Efferent Ducts Research Articles

Epididymal morphophysiology of adult collared peccaries (Pecari tajacu)

O entendimento funcional do epidídimo possibilita o aumento da fertilidade de uma espécie, pois torna possível a preservação de gametas para o uso em técnicas de reprodução assistida. Este trabalho estudou os aspectos histológicos das diferentes regiões do epidídimo de Pecari tajacu, bem como as patologias associadas encontradas nos espermatozoides presentes nesse órgão. Após procedimento cirúrgico de orquiectomia, para remoção dos testículos e epidídimos, obtidos de oito animais adultos nascidos em cativeiro, realizou-se a fixação e o processamento histológico das amostras. Constatou-se em todas as regiões do epidídimo (cabeça, corpo e cauda) a presença de epitélio pseudoestratificado colunar com estereocílios. Foram observadas semelhanças entre a altura média do epitélio do corpo e da cauda, a qual era diferente da encontrada na cabeça (1- proximal: região associada aos ductos eferentes; 2- distal: região associada ao corpo do epidídimo). Foi verificado que a cabeça proximal apresenta a média de diâmetro dos túbulos menor em relação às demais porções do epidídimo. Em relação às patologias dos espermatozoides, constataram-se treze tipos diferentes de alterações morfológicas. Os espermatozoides subdesenvolvidos e cabeças soltas foram as mais frequentes ao longo do epidídimo de catitus. Os dados deste trabalho são pioneiros, e servirão como base para pesquisas comparativas sobre a biologia reprodutiva dos artiodátilos, contribuindo para a sua conservação e produção.

Morpho-anatomy of the male reproductive tract and spermatogenesis of the South American Spasalus silvarum Kuwert (Coleoptera: Passalidae)

The morpho-anatomy of the reproductive tract and of the germ cells has been useful to establish phylogenetic relationships in many insects. However, these elements remain little known in the Coleoptera Passalidae. Here, the male reproductive tract, the spermatogenesis and the sperm structure are described in the South American Spasalus silvarum Kuwert, 1898 (Passalinae, Passalini), a subsocial species living under decaying bark. In this beetle, the two fusiform follicles of each testicle have a central intrafollicular efferent duct and are divided in septa. This organization of the cysts is different from those known in other insects. Each cyst contains up to 128 filiform spermatozoa with relatively small nuclei (29 µm in length) in relation to the length of the flagellum (880 µm). Among passalids, the general conformation of the male reproductive tract is similar to that described in other American species, but it differs from the African ones in terms of number and shape of testicular follicles. The data provided from the spermatozoa and from their maturation process contribute to the understanding of the reproductive cell biology of Passalidae.

Intersex and liver alterations induced by long-term sublethal exposure to 17α-ethinylestradiol in adult male Cnesterodon decemmaculatus (Pisces: Poeciliidae).

The aim of the present study was to assess the responses of the gonopodium morphology and the gonadal and liver histology of adult male Cnesterodon decemmaculatus to sublethal long-term exposure concentrations of 17α-ethinylestradiol (EE2). Two experiments were conducted exposing the fish to waterborne concentrations of EE2 ranging from 20 ng/L to 200 ng/L for 8 wk, 12 wk, and 16 wk. Intersex gonads were observed after 8 wk and 16 wk in fish exposed to 200 ng EE2/L and 100 ng EE2/L, respectively. Oocytes' development from testis germ cells and replacement of the efferent duct periodic acid-Schiff-positive secretion surrounding spermatozeugmata by parenchymal tissue and duct structure alterations were the major observed changes in the gonads. In contrast, no response was observed in the gonopodium morphology. Liver histology was also altered, showing increasing steatosis, single-cell necrosis to generalized necrosis, and disruption of acinar organization from 100 ng EE2/L to 200 ng EE2/L. In summary, the present results showed that although EE2 was not able to alter the morphology of a developed gonopodium, it was capable of inducing development of testicular oocytes in adult male C. decemmaculatus at environmentally relevant concentrations. Thus, externally normal but intersex C. decemmaculatus males would be expected in the wastewater-receiving streams that the species inhabits. According to the literature, the present study would be the first indicating estrogen-induced intersex in adult male poeciliid. Environ Toxicol Chem 2017;36:1738-1745. © 2016 SETAC.

