多肠目薄背涡虫生殖系统结构及一氧化氮合酶组化研究

运用常规组织学方法和NADPH-d组织化学方法,研究了薄背涡虫 Notoplana humilis 生殖系统的组织结构和一氧化氮合酶的分布.其雄性生殖系统包括精巢、储精囊、阴茎、雄性生殖孔,精巢壁由一薄层薄膜组成,每个精巢内都含有不同发育时期的雄性生殖细胞,且精子发育无明显同步性;储精囊呈螺旋状排列在雄性生殖孔附近,囊壁由单层扁平上皮组成;阴茎为粗大的球形,外壁由柱状上皮细胞和数层肌细胞组成.雌性生殖系统包括输卵管、生殖腔、雌性生殖孔和受精囊,但不形成集中的卵巢和卵黄腺.雌雄生殖孔、生殖腔、受精囊、阴茎等部位呈NADPH-d强阳性反应.     

中国涡虫一新纪录科肠口涡虫属一新纪录种格氏肠口涡虫(原卵黄目, 柱口科)

原卵黄目(Prolecithophora)柱口科(Cylindrostomidae)肠口涡虫属(Enterostomula)为海栖涡虫,全球记录3种,在中国未见报道。作者在广东省深圳市深圳湾海边(22°31'N,113°56'E)采集到一种海栖涡虫。本文对该种涡虫进行了较为详细的比较研究,鉴定为柱口科肠口涡虫属格氏肠口涡虫(E.graffi de Beauchamp,1913),为中国新纪录科新纪录属一新纪录种。该种涡虫多数个体背部具2条黑色横纹,部分个体背部花纹有明显变化。眼点2对。雌雄同体,精巢、卵巢各1个。受精囊位于卵巢与子宫之间;子宫位于体末端,后接一微小的外阴道。本文发现:①该涡虫的尾部腹面具有一个交配囊及一根雌性生殖管,交配囊是一个由肌肉层组成的袋状囊,同时与子宫、雌性生殖管相连;②雌性生殖管外围有明显的生殖腺包裹;③雌性生殖管孔、阴茎孔与口孔一起通往体外;④此种涡虫喜分泌黏液将微小杂质粘结呈半球状窝,虫体隐藏于窝内。     

长蛸生殖系统的形态学与组织学观察

运用解剖学和组织学方法对长蛸(Octopus variabilis)生殖系统的形态结构进行了研究.结果表明,长蛸雌雄异体并异形,雄性右侧第三腕茎化.长蛸雌性生殖系统由一个卵巢、成对的输卵管及输卵管腺组成.卵巢壁上发出一条线状具分支的生殖索,米粒状的滤泡以卵柄连接至生殖索上.每个滤泡是由单层滤泡细胞围绕着一个卵母细胞构成.输卵管形成丰富的纵行褶皱,黏膜上皮具有纤毛.输卵管腺含有两种类型腺细胞.雄性生殖系统包括精巢、输精管前段、储精囊、摄护腺、盲囊、输精管后段和精荚囊.精巢内部被结缔组织分隔成许多精小叶,精原细胞由小叶壁中的生殖上皮产生,并向小叶腔中逐步分化成精子.输精管前段、盲囊和摄护腺所分泌的黏液物质共同参与精荚的形成.储精囊和输精管后段形成较多的纵行褶皱,输精管后段上皮游离面的纤毛可运输生殖细胞.精荚囊的作用则是贮存精荚,囊壁中的平滑肌利于长蛸交配时精荚的排出.     

平疣桑椹石磺生殖系统结构及精子储存场所

雌雄同体贝类精子的储存和利用规律一直是国内外贝类生物学研究的难点之一,本文利用活体解剖、显微观察、组织切片和扫描电镜技术,综合研究了平疣桑椹石磺(Platevindex mortoni)的生殖系统及精子储存场所。结果显示,其生殖系统包括生殖器本部、雌性生殖部分和雄性生殖部分。生殖器本部由两性腺、两性输送管、蛋白腺、黏液腺、支囊组成;雌性生殖部分包括输卵管、受精囊、阴道,位于身体中后方体腔内;雄性生殖部分包括输精管、刺激器、阴茎、阴茎鞘和阴茎牵引肌,位于身体前端右侧体腔内;其阴茎有阴茎鞘,阴茎表面布满倒刺。平疣桑椹石磺阴茎为直线状,无雄性附属腺。未交配的性成熟个体支囊内充满细长精子,受精囊内无精子;而交配后充当雌性个体的支囊内均为细长的自体精子,受精囊内有大量活力较强的粗短精子,其支囊为自体精子的存储场所,而受精囊为异体精子的存储场所。其精子储运情况为:两性腺内精子成熟后暂存于支囊,交配时通过输精管运输至阴茎,由阴茎输送精子至对方的阴道,异体精子进入受精囊内存储待用。     

