Reproductive biology of Philodryas patagoniensis (Snakes: Dipsadidae) in south Brazil: male reproductive cycle

The male reproductive cycle of Philodryas patagoniensis in south Brazil was described through morpho‐anatomical and histological analysis of individuals deposited in zoological collections. Spermatogenesis occurred during late autumn–winter (June–September) and spermiogenesis occurred in spring–summer (October–March). The volume of the testes was smaller (quiescent) in winter, while the tubular diameter and the epithelial height of the seminiferous tubule were larger in summer (January–March). The ductus deferens presented spermatozoa all over the year and had no seasonal variation in diameter. The length of the kidney was larger in winter–spring (July–December), although the tubular diameter and epithelium height of the sexual segment of the kidney (SSK) were larger only in winter (July–September). Total testicular regression was observed in late autumn (May), simultaneously with the peak in SSK. Therefore, at the individual level, males exhibit a discontinuous cyclical reproduction. Considering the population level, the reproductive cycle is seasonal semisynchronous, with most of the individuals showing a reproductive peak in spring–summer (October–March). Here, we present evidence to support the importance of the microscopic approach to reproductive cycle studies. Finally, we discuss the intrinsic and extrinsic factors influencing P. patagoniensis reproductive patterns.     

Sperm storage in males of the snake Crotalus durissus terrificus (Crotalinae: Viperidae) in southeastern Brazil

Seasonal variations in spermatozoa numbers and in sperm motility along the vas deferens in Crotalus durissus terrificus from southeastern Brazil were analyzed. Our data demonstrate storage and motility of the spermatozoa along the vas deferens throughout the year. This is characteristic of a postnuptial reproductive cycle, usually found in snakes living in temperate climates. We describe similarities in reproductive cycle patterns found in the tropical nonhibernator C. durissus terrificus and in hibernator snakes from temperate zones. Our results show that in C. durissus terrificus, a significant difference in spermatozoa counts occurs between winter and summer. Higher numbers of spermatozoa in summer and autumn, due to intense spermiogenesis, coincides with the mating season in autumn. These data indicate that after spermiogenesis in summer, the males combine the peak of sperm storage to the period females are attractive. Mating, however, is not linked to ovulation, and the sperm is stored in the females during winter until fertilization occurs in spring. In the males, after mating, spermatozoon counts low. In spring, they gradually increase, turning again the highest in summer and autumn. During spermiogenesis in the convoluted vas deferens, spermatozoa gain motility, enhancing their performance along their way towards the distal portion.     

Ultrastructure of sperm storage and male genital ducts in a male holocephalan, the elephant fish, Callorhynchus milii.

In chondrichthyes, the process of spermatogenesis produces a spermatocyst composed of Sertoli cells and their cohort of associated spermatozoa linearly arrayed and embedded in the apical end of the Sertoli cell. The extratesticular ducts consist of paired epididymis, ductus deferens, isthmus, and seminal vesicles. In transit through the ducts, spermatozoa undergo modification by secretions of the extratesticular ducts and associated glands, i.e., Leydig gland. In mature animals, the anterior portion of the mesonephros is specialized as the Leydig gland that connects to both the epididymis and ductus deferens and elaborates seminal fluid and matrix that contribute to the spermatophore or spermatozeugmata, depending on the species. Leydig gland epithelium is simple columnar with secretory and ciliated cells. Secretory cells have periodic acid-Schiff positive (PAS+) apical secretory granules. In the holocephalan elephant fish, Callorhynchus milii, sperm and Sertoli cell fragments enter the first major extratesticular duct, the epididymis. In the epididymis, spermatozoa are initially present as individual sperm but soon begin to laterally associate so that they are aligned head-to-head. The epididymis is a highly convoluted tubule with a small bore lumen and an epithelium consisting of scant ciliated and relatively more secretory cells. Secretory activity of both the Leydig gland and epididymis contribute to the nascent spermatophores, which begin as gel-like aggregations of secretory product in which sperm are embedded. Fully formed spermatophores occur in the ductus. The simple columnar epithelium has both ciliated and secretory cells. The spermatophore is regionalized into a PAS+ and Alcian-blue-positive (AB+) cortex and a distinctively PAS+, and less AB+ medulla. Laterally aligned sperm occupy the medulla and are surrounded by a clear zone separate from the spermatophore matrix. Grossly, the seminal vesicles are characterized by spiral partitions of the epithelium that project into the lumen, much like a spiral staircase. Each partition is staggered with respect to adjacent partitions while the aperture is eccentric. The generally nonsecretory epithelium of the seminal vesicle is simple columnar with both microvillar and ciliated cells.     