Comprehensive Immunophenotypic Characterization of Adult and Fetal Testes, the Excretory Duct System, and Testicular and Epididymal Appendages.

The immunophenotype of a normal testis and the excretory duct system has not been studied comprehensively in fetal and adult patients without testicular disease or hormonal manipulation so far. In addition, testicular (TA) and epididymal (EA) appendages are frequent paratesticular structures without previously reported comprehensive immunophenotypic studies. Immunohistochemistry for multiple markers, including the androgen receptor (AR), the estrogen receptor (ER), the progesterone receptor (PR), the prostate-specific antigen, the prostate-specific membrane antigen, PAX8, WT1, calretinin, CK7, CK20, OCT4, SALL4, and CD117, was performed on full sections of testicular/paratesticular tissue from a large cohort of adult and fetal autopsy patients. In contrast to adult germ cells (GC), fetal GC strongly express OCT4 and CD117, although the expression of these proteins is lost in the early postnatal period; SALL4, in contrast, is expressed in both fetal and adult GC, with only weak and focal expression in adult patients. Fetal Sertoli cells (SC) express WT1 and calretinin strongly and diffusely, in contrast to adult SC. Both fetal and adult excretory duct systems express CK7 and PAX8 with frequent AR coexpression, and all 3 main segments of the excretory duct system (ductuli efferentes, epididymis, and vas deferens) have unique immunophenotypes. The rete testis also has a unique immunohistochemical expression pattern, which includes strong expression of CK7, PAX8, WT1, calretinin, and AR. Finally, of the adult autopsy patients examined, 80% had a TA, and 60% had an EA; these paratesticular structures occurred at stereotypical locations, demonstrated reproducible morphologic features, and had a unique immunophenotype relative to other studied structures, with strong CK7, PAX8, WT1, AR, ER, and PR coexpression. The testis and the paratestis may be involved by diverse neoplastic and non-neoplastic processes, and knowledge of the immunophenotypic expression spectrum of these tissues may aid in clinical diagnosis and advance our understanding of the pathogenesis of both oncologic and nononcologic disease processes.

Identification of NR0B1 as a novel androgen receptor co-repressor in mouse Sertoli cells.

Nuclear receptor subfamily 0 group B member1 (Nr0b1) is an atypical member of the nuclear receptor family that is predominantly expressed in mouse Sertoli cells(SCs). Mutations of NR0B1 in humans cause adrenal failure and hypogonadotropic hypogonadism. The targeted mutagenesis of Nr0b1 in mice has revealed a primary gonadal defect characterized by the overexpression of aromatase and cellular obstruction of the seminiferous tubules and efferent ductules, leading to germ cell death and infertility. The transgenic expression of Nr0b1 under the control of the Müllerian-inhibiting substance promoter(MIS-Nr0b1), which is selectively expressed in SCs, improves fertility. Testicular androgen receptor(AR) was also expressed in SCs. Many genes are directly regulated by androgen and its AR, which are involved in spermatogenesis and male infertility. As the association between NR0B1 and AR remains unclear in mouse SCs, we decided to further explore the relationship between them. In the present study, we have identified NR0B1 as a novel AR co-repressor in mouse SCs. Using RT‑qPCR and immunofluorescence, we determined that NR0B1 was mainly expressed in mouse SCs in an age-dependent manner from 2-8weeks of age postnatally. The inhibition of the effects of AR on AR target genes by NR0B1, in an androgen‑dependent manner, was further demonstrated by western blot analysis and RT-qPCR in TM4 cells, a mouse Sertoli cell line. Finally, invitro luciferase and co-immunoprecipitationassays validated that NR0B1, as an AR co-repressor, significantly inhibited the transcriptional activation of its target genes. These results suggest that novel inhibitory mechanisms underlie the effects of NR0B1 in modulating androgen-dependent gene transcription in mouse SCs.

