为觅食和补充能量,以便安全越冬。此外,新生昭觉林蛙蝌蚪期在冬季度过,随后的发育和变态面临旱季缺水环境。所以,加强对昭觉林蛙产卵高峰期、蝌蚪越冬期和渡过干旱缺水期等生活周期中脆弱环节的环境保护,有利于种群的延续。     

卵胎生硬骨鱼褐菖You精巢的周期发育

研究了卵胎生硬骨鱼褐菖Ｙｏｕ（Ｓｅｂａｓｔｉｓｃｕｓ ｍａｒｍｏｒｄｔｕｓ）的精巢结构和生殖周期。褐菖Ｙｏｕ精巢属于小叶型。每年８～９月,精巢处于精原细胞增殖期。初级精原细胞分裂增殖,产生次级精原细胞。后者和支持细胞组成精小囊。１０月～翌年、月进入精子发生期。精小囊中的生殖细胞进一步发育,逐渐形成精子。２～７月是精子退化吸收期,精巢中仅有初级精原细胞和残余的精子。在生殖季节,精子经由输出管和输精管     

Immunoreactivities of AR,ERα, ERβ and aromatase in the nuptial pad of Chinese brown frog (Rana dybowskii) during pre-hibernation and the breeding period

There is a prominent local raised pad called nuptial pad on the forelimb of Chinese brown frog (Rana dybowskii), which is hypothetically concluded as an enhancement of the grip and a spreader of pheromone during the amplexus. In this study, we investigated the immunolocalization and protein expression levels of androgen receptors (AR), estrogen receptor α (ERα), ERβ and aromatase in the nuptial pad of R. dybowskii during pre-hibernation and the breeding period. Histologically, the annual development of the nuptial pad in R. dybowskii is manifested as the larger area of specialized mucous gland and the longer length of papillary epidermal projection during the breeding period. AR, ERα, ERβ and aromatase are present in the stratum granulosum, stratum spinosum, stratum basale and the secretory portion of specialized mucous glands during both periods. Western blotting results confirmed that AR, ERα and ERβ protein levels are higher during pre-hibernation than those during the breeding season. These results suggest that nuptial pad is the direct target organ of androgen and estrogen. Androgen may participate in the regulation of annual development and glandular function of nuptial pad, and estrogen may play an endocrine, autocrine or paracrine role during pre-hibernation and the breeding period.Key words: Androgen receptor, aromatase, estrogen receptor, nuptial pad, Rana dybowskii.     

钉螺体内日本血吸虫尾蚴发育期的形态及其扫描电镜观察

本文通过活体标本、连续切片、全封标本及扫描电镜观察了大陆品系日本血吸虫Schistosomajaponicum子胞蚴体内尾蚴发育期的形态。日本血吸虫尾蚴发育分为五期,即胚细胞期、胚球期、尾蚴雏体期、成熟前期和成熟期。与曼氏血吸虫尾蚴发育进行比较,日本血吸虫尾蚴在胚球期无极化现象;尾蚴雏体期的尾芽形态多非圆球形,尾叉形成较早。钻腺及焰细胞均在此期发生,较曼氏血吸虫尾蚴早。除了上述头器、头腺、钻腺、焰细胞及消化器官进一步发育外,体内散在胚细胞结集为生殖始基,体表感觉乳突分化与体尾肌细胞的发育,尾蚴从成熟前期过渡到成熟期。 电镜首次原位观察尾蚴发育在子胞蚴育腔内的自然状态。尽管偶尔可见到胞蚴体壁与胚元之间某些结构联系,但大部份的胚元各自独立混杂在育腔之中。此外尚注意到尾蚴体棘发生在成熟前期,但其密度较成熟尾蚴小,而其大小却比成熟尾蚴的大。尾蚴头器上感觉乳突和围褶可能先于内部腺体细胞的分化。     

Annual changes in germinal epithelium determine male reproductive classes of the cobia

Five reproductive classes of cobia Rachycentron canadum , caught along the Gulf of Mexico and the south-east Atlantic coast of the U.S.A., are described during the annual reproductive cycle. These are based upon changes in the testicular germinal epithelium and the stages of germ cells that are present: early maturation, mid maturation, late maturation, regression and regressed. During early maturation, the germinal epithelium is continuous from the testicular ducts to the periphery of the testis and active spermatogenesis occurs throughout the testis. In mid maturation, the germinal epithelium near the ducts becomes discontinuous, but it remains continuous distally. In late maturation, a discontinuous germinal epithelium extends all along the lobules to the testicular periphery; lobules are swollen with sperm and there is minimal spermatogenesis. The regression class is characterized by a discontinuous epithelium throughout the testis, sperm storage and widely scattered spermatocysts. Spermatogonial proliferation also occurs along the lobule walls and at the periphery of the testis. In regressed testes, spermatogonia exist only in a continuous or discontinuous germinal epithelium, although residual sperm are nearly always present in the lobules and ducts. The presence or absence of sperm is not an accurate indicator of reproductive classes. At the periphery of the testis in the regression and regressed classes, the distal portions of lobules elongate as cords of cells containing spermatogonia and Sertoli cells. All reproductive classes can be identified in paraffin sections, although plastic sections provide better resolution. Using maturation classes defined by changes in the germinal epithelium to describe testicular development and spermatogenesis gives a more accurate picture than does using the traditional terminology.     