Distribution of acid glycosidases in the male genital tract of the pheasant

The distribution of the activity of seven acid glycosidases in the reproductive organs of the pheasant (Phasianus colchicus) was studied. The study was carried out on seven mature birds at the age of 11 months during the reproductive season (May). Significant (p<0.01) differences in acid glycosidase activity dependent on enzyme origin were observed. Generally, the highest activity of acid glycosidases was found in the epididymis, intermediate in the ductus deferens and the lowest in the testes. Exceptionally, alpha-mannosidase had the highest activity in the ductus deferens. Anion-exchange chromatography elution profiles of most enzymes from the tested reproductive organs were similar, however evident differences were observed for beta-mannosidase forms.     

Microanatomy of the testes and testicular ducts of the butterfly lizard,Leiolepis ocellata Peters, 1971 (Reptilia: Squamata: Agamidae) during the active reproductive period

The microanatomy of the testes and testicular ducts (rete testis, ductuli efferentes, ductus epididymis and ductus deferens) of Leiolepis ocellata (Agamidae) was investigated using light microscopy including histochemistry. Each testis contains seminiferous tubules and interstitial tissues. The former house spermatogenic cells (spermatogonia A & B, preleptotene, primary and secondary spermatocytes, spermatids (steps 1–8) and spermatozoa) and Sertoli cells, while the latter comprise peritubular and intersitial tissues. The rete testis is an anastomosing duct, having intratesticular and extratesticular portions. The proximal region of ductuli efferentes has wider outer ductal and luminal diameters than those of the distal region. The convoluted ductus epididymis is subdivided into four regions (initial segment, caput, corpus and cauda), based on the ductal diameter, epithelium characteristics and cell components. The ductus deferens has the greatest diameter and is divided into the ductal and ampulla ductus deferens. The ductal portion is subdivided into the proximal and distal regions, based on the epithelium types and ductal diameters. The ampulla ductus deferens is a fibromuscular tube, having numerous mucosal folds projecting into the lumen. Spermiophagy is detectable in the ductus epididymis and ductus deferens. The present results contribute to improved fundamental knowledge on the microanatomy of the reptilian reproductive system.     

Growth related changes in the content of heparin and 5-hydroxytryptamine of mast cells

Summary Fine structural changes of testicular interstitial cells of Leydig and secretory cells of seminal vesicles were studied in golden hamsters under different functional states of the pineal gland. Experiments were performed in the reproductive season (summer months). In the hamsters blinded for 8 weeks the testes and the seminal vesicles were markedly atrophic, and the Leydig cells and the secretory cells of seminal vesicles were extremely involuted. By contrast, both types of cells in the pinealectomized or superior cervical ganglionectomized hamsters exhibited cytological features suggestive of an enhanced secretory activity. This study shows that functional activity of Leydig cells as well as secretory cells of seminal vesicles in the hamster may be depressed or augmented by stimulating or inhibiting the pineal antigonadal function, respectively, without performing hypophysectomy or hormonal administration.Dedicated to Professor Shu Yeh on the occasion of his 70th birthday. This study was supported in part by a grant from the National Science Council, the Republic of China     

Reproductive biology of Philodryas patagoniensis (Snakes: Dipsadidae) in south Brazil: Female reproductive cycle

In this study, we describe the female reproductive cycle of Philodryas patagoniensis in south Brazil, which was described through morpho‐anatomical and histological analyses. The peak of secondary vitellogenesis occurred during winter–spring (July–December), ovulation in spring (October–December), mating and fertilization in spring–summer (October–February), oviposition in spring–autumn (October–May) and births from late spring to autumn (December–July). The diameter of vitellogenic follicles/eggs was larger in winter–spring than in other seasons. The diameter of the shell glands was also larger in winter–spring. In spite of the clear reproductive peak, gonads only showed reduced activity in the autumn. Therefore, at the individual level, females have a discontinuous cyclical reproduction; in the populational level, the reproductive cycle is seasonal semisynchronous. We support the hypothesis that P. patagoniensis have the ability to produce multiple clutches with long‐term stored sperm. Sexual dimorphism in body size was evident, and females are significantly larger and heavier than males. Larger females were able to produce follicles and eggs in larger amount and size. The maternal body size was positively related to the reproductive effort and fecundity. To conclude, we deliberated about the proximal and distal causes that influence the reproductive traits and patterns of P. patagoniensis.     