Stem Cell Therapy for Male Infertility: A Review

Infertility affects 15% of couples in reproductive age world-wide and male factor is solely responsible in about 50% of the cases and contributory in 30-40% of cases. Spermatogonial stem cells (SSCs) are small self-renewing cells found in the basal compartment of seminiferous tubules where they form the foundation of spermatogenesis and are required for the continuous production of sperm. Transplantation of spermatogonial stem cells results in a donor derived sperm production and fertility in rodent and non-rodent species like Goat. Transplantation of cryopreserved spermatogonial stem cells could help oncology patients undergoing radiotherapy or chemotherapy by prior isolation of the SSCs and transplantation after treatment. Induced pluripotent stem cells also have the ability to differentiate into late stage germ cells. The efficacy and safety of SSCs transplantation showed that offspring produced did not show any morphological differences or alteration of genetic material but is most effective through assisted reproduction and better on young/immature Animals. The transfer of germ cells through micro-injection of seminiferous tubules and cannulation of efferent ducts is most effective on rodent testis while injection into the rete testis guided by ultrasound was reported to be the best technique in higher Animals (Bulls and Apes). Researches are still ongoing to get a safe and successful method of SSCs transplantation with no or less side effects on humans.Keywords: Cryopreserved, Pluripotent, self-renewing, Spermatogonia, and Spermatogenesis

PRENATAL DEVELOPMENT OF THE RETE TESTIS AND MEDIASTINUM TESTIS IN DROMEDARY CAMEL (CAMELUS DROMEDARIES)

The present investigation was carried out to demonstrate the sequential stages of the prenatal development of the rete testis and mediastinum testis by histological methods. Ninety eight male camel embryos with varying CVRL ranging from (2.2 cm -105 cm) were used in the current study. The indifferent gonads appeared in embryos of 2.2 cm CVRL in the form of two small bilateral cellular masses medial to the mesonephrii. The parenchyma of the indifferent gonads didn’t show any pattern of cord like arrangement. ). In embryos of 4 cm CVRL embryos the primordia of the tunica albuginea was detected in the form of thin layer of mesenchymal cells surrounding the developing gonad. In embryos of 6.5 cm CVRL the primordia of the presumptive mediastinum testis was evident nearby the mesorchium in the form of delicate fibrous and vascular structure. In embryos of 23-25 cm CVRL the central portion of the testis was clearly devoid of the testicular cords, but equipped with the vasal tubules of the rete testis and the mediastinum which was extended in the form of an axial cord within the centrum of the testis. In embryos of 30-32 cm CVRL the straight radicals of rete testis revealed anastomosis with the neighboring testicular cords. In embryos of 69 cm CVRL the mediastinum testis revealed more fibrous component of interlacing and corrugated collagenic fibers that coursed in the form of strand like bundles in-between the branches of the rete testis. Moreover, the mediastinum was colonized by cellular aggregations of mitotically active interstitial endocrine cells. The connection between the tubules of the rete testis and the efferent ductules was observed in embryos of 88 cm CVRL near the head pole of the testis. The tubules of the rete testis were found to be lined with single cell layer of cuboidal epithelium throughout the whole period studied ,

The human epididymis: its function in sperm maturation.

Spermatozoa acquire their fertilizing ability and forward motility properties during epididymal transit. Our knowledge of gamete physiology is based on studies conducted in laboratory and domestic species; our knowledge of these processes in humans is limited. Medical indications for assisted reproductive technologies (ART) have progressed to include male infertility. Surgical procedures allow collection of spermatozoa from all along the human excurrent ducts, and the former have been used with some success in reproductive medicine. This has raised questions over the role of the epididymis in human sperm physiology. To reanalyze what we now know about epididymal physiology in humans and to assess the relevance of laboratory animal models for understanding human physiology and the pathophysiology of the epididymis. A systematic bibliographic search of PubMed for articles published in English before May 2015 was carried out using the search terms 'epididymis' and 'sperm maturation'. Literature on the consequences of vasectomy on the epididymis was also searched. Whereas the proximal epididymis is almost exclusively occupied by efferent ducts, the sperm reservoir capacity is poorly developed in humans. At the molecular level, the human transcriptome and proteome show some segment specificity; conflicting results persist with regard to secretome variation along the tubule. The number of genes regulated along the excurrent ducts in men is lower when compared to rodent species, but remains significant. It is challenging to reconcile biochemical and physiological studies with clinical data obtained from men undergoing reanastomosis of the vas deferens at different points along the excurrent duct. We propose that vasectomy/vasovasostomy is a model to understand the consequences of obstruction on epididymis function in humans. Despite the scarcity of biological material available, the interspecies variability of the male reproductive tract urges us to use modern molecular and cellular biology tools to better understand human epididymis physiology in order to apply ART in a more responsible manner.