Froghood: Postmetamorphic development of the rock river frog Thoropa miliaris (Spix, 1824) (Anura,Cycloramphidae)

Thoropa miliaris is a frog species endemic to the Brazilian Atlantic Rainforest, inhabiting wet rocky habitats. Males of this species present nuptial excrescences, which are keratinized spines on the surface of some fingers and internal metacarpal tubercles. Although these spines are usually associated with male adulthood, no details of their development are available. To investigate the sequence of spines' appearance, we correlated the number of spines with size and age, which was estimated by skeletochronology. We studied individuals collected from Rio de Janeiro, Brazil. There was significant correlation between size of individuals and number of spines and between age and number of spines in the callosities. The first spines appeared in metacarpal–phalangeal articulation of finger II, when specimens were 1 year old. The estimated growth curve did not show stabilization at any age. We considered the degree of development of the nuptial excrescences as a proxy for sexual maturity and so individuals may already be sexually active by 1 year of age. Estimated longevity of T. miliaris is similar to that estimated for other tropical species of frogs. Although growth in anuran species is considered to be undetermined, in T. miliaris, it seems to fit either undetermined or determined models.     

Immunolocalization of PTHrP in prepubertal and pubertal testis of European bison

The present study deals with immunohistochemical localization of PTHrP in prepubertal and pubertal testis of European bison. PTHrP immunoreactivity was observed in germinal cells in the testis of both prepubertal and pubertal animals. In calves, PTHrP was found in germinal cells, in seminiferous tubules lacking the lumen. The reaction was strong and regularly distributed within the cytoplasm. In adult animals, the reaction showed differentiation in spermatogenic cells. Some cells were strongly and diffusely stained, others exhibited weaker reaction of granular pattern. Sertoli cells and Leydig cells were PTHrP-negative in calves and adult animals.     

Cultivation in vitro of the testis cord of the newt, Triturus pyrrhogaster

Testis cords of Triturus pyrrhogaster were cultivated in vitro on (a) medium with chick embryo extract and calf serum, (b) medium with newt gonad extract, (c) Trowell 's medium T8 and (d) liquid synthetic medium 199. Of the four media utilized, medium 199 gave the best result for long-term maintenance of the normal histological structures of the testis cords. Addition of insulin (5 μ/ml) to medium 199 resulted in a remarkable improvement for the maintenance of the testis cord and the migration of columnar cells of the peritoneal epithelium into the primordial germinal tissue occurred as in the intact testis of this animal. Trowell 's medium T8 was proved inadequate. Medium with chick embryo extract and calf serum retained most of the germ cells healthy but caused gradual decrease in height of the columnar cells. Testis cords cultivated on the same medium in combination with Xenopus testis maintained normal histological structure for 18 days, whereas, those kept in contact with Xenopus ovary showed involution within the same period. Newt testis extract brought about transformation of somatic elements of the germinal tissue into fibroblastlike cells which was followed by the disintegration of germ cells. Ovary extract did not cause selective destruction on the somatic or germinal elements.     

Seasonal lectin binding variations of thumb pad in the frog (Pelophylax ridibundus)

The thumb pad is one of the most common secondary sexual characteristics in frogs. Although it is known that amphibian skin has affinity for several lectins, there is no report regarding lectin‐binding affinity of the thumb pad or its structural components. This study investigated localization and seasonal variation of specific carbohydrate moieties of glycoconjugates in both the epidermal and dermal components of the frog thumb pad at the light microscopic level using lectin histochemistry. The study consisted of four seasonal groups of the frog species, Pelophylax ridibundus (Synonym of Rana ridibunda): active, prehibernating, hibernating and posthibernating. Four horseradish peroxidase conjugated lectins were employed. It was found that dolichos biflorus agglutinin (DBA), wheat germ agglutinin (WGA), and ulex europaeus (UEAI) gave positive reactions in both epidermal layers and breeding glands. These three lectins bound specific secretory cells in the breeding glands, and the distribution of the cells and epithelial lectin reactions exhibited seasonal changes. In addition, UEA‐I and peanut agglutinin (PNA) showed an affinity in granular glands and the granular zone of mixed glands. Generally, epidermal lectin binding showed dense affinity during the posthibernation period. DBA, UEA‐I, and WGA‐specific cells in the mucous gland decreased gradually until the posthibernation period. These findings suggest that differences of lectin binding in the thumb pad may be related to functional activities and, thus, seasonal adaptations. Moreover, the presence of specific lectin‐binding cells in the breeding glands indicated that they consisted of heterogeneous secretory cell composition or that the cells were at different secretory stages. J. Morphol. 275:76–86, 2014. © 2013 Wiley Periodicals, Inc.     

