Cyclic Adenosine Monophosphate: A Central Player in Gamete Development and Fertilization,and Possible Target for Infertility Therapies

Human infertility, of both male and female origin, is often caused by the deficient response of the testis and the ovary to hormonal stimuli that govern sperm and oocyte development and fertilization. The effects of hormones and other extracellular ligands involved in these events are often mediated by G-protein-coupled receptors that employ cyclic adenosine monophosphate (cAMP) as the principal second messenger transducing the receptor-generated signal to downstream elements. This opinion article summarizes the actions of cAMP in sperm and oocyte development and fertilization, leading to therapeutic actions targeting cAMP metabolism to alleviate human male and female infertility.     

Knockdown of the Halloween Genes spook,shadow and shade Influences Oocyte Development,Egg Shape,Oviposition and Hatching in the Desert Locust

Ecdysteroids are widely investigated for their role during the molting cascade in insects; however, they are also involved in the development of the female reproductive system. Ecdysteroids are synthesized from cholesterol, which is further converted via a series of enzymatic steps into the main molting hormone, 20-hydoxyecdysone. Most of these biosynthetic conversion steps involve the activity of cytochrome P450 (CYP) hydroxylases, which are encoded by the Halloween genes. Three of these genes, spook (spo), phantom (phm) and shade (shd), were previously characterized in the desert locust, Schistocerca gregaria. Based on recent sequencing data, we have now identified the sequences of disembodied (dib) and shadow (sad), for which we also analyzed spatiotemporal expression profiles using qRT-PCR. Furthermore, we investigated the possible role(s) of five different Halloween genes in the oogenesis process by means of RNA interference mediated knockdown experiments. Our results showed that depleting the expression of SchgrSpo, SchgrSad and SchgrShd had a significant impact on oocyte development, oviposition and hatching of the eggs. Moreover, the shape of the growing oocytes, as well as the deposited eggs, was very drastically altered by the experimental treatments. Consequently, it can be proposed that these three enzymes play an important role in oogenesis.     

美洲鲥鱼卵巢发育规律和性类固醇激素变化研究

采用组织学、形态学及电化学等技术方法,研究了人工养殖条件下美洲鲥鱼雌性亲鱼卵巢发育和性类固醇激素的周期变化规律。结果表明,美洲鲥鱼属于非同步分批产卵类型;根据卵巢外部形状大小及色泽等的不同,可将美洲鲥鱼卵巢发育过程分为6个时期;根据卵母细胞的形态结构、卵黄物质的积累和滤泡细胞的变化,将卵子发生划分为6个时相;卵巢周期变化过程中,肝脏指数(HSI)和肥满度(CF)变化不显著,性腺指数(GSI)与血浆中雌二醇(E2)水平表现出与性腺发育协同一致的变化规律。     

大鳍鳠卵发生中溶酶体超微结构变化与卵膜发生

大鳍鳠卵细胞发生中的溶酶体结构变化活跃,Ⅰ期卵原细胞胞质内可见线粒体、高尔基体和少量呈球状的初级溶酶体。Ⅱ期卵母细胞胞质内可见多个球状溶酶体,溶酶体内的水解酶蛋白呈结晶态,多紧贴线粒体部分分布。卵母细胞质膜绉缩,胞质边缘可见大量球状和线状溶酶体分布。Ⅲ期卵母细胞内侧出现绒毛状卵膜结构,外侧出现“放射带”,球状的初级溶酶体结构消失,绒毛状卵膜与放射带之间可见大量线状溶酶体结构。推测球状溶酶体和线粒体间接参与了卵母细胞质膜的绉缩位移;而线状溶酶体则可能直接参与了绒毛状卵膜和“放射带”结构的形成。     

Elemental composition of pyroantimonate precipitates analysed by electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS) in vitellogenic ovarian follicles of Drosophila