Seasonal and Ageing-Depending Changes of Aquaporins 1 and 9 Expression in the Genital Tract of Buffalo Bulls (Bubalus bubalis).

The presence of Aquaporins 1 (AQP1) and 9 (AQP9), integral membrane water channels that facilitate rapid passive movement of water and solutes, was immunohistochemically detected in the excurrent ducts collected from sexually mature buffalo bulls of proven fertility during the mating (late autumn-winter) and non-mating (late spring to the beginning of autumn) seasons. Furthermore, the research was performed also on the epididymal cauda of a senile buffalo bull with inactive testis. Aquaporins 1 and 9 were immunolocalized at distinct levels. In the efferent ducts, AQP1 immunoreactivity was strongly evidenced at the apical surface of the non-ciliated cells and weakly along the basal membrane of the epithelial cells. The latter reactivity disappeared during the non-mating season. No AQP1 immunoreactivity was detected in the epithelium of epididymis and vas deferens, whereas AQP1 was expressed in the smooth muscle layer of the vas deferens. Aquaporin 1 was present in the blood vessels and in small nerve bundles all along the genital tract. The supranuclear zone of the epididymal principal cells was AQP9 immunoreactive, limited to the corpus and cauda regions, and vas deferens. The samples collected in the two reproductive seasons showed a weaker AQP9 immunoreactivity during the non-mating season. A typical AQP9 immunoreactivity was noticed in the old buffalo examined. The tested AQP molecules showed a different expression pattern in comparison with laboratory mammals, primates, equine, dog and cat. In addition, seasonal differences were noticed which are possibly useful in regard to the comprehension of the morphophysiology of reproduction in the bubaline species, which are still a matter of debate.

Histomorphometric evaluation of treatment of rat azoosper-mic seminiferous tubules by allotransplantation of bone marrow-derived mesenchymal stem cells.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) potentials make them appropriate for cell therapy including ability of differentiation and release of anti-inflammatory cytokines and growth factors secreta. For treatment of azoospermia to induce proliferation and differentiation of germ cells, MSCs transplantation has been introduced. The aim of the present experimental case-control study was to histomorphometric evaluation of the germinal cells in seminiferous tubules of azoospermic rats before and after BM-MSCs allotransplantation. In the present study, BM-MSCs were isolated from six male rats and confirmed. Their testes also served as intact negative controls. The recipient rats (n=6) were received two doses of 10 mg/kg of busulfan with 21 days interval to induce azoospermia. After cessation of spermatogenesis, the rats were allotransplanted with the BM-MSCs into efferent duct of right testes. Thirty-five days later, the right cell-treated testes were compared to left azoospermic ones. Histomorphometric analyses showed that the seminiferous tubules treated with BM-MSCs had normal morphology in comparison with azoospermic testes, which were without germinal layer. In most BM-MSCs-treated seminiferous tubules, spermatogenesis was observed. The allotransplanted BM-MSCs could induce spermatogenesis in seminiferous tubules of azoospermic rats.

Membrane-Localized Estrogen Receptor 1 Is Required for Normal Male Reproductive Development and Function in Mice.

Estrogen receptor 1 (ESR1) mediates major reproductive functions of 17β-estradiol (E2). Male Esr1 knockout (Esr1KO) mice are infertile due to efferent ductule and epididymal abnormalities. The majority of ESR1 is nuclear/cytoplasmic; however, a small fraction is palmitoylated at cysteine 451 in mice and localized to cell membranes, in which it mediates rapid E2 actions. This study used an Esr1 knock-in mouse containing an altered palmitoylation site (C451A) in ESR1 that prevented cell membrane localization, although nuclear ESR1 was expressed. These nuclear-only estrogen receptor 1 (NOER) mice were used to determine the roles of membrane ESR1 in males. Epididymal sperm motility was reduced 85% in 8-month-old NOER mice compared with wild-type controls. The NOER mice had decreased epididymal sperm viability and greater than 95% of sperm had abnormalities, including coiled midpieces and tails, absent heads, and folded tails; this was comparable to 4-month Esr1KO males. At 8 months, daily sperm production in NOER males was reduced 62% compared with controls. The NOER mice had histological changes in the rete testes, efferent ductules, and seminiferous tubules that were comparable with those previously observed in Esr1KO males. Serum T was increased in NOER males, but FSH, LH, and E2 were unchanged. Critically, NOER males were initially subfertile, becoming infertile with advancing age. These findings identify a previously unknown role for membrane ESR1 in the development of normal sperm and providing an adequate environment for spermatogenesis.