Evolution and phylogeny of gonad morphology in bony fishes

Gonad morphology at the gross anatomical or histological levelshas long been studied by fisheries biologists to identify annualreproductive cycles and length of breeding season, among othergoals. Comparative surveys across vertebrate taxa have not beendetailed enough, however, to describe fully the differencesand similarities among gonads of bony fishes and other vertebrates,and to use gonad morphology in phylogenetic systematic analyses.An emerging constant among vertebrates is the presence of agerminal epithelium composed of somatic and germ cells in bothmales and females. In females, the germinal epithelium linesthe ovarian lamellae. In males, arrangement of the germinalepithelium into compartments varies among osteichthyans: basaltaxa have an anastomosing tubular testis, whereas derived taxahave a lobular testis. The lobular testis is proposed as a synapomorphyof the Neoteleostei. The annual reproductive cycle is hypothesizedto be the source of morphological variation among testis types.Elongation of germinal compartments during early maturationmay result in a transition from anastomosing tubular to lobulartestes. In all male atherinomorphs surveyed, spermatogonia arerestricted to the distal termini of lobules rather than beingdistributed along the lobule; there is an epithelioid arrangementof Sertoli and germ cells rather than a germinal epithelium.Arrest of the maturation-regression phases is hypothesized tolead to formation of the atherinomorph testis. Atherinomorphsalso have a distinctive egg with fluid, rather than granular,yolk. Variation among germinal epithelia is interpreted in adeveloping phylogenetic framework to understand evolution ofgonad morphology and to propose gonad characters for phylogeneticanalyses.     

45T-1, an established cell line with characteristics of Sertoli cells, forms organized aggregates in vitro after exposure to tumor necrosis factor alpha.

In the testis TNF is produced by germinal cells. The putative role of tumor necrosis factor alpha (TNF) in development and differentiation was investigated in 45T-1 mouse cell cultures, a cell line with characteristic markers of Sertoli cells, established from transgenic mouse families expressing the polyoma large T antigen in their testes. Exposure to TNF elicited a gradual assembly of the cells of the monolayer into highly organized spheroids. The first morphological sign of the changes was detected one week after TNF treatment by anti-desmin immunostaining which showed the formation of foci in the culture consisting of several hundred cells connected by an increasing number of cell contacts. Between days 10-20 the cells formed large ovoid or vermiform aggregates covered by several layers of flat, elongated cells. These cells extended septae into the inner mass of the spheroids consisting of loosely arranged, large polygonal or palisadic cells. The spheroids were surrounded by radially arranged elongated cells covered by small blebs. TNF treatment upregulated laminin expression in 45T-1 cell cultures, which is known to induce formation of cord-like structures by Sertoli cells in vitro. Coculturing 45T-1 cells with immortalized germinal cells or TNF-producing HeLa cells also lead to the formation of spheroids. These observations suggest that TNF production of germinal cells might contribute to the organization/differentiation of Sertoli cells.     

Involvement of the gonadal germinal epithelium during sex reversal and seasonal testicular cycling in the protogynous swamp eel,Synbranchus marmoratus Bloch 1795 (Teleostei,Synbranchidae)

The swamp eel, Synbranchus marmoratus, is a protogynous, diandric species. During sex reversal, the ovarian germinal epithelium, which forms follicles containing an oocyte and encompassing follicle cells during the female portion of the life cycle, produces numerous invaginations, or acini, into the ovarian stroma. Within the acini, the gonia that formerly produced oocytes become spermatogonia, enter meiosis, and produce sperm. The acini are bounded by the basement membrane of the germinal epithelium. Epithelial cells of the female germinal epithelium, which formerly became follicle (granulosa) cells, now become Sertoli cells in the developing testis. Subsequently, lobules and testicular ducts form. The swamp eel testis has a lobular germinal compartment in both primary and secondary males, although the germinal compartment in testes of secondary males resides within the former ovarian lamellae. The germinal compartment, supported by a basement membrane, is composed of Sertoli and germ cells that give rise to sperm. Histological and immunohistochemical techniques were used to describe the five reproductive classes that were observed to occur during the annual reproductive cycle: regressed, early maturation, mid-maturation, late maturation, and regression. These classes are differentiated by the presence of continuous or discontinuous germinal epithelia and by the types of germ cells present. Synbranchus marmoratus has a permanent germinal epithelium. Differences between the germinal compartment of the testes of primary and secondary males were not observed.