瘤背石磺的生殖系统结构特点及生殖周期研究

采用活体性腺观察和组织切片方法对瘤背石磺的生殖系统及生殖周期进行了周年的研究.结果表明,瘤背石磺的生殖系统由三部分组成:两性腺、雄性生殖管道及附属腺体、雌性生殖管道及附属腺体.雄性生殖管道由输精管、阴茎、刺激器组成,附性腺为主要的附属腺体;雌性生殖管道由输卵管、受精囊、阴道组成,附属腺体主要为卵黄腺和蛋白腺.瘤背石磺的生殖周期具有典型的腹足类动物的特征,性腺发育周期可以分为:休止期、增殖期、生长期、成熟期、排放期5个时期,生殖周期为1年,成熟和排放期在6月上旬到9月上旬,繁殖高峰期在7月上旬至8月上旬,10月后进入休止期.本文还讨论了瘤背石磺在繁殖过程中精卵的输送、受精等过程及相关的交配器和附属腺体的功能.     

长足大竹象生殖系统的形态解剖研究

解剖研究了长足大竹象雌雄虫牛殖系统的构造.该虫的雌性生殖系统包括一对卵巢、一对侧输卵管、中输卵管、交配囊、受精囊、生殖腔、产卵器;雄性生殖系统由一对睾九、一对输精管、一对附腺、射精管和交配器组成.     

粉尘螨生殖系统超微结构的透射电镜观察

本研究主要采用透射电镜观察粉尘螨Dermatophagoides farinae (Hughes)生殖系统超微结构。粉尘螨雄性生殖系统是由精巢、 输 精管、 附腺、 射精管、 交配器官及附属交配器官组成。精巢内可同时有精子发育各阶段的细胞。精子无核膜、 核染色质聚集成束、 线 粒体缺乏典型的嵴、 胞质内有平行排列的电子致密薄片等为其特征性结构。雌性生殖系统由交合囊、 交合囊管、 储精囊、 囊导管、 卵 巢、 输卵管、 子宫及产卵管构成。卵巢内可见含多个细胞核的中央细胞, 其周为卵母细胞等生殖细胞。该研究丰富了对粉尘螨生殖系统 结构的认识。     

双雄性交配器官大口涡虫的首次发现

大口涡虫属所有物种均为雌雄同体,具一套交配器官。作者于2015年在广东省的两处淡水环境,首次发现2个具有双雄性交配器官的大口涡虫(Macrostomum sp.)标本。通过对活体、整装片、连续组织切片的显微镜观测,发现2只标本的两套交配器官呈左右排列;每套交配器官具备完整的假储精囊、储精囊、颗粒囊与交配刺;假储精囊与储精囊内具有精子;雄孔分别为1个与2个;交配刺的结构与中国已经记录的物种都不相同。本研究对其做了较为详细的描述,并初步探讨了大口涡虫多交配器官发生的原因。     

瘤背石磺的形态、习性和生殖行为

2003年5月～2004年5月研究了上海崇明瘤背石磺(Onchidium struma)的形态特征、生活习性,分别对其消化、呼吸、循环、排泄、生殖、神经等六大系统进行了阐述。结果表明,瘤背石磺生活在潮间带高潮区滩涂的芦苇丛里,摄食泥滩上的有机质和单胞藻类;雌雄同体、异体交配、卵生。生殖系统包括生殖器和雌、雄交接器三部分：生殖器由两性腺、卵黄腺和蛋白腺组成;雄性交接器由输精管、附性腺、阴茎、刺激器等组成,雄性生殖孔位于右侧第一触角中部;雌性交接器由输卵管、受精囊、阴道等组成,雌性生殖孔位于肛门右侧约5．0mm处的腹足与外套膜的交界处。本文并详细描述了石磺的交配行为。     