Effect of long-term treatment with Kanefron® H on the risk factors of the urinary system, associated with idiopathic calcium urolithiasis

Urolithiasis - nephrolithiasis, causes the formation of calculi in the kidneys and efferent urinary ducts due to the substances that form calculi. Idiopathic calcium urolithiasis - the most common form of the disease. Modern herbal medicine can play an important role in primary and secondary prevention of stone formation. Combination Kanefron® herbal drug H (drops: Centaurii herba, 1,8 g / 100 mL; Levistici radix, 1,8 g / 100 mL; Rosmarini folium, 1,8 g / 100 mL; Bionorica SE, Neumarkt / Germany) approved as a drug of vegetable origin. The aim of this study was to investigate risk factors for renal stone formation and activity in patients with idiopathic calcium urolithiasis during long herbal medicine. Thirty-two patients with idiopathic calcium urolithiasis received a daily dose of the drug plant 50 drops three times a day for two months. In another study, 18 patients with similar inclusion criteria were treated for 4 months. In both treatment groups receiving combination herbal preparation contributed to the increase in diuresis and increasing the pH of urine. Within 1 month after treatment, the stone formation (UFL) have improved, which is considered a lower risk. Compared with the 2 = month follow-up in patients who received 4 = monthly treatment, noted a significant improvement in the observed indicators. Adverse events were observed.

Haploinsufficiency of SF-1 Causes Female to Male Sex Reversal in Nile Tilapia, Oreochromis niloticus.

Steroidogenic factor-1 (Sf-1) (officially designated nuclear receptor subfamily 5 group A member 1 [NR5A1]) is a master regulator of steroidogenesis and reproduction in mammals. However, its function remains unclear in nonmammalian vertebrates. In the present study, we used immunohistochemistry to detect expression of Sf-1 in the steroidogenic cells, the interstitial, granulosa, and theca cells of the ovary, and the Leydig cells of the testis, in Nile tilapia. Clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (Cas9) cleavage of sf-1 resulted in a high mutation rate in the F0 generation and a phenotype of gonadal dysgenesis and reduced steroidogenic cells in XX and XY fish. Sf-1 deficiency also resulted in decreased cytochrome P450, family 19, subfamily A, polypeptide 1a, forkhead box L2 expression, and serum estradiol-17β in XX fish. In XY fish, Sf-1 deficiency increased cytochrome P450, family 19, subfamily A, polypeptide 1a and forkhead box L2 expression but decreased cytochrome P450, family 11, subfamily B, polypeptide 2 expression and serum 11-ketotestosterone levels. 17α-methyltestosterone treatment successfully rescued the gonadal phenotype of Sf-1-deficient XY fish, as demonstrated by normal spermatogenesis and production of F1 mutants. In contrast, estradiol-17β treatment only partially rescued the gonadal phenotype of Sf-1-deficient XX fish, as demonstrated by the appearance of phase II oocytes. Furthermore, both sf-1(+/-) F1 XX and XY mutants developed as fertile males, although spermatogenesis was delayed and efferent duct formation was disordered. Our data suggest that Sf-1 is a major regulator of steroidogenesis and reproduction in fish, as it is in mammals. Sf-1 deficiency resulted in gonadal dysgenesis and feminization of XY gonads. However, unlike in mammals, Sf-1 deficiency also resulted in female to male sex reversal in 8.1% of F0 and 92.1% of sf-1(+/-) F1 in XX fish.

