Spermiogenesis and sperm ultrastructure in ten species of Loricariidae (Siluriformes, Teleostei)

The current study describes the ultrastructural characteristics of spermatogenesis, spermiogenesis, and spermatozoa in specimens of siluriform taxa Neoplecostominae, Hypoptopomatinae, Otothyrinae, Loricariinae, and Hypostominae. Our data show that the characteristics of spermatogenesis and spermiogenesis and spermatozoa ultrastructure of Neoplecostominae are more common to Hypoptopomatinae and Otothyrinae than to Loricariinae and Hypostominae. Furthermore, Loricariinae and Hypostominae have more characteristics in common than with any other group of Loricariidae. These data reinforce the phylogenetic hypotheses of relationships among the subfamilies of Loricariidae. Considering the available data in Loricarioidei, Loricariidae presents ultrastructural characteristics of spermatogenesis and spermiogenesis that are also observed in Astroblepidae, its sister group. However, the most of the characteristics of spermatozoa ultrastructure found in Astroblepidae are also observed in Scoloplacidae, the sister group of a clade composed of Astroblepidae and Loricariidae.     

Gamma-ray exposure accelerates spermatogenesis of medaka fish,Oryzias latipes

To examine the spermatogenesis (and spermiogenesis) cell population kinetics after gamma-irradiation, the frequency and fate of BrdU-labeled pre-meiotic spermatogenic cells (spermatogonia and pre-leptotene spermatocytes) and spermatogonial stem cells (SSCs) of the medaka fish (Oryzias latipes) were examined immunohistochemically and by BrdU-labeling. After 4.75 Gy of gamma-irradiation, a statistically significant decrease in the frequency of BrdU-labeled cells was detected in the SSCs, but not in pre-meiotic spermatogenic cells. The time necessary for differentiation of surviving pre-meiotic spermatogenic cells without delay of germ cell development was shortened. More than 90% of surviving pre-meiotic spermatogenic cells differentiated into haploid cells within 5 days after irradiation, followed by a temporal spermatozoa exhaust in the testis. Next, spermatogenesis began in the surviving SSCs. However, the outcome was abnormal spermatozoa, indicating that accelerated maturation process led to morphological abnormalities. Moreover, 35% of the morphologically normal spermatozoa were dead at day 6. Based on these results, we suggest a reset system; after irradiation most surviving spermatogenic cells, except for the SSCs, are prematurely eliminated from the testis by spermatogenesis (and spermiogenesis) acceleration, and subsequent spermatogenesis begins with the surviving SSCs, a possible safeguard against male germ cell mutagenesis.     

Follicle-stimulating hormone induces spermatogenesis mediated by androgen production in Japanese eel, Anguilla japonica

Follicle-stimulating hormone (FSH) plays important roles in spermatogenesis. However, the biologic activity of FSH can vary in different vertebrate classes, and the definitive function of FSH has not been established. In this study, we investigated the functions of FSH on spermatogenesis using an in vitro culture system for Japanese eel testis. The eel Fsh receptor was expressed in testis tissue during the whole process of spermatogenesis, mainly by Leydig cells that produce steroid hormones and by Sertoli cells surrounding type A spermatogonia and early type B spermatogonia. In an in vitro organ culture, recombinant eel Fsh (r-eFsh) induced complete spermatogenesis from the proliferation of spermatogonia to spermiogenesis during 36 days of culture; also, spermatozoa were observed in the testicular fragments. Spermatogenesis induced by r-eFsh was inhibited by trilostane, a specific inhibitor of 3beta-hydroxysteroid dehydrogenase. However, trilostane did not inhibit spermatogenesis induced by 11-ketotestosterone. These results clearly show that the main function of FSH in eel is to induce spermatogenesis via stimulating androgen production.     