瘤背石磺产卵前后生殖系统的组织学变化

瘤背石磺(Onchidium struma)是雌雄同体、异体交配的腹足纲贝类,其生殖系统较为复杂,通过解剖学和组织切片技术对成体瘤背石磺的生殖系统及产卵前后的组织学变化进行了系统的研究.结果表明:(1)雄性生殖系统主要南阴茎囊、阴茎、雄性附性腺、两性腺(早期主要产生精于)和储精囊等部分组成,而雌性生殖系统则由两性腺(后期主要产生卵子)、生殖细胞输送管、蛋白腺、黏液腺、受精囊和阴道等组成;(2)雄性生殖系统的组织学结构在产卵前后变化较小,但两性腺、卵蛋白腺和黏液腺的组织学在产卵前后变化显著;(3)产卵后的两性腺由于成熟卵子的排放,整体结构松散,部分腺泡中有少量未排出的成熟卵细胞和卵黄合成早期的卵母细胞;(4)产卵前的卵蛋白腺中含有许多强嗜碱性的小颗粒(组织学结构类似于卵鞘中的胚胎外周蛋白),产卵后腺体中的颗粒相对较大,且呈嗜酸性;(5)产卵前的黏液腺中存在嗜碱性区、嗜酸性区和混杂区三种区域,但是产卵前黏液腺以嗜酸性细胞为主,而产卵后的黏液腺中以嗜碱性细胞区域为主,且分泌管道中有一些嗜碱性物质.由此可见,卵蛋白腺的主要功能是分泌卵蛋白包裹受精卵形成卵外周蛋白层,而黏液腺则在产卵过程中分泌黏液物质形成卵鞘结构及链状的卵带.     

Detection and changes in levels of testosterone during spermatogenesis in the freshwater planarian Bdellocephala brunnea

It was reported recently that vertebrate-type steroids exist and control reproduction in several groups of invertebrates, including molluscs. Sexually reproductive freshwater planarians of the species Bdellocephala brunnea have a limited breeding season in their natural habitat. This phenomenon suggests that some endogenous reproductive hormones might play a role in vivo. However, to date, sex steroids such as androgen, estrogen, and progesterone have not been found in planarians. The goal of the present study was to determine whether androgen is present in sexual planarians such as B. brunnea. The presence of testosterone was detected by high-pressure liquid chromatography and, in sexually reproductive individuals in which no seminal vesicles were visible, the level of testosterone was about twice than that in individuals with visible seminal vesicles. An enzyme-linked immunosorbent assay revealed that the levels of testosterone during terminal spermatogenesis were three times higher than during the spermatocyte-building phase. Our results indicate that sexually reproductive freshwater planarians such as B. brunnea might have vertebrate-type steroids and show variation in testosterone levels during spermatogenesis.     

Effects of 17β-estradiol and bisphenol A on the formation of reproductive organs in planarians

Planarians have a remarkable capacity for regeneration after ablation, and they reproduce asexually by fission. However, some planarians can also reproduce and maintain their sexual organs. During the regenerative process, their existing sexual organs degenerate and new ones develop. However, little is known about hormonal regulation during the development of reproductive organs in planarians. In this study, we investigated the effects of 17β-estradiol (a steroid) and bisphenol A (an endocrine disrupter) on the formation of sexual organs in the hermaphroditic planarian Dugesia ryukyuensis. Under control conditions, all worm tissues regenerated into sexual planarians with sexual organs within 4 weeks after ablation. However, in the presence of bisphenol A or 17β-estradiol, although they apparently regenerated into sexual planarians, the yolk glands, which are one of the female sexual organs, failed to regenerate even 7 weeks after ablation. These data suggest that planarians have a steroid hormone system, which plays a key role in the formation and maturation of sexual organs.     

Some details on the morphological structure of planarian musculature identified by fluorescent and confocal laser-scanning microscopy

The details of the morphological organization of the body musculature in the planarians Girardia tigrina and Polycelis tenuis were investigated by histochemical staining of actin filaments with fluorescently labeled fluorescent. The whole mount preparations and frozen tissue sections of planarians were analyzed by fluorescent and confocal laser scanning microscopy. The results indicate that the muscle system is well differentiated in both planarian species and is represented by the somatic musculature of the body wall, the musculature of the digestive tract, and the musculature of the reproductive system organs in P. tenuis, which reproduces sexually. The differences and similarities between the two species in the morphological characters of the musculature, which are the size and density of myofibrils in different muscle layers, were described. The results present the basis for further studies on the regulation of muscle function in planarians.     