Salivary gland morphology, tissue tropism and the progression of tospovirus infection in Frankliniella occidentalis

Tomato spotted wilt virus (TSWV) is transmitted by thrips in a propagative manner; however, progression of virus infection in the insect is not fully understood. The goal of this work was to study the morphology and infection of thrips salivary glands. The primary salivary glands (PSG) are complex, with three distinct regions that may have unique functions. Analysis of TSWV progression in thrips revealed the presence of viral proteins in the foregut, midgut, ligaments, tubular salivary glands (TSG), and efferent duct and filament structures connecting the TSG and PSG of first and second instar larvae. The primary site of virus infection shifted from the midgut and TSG in the larvae to the PSG in adults, suggesting that tissue tropism changes with insect development. TSG infection was detected in advance of PSG infection. These findings support the hypothesis that the TSG are involved in trafficking of TSWV to the PSG.

Structural changes in female-to-male sex-reversing gonads of Rana RUGOSA.

The phenotypic sex of many species of amphibians is subject to reversal by steroid hormones. The mechanism of this process, however, still remains largely unknown. As a step toward understanding the histological changes during sex reversal in amphibians, we analyzed two- and three-dimensional (2D and 3D) structures of sex-reversing gonads in Rana rugosa frogs. 2D views revealed that many oocytes in the wild-type ovary disappeared during female-to-male sex-reversal concomitant with the emergence of Vasa-positive small germ cells. Some of the germ cells were labeled with BrdU. BrdU-positive germ cells were few in the testosterone (T) treated ovaries at days 8 and 16, which resembled wild-type ovaries. Basement membranes became disrupted by day 24 in T-treated ovaries. However, the membranes were later reconfigured into testis-like gonadal structures 40 days after T treatment. 3D imaging of the sex-reversing gonad using serial immunostained sections showed that germ cells were organized in linear fashion extending out from where the sex-reversing gonad attached to the mesorchium 24 days after T treatment. Germ cells were increased in number by 40 days and were localized to the cortex of the gonads. In a T-untreated testis at day 24, many germ cells were distributed throughout the cortex except in the central space, while the efferent duct ran between two sheets of the mesorchium. These results, taken together, suggest that the mesorchium plays an important role in the organization of testicular structure. This is the first report showing germ cell ontogeny and organization in the female-to-male sex-reversing gonad in a vertebrate species.

Testicular Busulfan Injection in Mice to Prepare Recipients for Spermatogonial Stem Cell Transplantation Is Safe and Non-Toxic.

Current methods of administering busulfan to remove the endogenous germ cells cause hematopoietic toxicity, require special instruments and a narrow transplantation time. We use a direct testicular injection of busulfan method for preparing recipients for SSC transplantation. Male ICR mice (recipients) were divided into four groups, and two experimental groups were treated with a bilateral testicular injection of 4 or 6 mg/kg/side busulfan (n = 60 per concentration group). Mice received an intraperitoneal injection (i.p.) of 40 mg/kg busulfan (n = 60, positive control) and bilateral testicular injections of 50% DMSO (n = 60, negative control). Donor SSCs from RFP-transgenic C57BL/6J mice were introduced into the seminiferous tubules of each recipient testis via efferent duct injection on day 16–17 after busulfan treatment. Recipient mice mated with mature female ICR mice and the number of progeny was recorded. The index detected at day 14, 21, 28, 35 and 70 after busulfan treatment. Blood analysis shows that the toxicity of busulfan treated groups was much lower than i.p. injection groups. Fertility was restored in mice treated with busulfan and donor-derived offspring were obtained after SSC transplantation. Our study indicated that intratesticular injection busulfan for the preparation of recipients in mice is safe and feasible.

Region-specific localization of NOS isoforms and NADPH-diaphorase activity in the intratesticular and excurrent duct systems of adult domestic cats (Felis catus).