Ultrastructure and histochemistry of the male reproductive system of the genus Callinectes Stimpson, 1860 (Brachyura: Portunidae)

We described the ultrastructure and histochemistry of the reproductive system of five Callinectes species, and evaluate the seasonal variation in weight of the reproductive system and hepatopancreas by comparing annual changes of somatic indices. The somatic indices changed little throughout the year. In Callinectes, spermatogenesis occurs inside the lobular testes and, within each lobule, the cells are at the same developmental stage. Spermatogenesis and spermiogenesis follow the same development pattern in all Callinectes studied. Mature spermatozoa are released into the seminiferous ducts through the collecting ducts. Cells of the vas deferens are secretory as evidenced by rough endoplasmic reticulum, Golgi complex, and secretory vesicles that produce the seminal fluid. The anterior vas deferens shows two portions: proximal and distal. In proximal portion (AVDp), spermatozoa are clustered and embedded in an electron-dense, basophilic glycoproteinaceous secretion Type I. In the distal portion (AVDd), the spermatophore wall is formed by incorporation of a less electron-dense glycoproteinaceous secretion Type II. The secretion Type I change to an acid polysaccharide-rich matrix that separates the spermatophores from each other. The median vas deferens (MVD) stores the spermatophores and produces the granular glycoproteinaceous seminal fluid. The posterior vas deferens (PVD) has few spermatophores. Its epithelium has many mitochondria and the PVD seminal fluid changes into a liquid and homogeneous glycoprotein. Many outpocketings in the PVD and MVD help to increase the fluid production. Overall, the reproductive pattern of Callinectes is similar to other species that produce sperm plugs. The secretions of AVD, MVD, and PVD are responsible for the polymerization that forms the solid, waxy plug in the seminal receptacle. The traits identified here are common to all Portunidae species studied so far.     

Seasonal Ultrastructural Changes of Leydig Cells in Lacerta muralis

We have studied the ultrastructure of the testis and interstitial tissue of lizard Lacerta muralis sacrificed in spring, summer and autumn, with special emphasis on the morphological changes of Leydig cells. From the autumn to the end of spring, Leydig cells showed a large smooth endoplasmic reticulum and mitochondria with tubular cristae. These features correlate with the synthesis and secretion of androgens. At the end of spring and after mating, the amount of smooth endoplasmic reticulum decreased and the mitochondria showed laminar cristae. Both features are typical of a latent period, during which time secretion of androgens remains inactive until the end of summer. The possible role of other cell organelles, such as Golgi complex and lysosome-like electron dense bodies, during both secreting and resting periods, is also discussed. Finally, we consider the relationship between seasons, secretory activity of Leydig cells and the spermatogenic cycle.     

Reproductive pattern in the male Angolan free-tailed bat, Tadarida (Mops) condylura (Microchiroptera: Molossidae) in the Eastern Transvaal, South Africa

The reproductive pattern in male Angolan free-tailed bats Tadarida (Mops) condylura was studied at two localities in the Eastern Transvaal during the period 1989-1991. Males displayed a nine-month breeding season extending from June to February. Following testicular recrudescence in early June, spermatogenic activity displayed two periods of peak activity in August/early September and November/early December. Although testes became involuted in March, baseline spermatogenic activity was maintained and tests never regressed to prepubertal conditions. Leydig cell morphology closely followed spermatogenic activity, with the diameter of Leydig cell nuclei displaying a concomitant seasonal pattern.     

Male reproductive seasonality of the Snake-eyed Lizard,Ophisops elegans Ménétriés, 1832, from Lebanon (Reptilia: Lacertidae)

We studied the male reproductive cycle in a population of Ophisops elegans from Mount Sannine, Lebanon, by histological analysis. Testicular histology showed active spermatogenesis in spring, followed by a testicular regression at the end of summer and a subsequent recrudescence in autumn. Monthly variations in the epididymis, the ductus deferens and the sexual segment of the kidney were in synchrony with the testicular cycle. They were hypertrophied as spermatogenetic activity increased and atrophied as spermatogenetic activity decreased. Males of O. elegans showed a vernal type of spermatogenesis with a close relationship between the evolution of the seminiferous tubules and the secondary sexual characters.     

Reproductive morphology of the male Tuatara,Sphenodon punctatus

Over the past decade, studies on reproductive morphology in the Squamata (snakes and lizards) have expanded tremendously. With the accumulation of these studies and revisions of the terminology based on structural similarities and differences, it is imperative to review the work on tuataras to determine whether the structural organization fits the revised terminology of vertebrates. We investigated the morphology of the male reproductive system in the Tuatara, Sphenodon punctatus (Rhynchocephalia), the sister taxon to the Squamata. Previous studies on the Tuatara used a nomenclature for the testicular ducts different from the current terminology for amniotes. The reproductive system in the Tuatara is consistent with reports in the Squamata. Two rete testis tubules exit the testis within a connective tissue sheath similar to that shown in other squamate species and the protherian Echidna. Each rete testis divides into multiple ductuli efferentes that fuse with the epididymis. The epididymis transitions into the ductus deferens where the sperm become more concentrated into spherical bundles. The ductus deferens enters the cloacal urodeum separately from the ureter. An ampulla ureter or ampulla urogenital papilla was not observed, which differs from previous studies of lepidosaurians. Furthermore, a sexual segment of the kidney (SSK) was not observed, consistent with previous studies on the Tuatara.     