Sperm of Doradidae (Teleostei: Siluriformes)

Spermatic characteristics were studied in 10 species representing several distinct groups within the catfish family Doradidae. Interestingly, different types of spermatogenesis, spermiogenesis and spermatozoa are correlated with intrafamilial groups previously proposed for Doradidae. Semi-cystic spermatogenesis, modified Type III spermiogenesis, and biflagellate sperm appear to be unique within Doradidae to the subfamily Astrodoradinae. Other doradid species have sperm with a single flagellum, cystic spermatogenesis, and spermiogenesis of Type I (Pterodoras granulosus, Rhinodoras dorbignyi), Type I modified (Oxydoras kneri), or Type III (Trachydoras paraguayensis). Doradids have an external mode of fertilization, and share a few spermatic characteristics, such as cystic spermatogenesis, Type I spermiogenesis and uniflagellate sperm, with its sister group Auchenipteridae, a family exhibiting sperm modifications associated with insemination and internal fertilization. Semi-cystic spermatogenesis and biflagellate spermatozoa are also found in Aspredinidae, and corroborate recent proposals that Aspredinidae and Doradoidea (Doradidae + Auchenipteridae) are sister groups and that Astrodoradinae occupies a basal position within Doradidae. The co-occurrence in various catfish families of semi-cystic spermatogenesis and either biflagellate spermatozoa (Aspredinidae, Cetopsidae, Doradidae, Malapturidae, Nematogenyidae) or uniflagellate sperm with two axonemes (Ariidae) reinforces the suggestion that such characteristics are correlated. Semi-cystic spermatogenesis and biflagellate sperm may represent ancestral conditions for Loricarioidei and Siluroidei of Siluriformes as they occur in putatively basal members of each suborder, Nematogenyidae and Cetopsidae, respectively. However, if semi-cystic spermatogenesis and biflagellate sperm are ancestral for Siluriformes, cystic spermatogenesis and uniflagellate sperm have arisen independently in multiple lineages including Diplomystidae, sister group to Siluroidei.     

北方山溪鲵精子发生不同阶段的显微与超微结构

用光镜和电镜观察了北方山溪鲵(Batrachuperus tibetanus)精子发生过程中各种类型生精细胞的显微与超微结构变化。结果显示,北方山溪鲵在4~8月时处于精子发生期,精子形成在7~8月。成熟精子的结构具有小鲵科精子的一些共同特征,如顶体前端呈三叶草状,尾部由轴纤维、波动膜、轴丝及轴丝旁纤维构成,轴纤维粗大呈圆柱形,尾部无线粒体等。比较分析认为,在两栖类的系统发育中,轴纤维、波动膜和轴丝旁纤维的消失为近裔性状。     

Inhibition of Spermiogenesis of Eupyrene Spermatozoa by Elevated,Sublethal Temperatures During Pupal-Adult Development of the Bertha Armyworm,Mamestra con figurata Wlk. (Lepidoptera,Noctuidae)

Summary

Eupyrene spermiogenesis and spermatozoa production in Mamestra configurata Walker were inhibited by temperatures >24° C during a “critical period” of about 7 days during the first half of post-diapause pupal-adult development. The critical period coincided with the appearance of early-stage eupyrene spermatids. If temperatures >24° C occurred during the critical period, eupyrene spermatozoa production was reduced, the result of irreversible autolysis of the early- and mid-stage eupyrene spermatids. Each day of exposure to 25 or 27.5° C during the critical period reduced the number of eupyrene cysts with spermatozoa on average by 12 to 14%. There were no observable effects on eupyrene spermiogenesis and spermatozoa production of exposing post-diapausing pupae to 25 or 27.5° C before or after the critical period. Apyrene spermatogenesis was not affected by 25 or 27.5° C.

Exposure of pupae to 25°or 27.5° C during diapause had no apparent effect on eupyrene spermiogenesis and spermatozoa production. Meiosis usually was not detected among the eupyrene cysts with lalje primary spermatocytes until after diapause, indicating that meiosis is suppressed during diapause. By providing a meiotic block, diapause governs the time of onset of the temperature-sensitive critical period of spermiogenesis. In nature, male pupae of M. configurata in diapause are protected from sterilizing temperatures in the soil during August and post-diapause developing pupae pass through the critical period of spermiogenesis in spring (April to June) when soil temperatures throughout the range of M. configurata in Canada are well below 25° C. The distribution of M. configurata in the northern region of the temperate zone may in part be attributable to the sensitivity of spermatogenesis to relatively mild temperatures.     