Genome-wide analyses reveal a role for peptide hormones in planarian germline development

Bioactive peptides (i.e., neuropeptides or peptide hormones) represent the largest class of cell-cell signaling molecules in metazoans and are potent regulators of neural and physiological function. In vertebrates, peptide hormones play an integral role in endocrine signaling between the brain and the gonads that controls reproductive development, yet few of these molecules have been shown to influence reproductive development in invertebrates. Here, we define a role for peptide hormones in controlling reproductive physiology of the model flatworm, the planarian Schmidtea mediterranea. Based on our observation that defective neuropeptide processing results in defects in reproductive system development, we employed peptidomic and functional genomic approaches to characterize the planarian peptide hormone complement, identifying 51 prohormone genes and validating 142 peptides biochemically. Comprehensive in situ hybridization analyses of prohormone gene expression revealed the unanticipated complexity of the flatworm nervous system and identified a prohormone specifically expressed in the nervous system of sexually reproducing planarians. We show that this member of the neuropeptide Y superfamily is required for the maintenance of mature reproductive organs and differentiated germ cells in the testes. Additionally, comparative analyses of our biochemically validated prohormones with the genomes of the parasitic flatworms Schistosoma mansoni and Schistosoma japonicum identified new schistosome prohormones and validated half of all predicted peptide-encoding genes in these parasites. These studies describe the peptide hormone complement of a flatworm on a genome-wide scale and reveal a previously uncharacterized role for peptide hormones in flatworm reproduction. Furthermore, they suggest new opportunities for using planarians as free-living models for understanding the reproductive biology of flatworm parasites.     

A low percent ethanol method for immobilizing planarians

Planarians have recently become a popular model system for the study of adult stem cells, regeneration and polarity. The system is attractive for both undergraduate and graduate research labs, since planarian colonies are low cost and easy to maintain. Also in situ hybridization, immunofluorescence and RNA-interference (RNAi) gene knockdown techniques have been developed for planarian studies. However, imaging of live worms (particularly at high magnifications) is difficult because animals are strongly photophobic; they quickly move away from light sources and out of frame. The current methods available to inhibit movement in planarians include RNAi injection and exposure to cold temperatures. The former is labor and time intensive, while the latter precludes the use of many fluorescent reporter dyes. Here, we report a simple, inexpensive and reversible method to immobilize planarians for live imaging. Our data show that a short 1 hour treatment with 3% ethanol (EtOH) is sufficient to inhibit both the fine and gross movements of Schmidtea mediterranea planarians, of the typical size used (4-6 mm), with full recovery of movement within 3-4 hours. Importantly, EtOH treatment did not interfere with regeneration, even after repeated exposure, nor lyse epithelial cells (as assayed by H&E staining). We demonstrate that a short exposure to a low concentration of EtOH is a quick and effective method of immobilizing planarians, one that is easily adaptable to planarians of all sizes and will increase the accessibility of live imaging assays to planarian researchers.     

TCAV-1, a monoclonal antibody specific to epithelial pharyngeal cells in the planarian Dugesia (Girardia) tigrina. Application to pattern formation of the pharynx during regeneration

During regeneration in planarians, anterior (head and prepharyngeal) and posterior (postpharyngeal and tail) fragments rebuild one of the most peculiar structures of planarians: the pharynx and the pharynx cavity. Previous studies (see Brønsted, 1969, for a general review, and Asai, 1990, 1991, for anterior regeneration) have shown that within postpharyngeal pieces both structures appear in the old stump from clusters of undifferentiated cells. However, the lineage and differentiation of their elements (inner and outer epithelial cells, muscle layers, gland cells, nerve rings) and the overall pattern of growth and differentiation is not clear.     

A histological study of the accessory reproductive organs of female Loligo forbesi (Cephalopoda: Loliginidae)

In Loligo forbesi Steenstrup, the female reproductive system consists of the ovary and accessory reproductive organs which include the oviducal gland, the nidamental gland, the accessory nidamental gland and seminal receptacle. Histological studies were made on the accessory reproductive organs of female L. forbesi. The various changes observed during maturation are described and the functional significance discussed. The secretions produced by the oviducal gland and nidamental gland apparently form the egg coats. The seminal receptacle serves to store spermatozoa after mating. The function of the accessory nidamental gland is unknown.     

淡水涡虫染色体的制备方法

以涡虫的再生组织为材料,用秋水仙素处理长有再生组织的涡虫片段,并通过改善各种实验条件,得到高清晰度的染色体图谱。结果表明,本方法简便易行,制成的染色体分散好,形态清晰,适用于对涡虫染色体的进一步研究。