A missense mutation in the Capza3 gene and disruption of F-actin organization in spermatids of repro32 infertile male mice

Males homozygous for the repro32 ENU-induced mutation produced by the Reproductive Genomics program at The Jackson Laboratory are infertile, have low epididymal sperm concentrations, and produce sperm with abnormally shaped heads and poor motility. The purpose of the present study was to identify the mutated gene in repro32 mice and to define the structural and functional changes causing infertility and the aberrant sperm phenotype. In repro32/repro32 mice, we discovered a failure to shed excess cytoplasm and disorganization of the middle piece of the flagellum at spermiation, resulting in the outer dense fibers being wrapped around the sperm head within a bag of cytoplasm. Using a candidate-gene approach, a mutation was identified in the spermatid-specific “capping protein (actin filament) muscle Z-line, alpha 3” gene (Capza3). CAPZA3 protein localization was altered in spermatids concurrent with altered localization of a unique CAPZB variant isoform and disruption of the filamentous actin (F-actin) network. These observations strongly suggest the missense mutation in Capza3 is responsible for the mutant phenotype of repro32/repro32 sperm and regulation of F-actin dynamics by a spermatogenic cell-specific CAPZ heterodimer is essential for removal of the cytoplasm and maintenance of midpiece integrity during spermiation in the mouse.     

海月水母精巢发育及排精过程的观察

采用实验生态学及显微观察的方法研究了海月水母(Aurelia sp.)的精巢发育及其排精过程,并对其精子活力进行了测定。结果表明:在水温20~22℃的条件下,海月水母碟状体经过40 d生长,达到伞径(7.50±0.71)cm、体重(28.70±6.60)g时,精巢出现并生长发育;经过60 d生长,达到伞径(11.77±0.51)cm、体重(83.54±10.36)g时,精巢发育成熟并开始排精;生长90 d后,精巢开始出现退化,当生长110 d时,精巢退化完全。在精巢发育过程中,其宽度和长度分别伴随海月水母伞径的增长而增宽和伸长,并出现折叠现象。海月水母的排精路线为:精子先粘附于精子细丝上,从精巢排出,继而经过胃循环沟、胃口腕沟,最后由口腕基沟排出体外。在水温22℃、盐度30、p H 8.0的条件下,海月水母精子活力随时间延长而降低,其快速运动时间和寿命分别为4 h 30 min和10 h。本研究结果显示,在适宜的环境条件下,海月水母精巢发育迅速,排精路线与过程相对简单,其精子活力强、寿命长,这种高效的生殖策略为其暴发奠定了基础,这或许也是海月水母能在地球上存活年代久远的原因之一。     

排卵,排精前后文昌鱼体内GnRH的研究

利用放射免疫分析法测定了排卵、排精前后青岛文昌鱼体内促性腺激素释放激素（ＧｎＲＨ）的含量变化,并通过高效液相色谱比较了雌、雄文昌鱼性腺及除性腺外体部ＧｎＲＨ的种类和含量的异同。结果表明：１）生殖过程中雌、雄文昌鱼体内ＧｎＲＨ含量的变化规律不同;雌性文昌鱼体内ＧｎＲＨ总含量在排卵时有所下降,排后１２小时政策最为明显,以后逐渐上升到排前水平;雄性文昌鱼仅在排精时有所下降,２小时后即稳定在排水平。２）文     

Effects of age, weight, hormones, and hibernation on breeding success in boreal toads (Bufo boreas boreas)

The goals of this study were to test the effects of exogenous hormones and hibernation on breeding behavior and gamete release by boreal toads (Bufo boreas boreas). Each year, a subset of 77 toads was hibernated and then paired with hibernated or nonhibernated mates and treated with luteinizing hormone releasing hormone analogue (LHRHa), human chorionic gonadotropin (hCG), or left untreated. Amplexus and egg and sperm production were recorded. At 1 yr of age, only 19% of pairs exhibited amplexus, and no sperm or eggs were produced. At 2 and 3 yr of age, most male toads treated with LHRHa exhibited amplexus (56.9% and 100%, respectively). Among 2-yr-old males, amplexus was more prevalent (P < 0.05) in those that were hibernated than in those that were nonhibernated (54.0% and 33.3%, respectively), but most males in each group (93.3% and 75%, respectively) produced sperm in response to LHRHa treatment. Only one 2-yr-old and two 3-yr-old females produced eggs. At 4 yr of age, eight females produced eggs, but two died from egg retention. More nonhibernated than hibernated females developed eggs (7 of 10 vs. 1 of 10, P < 0.05). Mean (±SD) weight of female toads producing eggs (58.9 ± 11.9 g) was greater (P < 0.05) than that of nonproducing females (43.6 ± 7.0 g). Similarly, four of seven nonhibernated females (58.8 ± 8.3 g) produced eggs at 5 yr of age. All eggs were produced by females treated once with LHRHa. Number of eggs per female varied (141 to 3307), and development to tadpoles was low (0 to 36.5%), although tadpoles did become toadlets. In conclusion, male and female boreal toads matured at 2 and 4 yr of age, respectively, and heavier females were more likely to produce eggs. To enhance breeding success, males should be hibernated and treated with LHRHa. In contrast, female productivity was enhanced by improving their body condition instead of subjecting them to hibernation prior to LHRHa treatment.     