Abnormal spermatogenesis at low temperatures in the Japanese red-bellied newt,Cynops pyrrhogaster: possible biological significance of the cessation of spermatocytogenesis

In newt testis, spermatocytes never appear during winter, because secondary spermatogonia die by apoptosis just before meiosis. In the current study, we examined the effect of low temperatures on spermatogenesis. Incubation of newts at low temperatures (8, 12, 15 degrees C) induced defects in spermatogenesis in a temperature-dependent manner. At 8 degrees C, multinucleated giant cells (MGCs) were observed in spermatocytes and spermatogenesis never proceeded beyond meiosis. Although spermatocytes completed meiotic divisions at 12 degrees C, severe cell death was observed in the spermatids. At 15 degrees C both normal and abnormal spermiogenesis were observed. Under these conditions, impaired meiotic synapsis/recombination and down-regulation of the expression of the DMC1 protein, which play pivotal roles in meiotic pairing in eukaryotes, were also observed. Furthermore, to examine the quality of the sperm produced at low temperature for supporting development, artificial insemination was performed. The eggs inseminated with spermatozoa derived from newts kept at 15 degrees C demonstrated a restricted developmental capacity, even though these spermatozoa had an equal capacity for carrying out fertilization to those kept at 22 degrees C. These results suggest that meiosis at low temperatures cause the production of abnormal spermatozoa. Conservation and the significance of this phenomenon in poikilothermic vertebrates living in the temperate zones are also discussed.     

秀丽线虫精子发生和精子受精的研究进展

秀丽线虫精子发生过程包括减数分裂和精子活化两个阶段, 通过早期特异基因的表达和后期蛋白分子的翻译后修饰, 精原细胞发育成为具有运动能力的精子。其受精阶段包括精子运动、精子竞争、精卵信号通讯以及精卵融合等过程。通过突变体筛选目前已经获得了一些影响精子发生或受精的突变体, 并且对其中一些突变体进行了基因克隆和功能分析的研究。这些研究不仅对于阐明精子发生和受精的机理具有重大的理论意义, 而且对男性不育的治疗和男性无毒避孕药物的研发可能提供重要的依据。文章阐述了目前在线虫精子发生和精子受精两个方面的研究进展。     

The testicular cycle of Gambusia affinis holbrooki (Teleostei: Poeciliidae)

Fifteen male mosquito fish ( Gambusia affinis holbrooki ) were collected in 1989 on the 15th of each month to perform a quantitative histologic study of the annual testicular cycle including a calculation of the gonadosomatic index, testicular volume, and the total volume per testis occupied by each germ cell type. The cycle comprises two periods: spermatogenesis and quiescence. The spermatogenic period begins in April with the development of primary spermatogonia into secondary spermatogonia, spermatocytes and round spermatids. In May, the first spermatogenic wave is completed and the testicular volume begins to increase up to June when the maximum testicular volume and gonadosomatic index are reached. Germ cell proliferation with successive spermatogenetic waves continues until August. In September germ cell proliferation ceases and neither secondary spermatogonia nor spermatocytes are observed. However, spermiogenesis continues until October. In November, spermiogenesis has stopped and the testis enters the quiescent period up to April. During this period only primary spermatogonia and spermatozoa are present in the testis. In addition, a few spermatids whose spermiogenesis was arrested in November are observed. Testicular release of spermatozoa is continuous during the entire spermatogenesis period. The spermatozoa formed at the end of this period (September-October) remain in the testis during the quiescent period and are released at the beginning of the next spermatogenesis period in April. Developed Leydig cells appear all year long in the testicular interstitium, mainly around both efferent ducts and the testicular tubule sections showing S₄ spermatids.     

Ultrastructure of Spermatogenesis in the Testis of Paragonimus heterotremus

Lung fluke, Paragonimus heterotremus, is a flatworm causing pulmonary paragonimiasis in cats, dogs, and humans in Southeast Asia. We examined the ultrastructure of the testis of adult P. heterotremus with special attention to spermatogenesis and spermiogenesis using scanning and transmission electron microscopy. The full sequence of spermatogenesis and spermiogenesis, from the capsular basal lamina to the luminal surface, was demonstrated. The sequence comprises spermatogonia, spermatocytes with obvious nuclear synaptonemal complexes, spermatids, and eventual spermatozoa. Moreover, full steps of spermatid differentiation were shown which consisted of 1) early stage, 2) differentiation stage representing the flagella, intercentriolar body, basal body, striated rootlets, and electron dense nucleus of thread-like lamellar configuration, and 3) growing spermatid flagella. Detailed ultrastructure of 2 different types of spermatozoa was also shown in this study.     