Ca²⁺ was precipitated with potassium antimonate in vitellogenic follicles of the fruit fly Drosophila melanogaster and the distribution of the precipitates formed was studied by electron microscopy. The microvilli of the oolemma in mid- and late vitellogenic follicles were lined with precipitates. The chemical composition of the precipitates was analysed by electron spectroscopic imaging (ESI). The images produced by inelastically scattered electrons at specific ionization edges were compared, and the nonspecific background signals were subtracted by an image processing system. The presence of Ca²⁺, antimony and oxygen in the precipitates formed could be demonstrated. The elemental composition of the precipitates and of yolk spheres was also analysed by electron energy-loss spectroscopy (EELS). With respect to the precipitates, signals at the calcium L_2,3-edge, the oxygen K-edge and the antimony M_4,5-edge were recorded without deconvolution and background subtraction. The yolk spheres, which were free of precipitates, gave the characteristic signal of the nitrogen K-edge. The applied techniques combine good ultrastructural resolution with the possibility of analysing the elemental composition of histochemical reaction products and cellular structures.     

泥螺卵子发生过程中高尔基复合体的变化

应用透射电子显微镜研究了泥螺（Bullacta exarata）卵子发生过程中高尔基复合体的数量和形态结构特点.结果表明,在泥螺卵子发生的四个时期即卵原细胞期、卵黄发生早期、卵黄发生中期和卵黄发生后期,卵母细胞内高尔基复合体经历了一系列的变化.卵原细胞内无典型的高尔基复合体,但具高尔基囊泡;卵黄发生早期,高尔基复合体发达,膜囊数达10多层;卵黄发生中期,高尔基复合体分泌活动加快,产生大量的大小囊泡,囊泡内充满颗粒状物质,最后演变成卵黄颗粒.随着卵母细胞的发育,卵黄颗粒的数量和直径逐渐增加;卵黄发生后期,高尔基复合体开始解体,数量减少.此外,对高尔基复合体在卵子形成过程中的功能、去向及行为变化等问题进行了讨论.     

Cytochemistry of fat body trophocytes and ovaries of workers and queens of Melipona quadrifasciata anthidioides (Hymenoptera: Apidae: Meliponini) during vitellogenesis

ABSTRACT The fat body (FB) of insects is where yolk proteins are synthesized. Therefore, relationships between the FB and oogenesis were studied in nurse workers, virgins, and physogastric queens of Melipona quadrifasciata anthidioides, a stingless bee in which the workers produce and lay eggs while provisioning brood cells. The relationships between FB and oogenesis, as well as the routes of materials from hemolymph to the oocytes, were studied through the cytochemical detection of lipids by osmium imidazole (OI), carbohydrates by ruthenium red (RR) and basic proteins by ammoniacal silver (AS). The results show differences in the presence of the studied materials in FB trophocytes and ovary of the classes of females studied and oogenesis phases. Material that tested positive for the treatments was detected among the classes of individuals studied in both, trophocytes and oocytes, and in the route of those materials from hemolymph to the oocytes. The differences found among the individual classes indicate relationships with the nutrition and adaptation to the parsimonious use of nutrients in the metabolism of reproduction. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.     

A Soluble Pyrophosphatase Is Essential to Oogenesis and Is Required for Polyphosphate Metabolism in the Red Flour Beetle (Tribolium castaneum)

Polyphosphates have been found in all cell types examined to date and play diverse roles depending on the cell type. In eukaryotic organisms, polyphosphates have been mainly investigated in mammalian cells with few studies on insects. Some studies have demonstrated that a pyrophosphatase regulates polyphosphate metabolism, and most of them were performed on trypanosomatids. Here, we investigated the effects of sPPase gene knocked down in oogenesis and polyphosphate metabolism in the red flour beetle (Tribolium castaneum) A single sPPase gene was identified in insect genome and is maternally provided at the mRNA level and not restricted to any embryonic or extraembryonic region during embryogenesis. After injection of Tc-sPPase dsRNA, female survival was reduced to 15% of the control (dsNeo RNA), and egg laying was completely impaired. The morphological analysis by nuclear DAPI staining of the ovarioles in Tc-sPPase dsRNA-injected females showed that the ovariole number is diminished, degenerated oocytes can be observed, and germarium is reduced. The polyphosphate level was increased in cytoplasmic and nuclear fractions in Tc-sPPase RNAi; Concomitantly, the exopolyphosphatase activity decreased in both fractions. Altogether, these data suggest a role for sPPase in the regulation on polyphosphate metabolism in insects and provide evidence that Tc-sPPase is essential to oogenesis.