Nutrition,sex and season contribute to variation in fat and glycerol levels in the long‐lived moth Caloptilia fraxinella

The ash leaf cone roller Caloptilia fraxinella Ely (Lepidoptera: Gracillariidae) is an invasive leaf‐mining moth pest of horticultural ash Fraxinus spp. in the Canadian Prairie Provinces. Caloptilia fraxinella overwinter as adults in reproductive diapause and mating occurs after overwintering in the spring. The effect of a carbohydrate food source on fat and glycerol reserves throughout the long adult life stage of this moth is investigated. Insects collected as pupae are given access to either water or sugar water upon adult eclosion. Moths held under the different feeding regimes are sampled before (summer and autumn) and after overwintering in the spring. Analysis of either glycerol or lipid content is conducted for male and female moths from each collection period. Both moth weight and glycerol concentration are affected by moth sex, food regime and season of collection. Although female moths are heavier than males, a higher glycerol concentration occurs in males. Moths fed sugar are heavier and have a higher glycerol concentration than water‐fed moths late in reproductive diapause and after overwintering. Moths collected in the spring after overwintering are lighter and have a lower glycerol content than moths collected before winter. There is a significant influence of feeding regime and season on moth body lipid content, with sugar‐fed moths having more fat than water‐fed moths; however, this difference is smaller in the summer than the autumn or spring. An initial understanding of the overwintering biology and diapause of this pest is provided in the present study.     

Reproduction of the blacktip sawtail catshark,Galeus sauteri,in the waters off northeastern Taiwan

The reproductive biology of blacktip sawtail catsharks,Galeus sauteri, in northeastern Taiwan waters was investigated. Male catsharks possessed paired testes producing spermatozoa, which were then stored in the epididymides all year round. No spermatophores were observed in the lower ductus deferens. Only the right ovary was functional in females, oogonia being formed in the ovarian cortex and then developing into mature ova. Ova exceeding 17 mm in diameter were ovulated. Pregnant individuals contained one eggcase only, in each side of the uterus. No hatched embryos were observed in such eggcases. The size at which 50% of blacktip sawtail catshark specimens were mature was 410–420 mm and 350–360 mm for females and males, respectively. The species does not have a well-defined reproductive season.     

Occurrence of ampulla in the ductus deferens of the Indian garden lizard Calotes versicolor daudin

During the breeding season, the terminal end of the ductus deferens of Calotes versicolor appears swollen and is comparable to the ampulla of the mammalian ductus deferens. Its anatomy was studied from paraffin sections. It differentiates along its length into five zones. The first has thick smooth muscle and pesudostratified epithelium; the second has luminal trabeculae with an epithelium showing evidence of secretory activity; the third has the epithelial mucosa abutting against the smooth muscle in the form of pocketlike indentations; the fourth has crypts between epithelial folds; and the fifth zone is a sphincter. The anatomy of this ampullary region is indicative of secretory as well as spermatophagous roles. It undergoes seasonal change and appears to be androgen-dependent. © 1995 Wiley-Liss, Inc.     

Peptidase activities in the semen from the ductus deferens and uterus of the neotropical rattlesnake Crotalus durissus terrificus

To understand the role of peptidases in seminal physiology of Crotalus durissus terrificus, intra- and inter-seasonal activity levels of acid (APA), basic (APB), puromycin-sensitive (APN-PS) and puromycin-insensitive neutral (APN-PI), cystyl (CAP), dipeptidyl-IV (DPPIV), type-1 pyroglutamyl (PAP-I) and prolyl-imino (PIP) aminopeptidases as well as prolyl endopeptidase (POP) were evaluated in soluble (SF) and/or membrane-bound (MF) fractions of semen collected from the ductus deferens of the male reproductive tract and from the posterior portion of the uterus. Seminal APB, PIP and POP were detected in SF, while other peptidases were detected in SF and MF. Only the convoluted posterior uterus in winter and autumn had semen. Relative to other examined peptidases, in general, APN-PI, APN-PS and APB activities were predominant in the semen from the uterus and throughout the year in the semen from the ductus deferens, suggesting their great relevance in the seminal physiology of C. d. terrificus. The levels of peptidase activities in the ductus deferens semen varied seasonally and were different from those of semen in the uterus, suggesting that their modulatory actions on susceptible peptides are integrated to the male reproductive cycle events and spermatozoa viability of this snake.     