Sex steroids in plasma of lake whitefish Coregonus clupeaformis during spawning in Lake Erie

Lake whitefish, Coregonus clupeaformis, were collected from the Western basin of Lake Erie during spawning. Free and conjugated (sulfated and glucuronidated) steroids including testosterone (T), 11-ketotestosterone (11-kT), estradiol-17beta (E2) and 17,20beta-dihydroxy-4-pregnen-3-one (17,20betaP) were measured in the plasma by radioimmunoassay. In males, the progression of spermiation was characterized by a significant decrease in plasma free steroids, whereas the levels of conjugated steroids remained similar and low, except for sulfated and glucuronidated testosterone. Plasma sex steroids did not correlate with the density or the motility of the spermatozoa. In females, the concentration of plasma T was significantly higher in preovulating than in ovulating females. The levels of E2 and 17,20betaP in ovulating lake whitefish exhibited large variations ranging from below detection limit to 0.9 ng ml(-1) and from 0.2 to 13 ng ml(-1), respectively. Analysis of conjugated steroids revealed high levels of glucuronidated and sulfated 17,20betaP and glucuronidated T in females ovulating in December. However, no significant differences in the proportion of the conjugated steroids were observed.     

The ultrastructure of the Sertoli cell of the vervet monkey, Chlorocebus aethiops

The ultrastructure of the Sertoli cell of the vervet monkey was studied using both scanning and transmission electron microscopic techniques. SEM micrographs revealed perforated sleeve-like processes which encased mature elongated spermatids which are ready for spermiation. TEM micrographs showed a large Sertoli cell nucleus characterized by many lobes (4–5) and consisting of a homogenous nucleoplasm and a distinctive nucleolus. The nucleus occupies a significant portion of the basal region of the cell. The distribution of chromatin clearly shows high activity of these cells. Lipid droplets and free ribosomes are also found scattered throughout the cytoplasm. Well-developed Golgi apparatus is found in the basal region of the cell. There is phagocytic activity in the Sertoli cells as revealed by the presence of numerous phagosomes. Numerous mitochondria with well-developed tubular cristae are found on the basal side of the nucleus, whereas few mitochondria are located on the apical side of the nucleus. Distinct desmosomes are located between cells. A well-developed smooth endoplasmic reticulum and granular endoplasmic reticulum are frequently found in the cytoplasm of the Sertoli cells. The results of this investigation showed that Sertoli cells of the vervet monkey are almost similar to those of humans and show many similarities with other mammalian species.     

Effects of photoperiod and temperature on testicular development in male ayu,Plecoglossus altivelis

Synopsis Growth, gonadosomatic index and plasma steroid profiles in male ayu,Plecoglossus altivelis, cultured under short/long photoperiods and cool/warm temperatures were determined. Juvenile males were assigned to each of four different photoperiod/temperature regimes (16 L/18°C, 16 L/24°C, 8 L/18°C and 8 L/24°C) at random. Fish were killed and examined bi-weekly over the following 16 weeks. Mean body weight in the 16 L/18°C treated fish was the highest among four treated groups. No significant differences between body weights of the 16 L/24°C, 8 L/18°C and 8 L/24°C treated groups were observed. Ayu in the 8 L/18°C treated group had the highest values of gonadosomatic index, plasma testosterone (T) and 17-hydroxyprogesterone (17-OH P). No significant differences of plasma E₂ were observed among the treated groups. In the 8L/18°C and 8L/24°C groups, peak levels of 17-OH P occurred after 12 and 14 weeks of treatment, respectively. No peak levels of plasma T and 17-OH P were observed in 16 L/18°C or 16 L/24°C treated ayu. Spermiation occurred only in ayu with 8 L/18°C treatment. The data suggest that testicular development in ayu is temperature and photoperiod dependent: short photoperiod and cool temperature favour gonadal development.     

Relationship between steroidogenesis and spermiation in Rana catesbeiana and Leptodactylus ocellatus

The present study employs an in vitro system to analyse the role of steroid hormones in hCG-induced spermiation in two species of anuran amphibian: Rana catesbeiana and Leptodactylus ocellatus. In vitro spermiation was induced with 10 IU hCG and the effect of different steroid-biosynthesis inhibitors was analysed. Cyanoketone (10⁻⁵ M), an inhibitor of 3-oxo-4-ene steroid biosynthesis, did not block hCG-inducing activity even when biosynthesis of androgen was significantly reduced. These results clearly showed that, in both species, spermiation-inducing action of hCG does not depend on the biosynthesis of 3-oxo-4-ene steroids. Moreover, when combined inhibitors, aminoglutethimide (10⁻⁵ M) plus cyanoketone (10⁻⁵ M), were employed, spermiation evoked by hCG was not modified while hCG-induced androgen secretion significantly decreased. Additionally, none of the steroids used, progesterone, 17, 20α-dihydroxy-4-pregnen-3-one, testosterone and 5α-dihydrotestosterone, were able to induce spermiation in the absence of hCG, confirming that steroids are not involved in that process. In conclusion, as previously described in Bufo arenarum, in L. ocellatus and R. catesbeiana hCG-induced spermiation does not depend on steroid biosynthesis.     

Inhibition of spermiation in the syrian hamster using dibutyryl cyclic-AMP

Summary Young adult male Syrian hamsters were given intraperitoneal injections of 50 mg dibutyryl cyclic AMP twice daily for a period of three days. On the fourth day the animals were sacrificed and their testes were processed for light and electron microscopy. The results indicate that the mature spermatozoa were retained within the seminiferous epithelium after the stage in the seminiferous cycle in which spermiation normally occurs. The unreleased spermatozoa were ultrastructurally normal. Typical Sertoli-spermatid junctional specializations remained associated with the retained spermatozoa. These findings indicate that normally spermiation is initiated by the disappearance of the junctional specializations. In addition, the present results demonstrate that spermiation can be controlled.Supported in part by NIH Grant RR05654 and by NIH Grant P 30 HD 10202 (Morphology Core)The skillful technical assistance of Pam Duke is gratefully acknowledged