Spermatogenesis and spermiogenesis in Microcotyle sp. (Microcotylidae, Monogenea)

Ultrastructural observations of spermatogenesis and spermiogenesis in Microcotyle sp. a microcotylid monogenean parasite from the gills of Hypostomus sp., are described. The spermatogonia were irregularly shaped, forming a peripheral layer of cells; spermatocytes were larger than spermatogonia and a nuclear synaptonemal complex was observed; young spermatids were joined by a central cytophore forming rosettes. Spermiogenesis was characterized by the outgrowth of a cytoplasmic protuberance, the zone of differentiation, containing the basal bodies, separated by an intercentriolar body, from which flagella grow out and fuse posteriorly with the median process. Cross sections of the anterior and the middle regions of spermatozoa revealed nuclei, mitochondria, peripheral microtubules, and paired axonemes each with a 9+1 pattern.     

Occurrence of biflagellate spermatozoa in Cetopsidae, Aspredinidae, and Nematogenyidae (Teleostei: Ostariophysi: Siluriformes)

In the present study spermiogenesis was investigated in Cetopsis coecutiens (Cetopsidae), and Bunocephalus amazonicus (Aspredinidae), while spermatozoa ultrastructure was investigated in C. coecutiens, B. amazonicus, and Nematogenys inermis (Nematogenyidae). Aspredinidae and Cetopsidae share a spermatogenesis of the semicystic type, and a particular type of spermiogenesis process not reported in any fish group. In the three species analyzed, spermatozoa are biflagellate with flagella having the classical axoneme formulae (9 + 2). The analysis of thirteen characters showed the presence of eight characters shared by Cetopsidae and Aspredinidae, and six characters shared by Cetopsidae and Nematogenyidae, which may suggest that these three families may be more related than actually hypothesized, comprising a very primitive siluriform lineage originated after Diplomystidae.     

Continuous gametogenesis in the neotropical freshwater teleost, Bryconops affinis (Pisces:Characidae)

The gametogenesis of Bryconops affinis was studied by light, transmission and scanning electron microscopy. The spermatogenesis is semi-cystic and spermatids are released into the lumen of seminiferous tubules, where spermiogenesis is completed. Spermatozoa have an ovoid head, a rudimentary middle piece with a small number of mitochondria and long flagellum (primitive spermatozoa). The Sertoli and Leydig cells show secretory activity during spermatogenesis. By the end of this phenomenon, the Sertoli cells phagocytize the residual spermatozoa, while the Leydig cells show involuted characteristics. With regard to the oogenesis process, the oocyte development was divided into four stages based on the cytological characteristics of the oocyte and its surrounding layers. Ultrastructural analysis revealed that the zona pellucida is formed during the previtellogenic stage. Specializations associated to the outer layer of the zona pellucida may be related to the egg's adherence to the substrata.     

Initiation and kinetic profiles of spermatogenesis in the frog, Runa esculenta (Amphibia)

Spermatogenesis in Rana esculenta is initiated during metamorphic climax and mature spermatozoa are present in froglets 45 days old. Cytological analysis of cell populations shows that some of the primary spermatogonia may lie dormant for brief intervals of time. Timing analysis of the process of spermatogenesis, in adults and in developing Frog larvae maintained at approximately 18°C, was investigated by different methods. The results are remarkably similar. Although perfect synchrony of the developing cells within a single germinal cyst is not the rule, a uniform rate of advancement of germinal cysts of the same stage of development, in most of the seminiferous tubules of a testis is evident. The duration of the secondary spermatogonial divisions is five to six days, involving five or six mitotic cycles, each cycle lasting approximately 24 h. The premeiotic S-phase, and phases of leptotene, zygotene, pachytene, diplotene+secondary spermatocytes, and spermiogenesis each have a duration, respectively, of six, two, six, twelve, two and seven days. The duration of spermatogenesis, from a "committed" primary spermatogonium to the formation of spermatozoa is 41 days.     