Histological structure of the male reproductive organs and spermatogenesis in a copulating sculpin, <Emphasis Type="Italic">Radulinopsis taranetzi</Emphasis> (Scorpaeniformes: Cottidae)

Characteristics of the structure and function of male reproductive organs in the copulating sculpin Radulinopsis taranetzi were investigated based on histological observations. The male reproductive organs comprised three parts: a pair of testes, a seminal vesicle, and a penis. Germinal cells matured in cysts located in the small seminal lobules. Asynchronous spermatogenesis advanced rapidly from the posterior to the anterior region of the testes. After sperm matured in the posterior part of the testes, the seminiferous epithelium of the seminal lobules synthesized and secreted eosinophilic fluid that showed a positive periodic acid–Schiff (PAS) reaction into the seminal lobules. Spermatozoa excreted from the posterior part of the testes were stored together with the secretion in the seminal vesicle and showed no activity in the seminal fluid. Histological observations throughout the year suggest that the fluid is secreted and spermatozoa are stored in the seminal vesicles during February to July, which is presumably when mating occurs. The importance of testicular maturation and the secretion of eosinophilic fluid during this long reproductive period is also discussed.     

Effects of nutrition and methoprene treatment upon reproductive diapause in Caloptilia fraxinella (Lepidoptera: Gracillariidae)

Abstract Female Caloptilia fraxinella exhibit a prolonged reproductive diapause immediately post adult emergence in mid‐summer until the next spring when mating, egg development and oviposition on fresh Fraxinus spp. leaflets occur. Factors that effect the termination of reproductive diapause are investigated in this species. Caloptilia fraxinella diapausing adults held in overwintering conditions (2 °C, LD 0 : 24 h) for 24 weeks terminate diapause after placement for 2 weeks in simulated summer conditions (24 °C, LD 16 : 8 h) only if they are provided with 10% sugar water. Exogenous application of the Juvenile Hormone (JH) analogue methoprene to moths in both early‐ (summer) and mid‐ (autumn) reproductive diapause demonstrates that JH affects diapause termination but a carbohydrate nutrition source also mediates mating and vitellogenesis. Mating between moth pairs early in diapause occurs only after treatment with methoprene and provision with sugar water. However, there is no impact of mating on the propensity of females to produce vitellogenic oöcytes. Moths collected in the autumn in mid‐diapause respond in a dose‐dependent fashion to methoprene treatment and the response is greater than that of moths early in diapause collected in the summer. Treatment with methoprene and access to sugar water results in vitellogenic oöcytes in 18.75% of females from mid‐diapause moth pairs treated with 0.01 μg methoprene per insect and in all females from pairs treated at the two highest doses of methoprene (0.1 and 1 μg per insect). Mating occurs only between moths in mid‐diapause treated with the two highest doses of methoprene and these doses induce 91% and 100% mating, respectively. Both control and methoprene‐treated males in mid‐diapause held under summer conditions mate successfully and pass a spermatophore to their methoprene‐treated female partner. These data demonstrate that female C. fraxinella undergo a prolonged reproductive diapause in which termination is dependent on JH and further mediated by a carbohydrate nutrition source. The production of vitellogenic oöcytes is independent of mating. These data also provide evidence that response of moths in diapause to exogenous applications of methoprene differs throughout the diapause period and between male and female C. fraxinella.     

Expression of cysteine sulfinate decarboxylase (CSD) in male reproductive organs of mice

Cysteine sulfinate decarboxylase (CSD) is the rate-limiting biosynthetic enzyme of taurine, but it is still controversial whether the male reproductive organs have the function to synthesize taurine through CSD pathway. The present study was thus undertaken to detect CSD expression in male mouse reproductive organs by RT-PCR, Western blot and immunohistochemistry. The results show that CSD is expressed both at the mRNA and protein levels in the testis, epididymis and ductus deferens. The relative levels of both CSD mRNA and protein increase from the testis to the epididymis and to the ductus deferens. Immunohistochemical results demonstrate that the main cell types containing CSD are Leydig cells of testis, epithelial cells and some stromal cells throughout the efferent ducts, epididymis and ductus deferens. These results suggest that male genital organs have the function to produce taurine through the CSD pathway, although quantifying the relation of CSD expression to taurine synthesis and the exact functions of taurine in male genital organs still need to be elucidated in future studies.