Nitric oxide (NO) is produced by nitric oxide synthases (NOSs) and plays an important role in all levels of reproduction from the brain to the reproductive organs. Recently, it has been discovered that all germ cells and Leydig cells in the cat testis exhibit stage-dependent nuclear and cytoplasmic endothelial (eNOS) and inducible (iNOS)-NOS immunoreactivity and cytoplasmic nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) reactivity. As a continuation of this finding, in this study, cellular localization of NADPH-d and immunolocalization and expression of all three NOS isoforms were investigated in the intratesticular (tubuli recti and rete testis), and excurrent ducts (efferent ductules, epididymal duct and vas deferens) of adult cats using histochemistry, immunohistochemistry and western blotting. NADPH-d activity was found in the midpiece of the spermatozoa tail and epithelial cells of all of ducts, except for nonciliated cells of the efferent ductules. Even though the immunoblotting results revealed similar levels of nNOS, eNOS and iNOS in the caput, corpus and cauda segments of epididymis and the vas deferens, immunostainings showed cell-specific localization in the efferent ductules and region- and cell-specific localization in the epididymal duct. All of three NOS isoforms were immunolocalized to the nuclear membrane and cytoplasm of the epithelial cells in all ducts, but were found in the tail and the cytoplasmic droplets of spermatozoa. These data suggest that NO/NOS activity might be of importance not only for the functions of the intratesticular and excurrent ducts but also for sperm maturation.

The Male Urogenital System of the Siberian Salamander Salamandrella keyserlingii (Caudata: Hynobiidae) with Special Reference to the Microstructure of the Testes and Sperm Transport Complex

The gross anatomy of the urogenital system and histology of the testes and sperm transport system in adult male Salamandrella keyserlingii from southeast of Western Siberia were examined. The general structure of the male urogenital system in S. keyserlingii was similar to that of other hynobiids. Testes were monolobed and bean shaped. No spermatogenic wave through the longitudinal axis of the testes was observed. However, a «lobular wave» with zonal distribution of cysts at different sperm maturation stages was observed along the proximo-distal axis of each seminiferous lobule. From the testes to the cloaca, the sperm transport system included the vasa efferentia, longitudinal collecting ducts of the vasa efferentia, nephrons of the genital kidneys, collecting ducts, and Wolffian ducts.

Effect of Coumestrol on Efferent Ductules in Dogs

The present study was conducted on twelve adult stray dogs of age 2–3 years, weighing 14–18 kg and randomly divided into three groups. Animals in group I (n = 5) and group II (n = 5) were orally given 300 and 500 microgram coumestrol dissolved in di-methyl sulfoxide in the commercial dog treats on days 0, 7, 14, 21 and 28, respectively. Animals in control group III (n = 2) were given DMSO alone as per above schedule. Castration was done one week after the completion of the treatment. Tissues for histology and electron microscopy were fixed in Bouin's fixative and Karvonsky's fixatives, respectively. Epithelial lining of the efferent ductules was composed of cuboidal or columnar epithelium having ciliated and non-ciliated cells. Cellular architecture of the efferent ductules was normal. Scanning electron microscopy revealed normal efferent ductules with presence of cilia on luminal surface. In conclusion, oral feeding of 300 microgram and 500 microgram of coumestrol has no adverse effects on the functioning of efferent ductules in dogs and at this dose rate, this compound cannot be used for population control of stray dogs.

Morphological and ultrastructural study of the efferent ductules in the Chinese soft-shelled turtlePelodiscus sinensis

Comparative study of the turtle excurrent duct system increases our understanding the evolution of sperm motility and fertility maintenance in higher vertebrates. Therefore, in this study we observed the histology and ultrastructure organization of efferent ductules in the Pelodiscus sinensis using light and transmission electron microscopy. The efferent ductules are extra- testicular and 22-28 in number originate from rete testis. The epithelium is entirely composed of two types of cells, the predominant non-ciliated and ciliated cells. The ciliated cells have long cilia that protrude into the lumen to form a meshwork. These cells associated with clusters of mitochondria in the supranuclear cytoplasm and possess coated vesicles, vacuole, intracellular spaces, and junction complexes. Ciliated cells in the proximal portion of the ductules contain an endocytic apparatus with coated pits and tubules in the apical cytoplasm. Interdigitations and lipid droplets are predominantly present around the nuclei of these cells. The non-ciliated cells have clusters of mitochondria present in both the supranuclear and perinuclear cytoplasm whereas, the nuclei of these cells are lightly stained. Moreover, the contour of the epithelium towards lumen is irregular as it has a deep indentation. The apical cytoplasm goes deep into the lumen to form cytoplasmic processes. This is the first study to describe the detailed features of efferent ductules in Pelodiscus sinensis with, special focus on the morphology of ciliated cells, as these cells are involved in the mixing of luminal fluid and transport of spermatozoa towards the distal region.