Temporal synthesis of mitochondrial proteins from mouse epididymal spermatozoa

Mitochondria from ejaculated bovine spermatozoa contain a group of polypeptides ranging in molecular weights from 13,000 to 35,000 not found in other bovine or murine testicular mitochondria [Hecht and Bradley, 1981]. These proteins are present in the mitochondria isolated from both epididymal and ejaculated spermatozoa. To establish when during epididymal transport, spermiogenesis, and/or meiosis these proteins are synthesized, the synthesis intervals for the mitochondrial proteins from cauda epididymal spermatozoa were established following intratesticular injection of (³⁵S)methionine. Mice were killed every third day over a 33-day period and cauda epididymal spermatozoa were fractionated into mitochondrial and head components. Radioactivity in each fraction was monitored by liquid scintillation counting. Maximal incorporation was observed during spermiogenesis, although substantial amounts of protein were synthesized during meiosis. Analysis of the mitochondrial polypeptides by gel electrophoresis revealed that many polypeptides such as the cysteine-rich structural protein of the mitochondrial capsule were synthesized over prolonged intervals of spermiogenesis and meiosis rather than in a brief specific time period. These results suggest that spermatozoal mitochondria are produced by a sequential substitution of new proteins into the differentiating mitochondria rather than the abrupt appearance of a new class of mitochondria during spermatogenesis.     

Estrogen,through estrogen receptor 1, regulates histone modifications and chromatin remodeling during spermatogenesis in adult rats

Estrogen receptors (ESR1 and ESR2) play crucial roles in various processes during spermatogenesis. To elucidate individual roles of ESRs in male fertility, we developed in vivo selective ESR agonist administration models. Adult male rats treated with ESR1 and ESR2 agonist for 60 days show spermatogenic defects leading to reduced sperm counts and fertility. While studying epigenetic changes in the male germ line that could have affected fertility, we earlier observed a decrease in DNA methylation and its machinery upon ESR2 agonist treatment. Here, we explored the effects on histone modifications, which could contribute to decreased male fertility upon ESR agonist administration. ESR1 agonist treatment affected testicular levels of histone modifications associated with active and repressed chromatin states, along with heterochromatin marks. This was concomitant with deregulation of corresponding histone modifying enzymes in the testis. In addition, there was increased retention of histones along with protamine deficiency in the caudal spermatozoa after ESR1 agonist treatment. This could be due to the observed decrease in several chromatin remodeling proteins implicated in mediating histone-to-protamine exchange during spermiogenesis. The activating and repressing histone marks in spermatozoa, which play a critical role in early embryo development, were deregulated after both the ESR agonist treatments. Together, these epigenetic defects in the male germ line could affect the spermatozoa quality and lead to the observed decrease in fertility. Our results thus highlight the importance of ESRs in regulating different epigenetic processes during spermatogenesis, which are crucial for male fertility.     

Ultrastructure of the semicystic spermatogenesis in the South American freshwater characid Hemigrammus marginatus (Teleostei,Characiformes)

Semicystic, a rare type of spermatogenesis, was detected in the characid Hemigrammus marginatus and characterized by cysts hatching during the spermatid phase and maturation of the spermatozoa being completed at the lumen of the anastomosed seminiferous tubules. Primary spermatogonia, or type A, are distributed along the entire length of the seminiferous tubules, in an unrestricted spermatogonial pattern. H. marginatus spermiogenesis is included in type I, mainly characterized by presence of nucleus rotation. During this process, a vesicle resembling the acrosomal vesicle is visualized at the anterior region close to the nucleus of the early spermatids, however this structure did not remain in the spermatozoa. In H. marginatus, the spermatozoon is uniflagellated, primitive, type I aquasperm, with a rounded head, a short midpiece and a long flagellum with the axoneme in a 9 + 2 microtubules arrangement and no lateral fins. Residual spermatozoa are reabsorbed by Sertoli cells. Unusual biflagellate spermatozoa with three long cytoplasmatic projections originating in the midpiece are rarely observed and have not been registered in other characiforms. Ultrastructural characteristics of the spermatogenesis and spermatozoa observed in the present work provide important subsidies to systematic and phylogeny studies of Characidae fishes included in Incertae sedis groups, such as H. marginatus.