Interstitial Leydig Cells Research Articles

Regulated and Functional Expression of the Corepressor MTA3 in Rodent Testis.

Metastasis-associated protein (MTA)3 is a subunit of the Mi-2/nucleosome remodeling and deacetylase protein complex, with relevant roles in the regulation of cancerous epithelial to mesenchymal transition in an estrogen-dependent manner, recently involved in the modulation of different physiological processes. Although these findings connect MTA3 expression with hormonal signaling in various systems, little is known about whether this relationship is conserved in testis where hormonal action is intensive. We, therefore, document here for the first time the expression of Mta3/MTA3 in mammalian testes, where MTA3 protein was identified mainly in interstitial Leydig cells. Testicular levels of Mta3/MTA3 were overtly modulated by pituitary gonadotropins, as well as metabolic signals, such as dexamethasone, T4, and rosiglitazone. In addition, ablation of endogenous Mta3 by short hairpin RNA significantly inhibited human choriogonadotropin/dibutyryl-cAMP (db-cAMP)-stimulated progesterone secretion in MA-10 Leydig cells, whereas overexpression of exogenous MTA3 effectively reversed Mta3 deficiency damaged progesterone production. Moreover, attenuated Mta3 expression positively correlated to the deregulated level of serum testosterone in murine type 2 diabetes mellitus. From a functional standpoint, MTA3 deficiency was involved in insulin-mediated inhibition of testicular steroidogenesis. Our data are the first to disclose the presence and functional role of MTA3 in the testis, where its expression is regulated by developmental, metabolic, and hormonal cues and is closely associated with steroidogenic dysfunction. The current study expands the reproductive dimension of MTA3, which may operate directly at the testicular level to modulate steroidogenic function.

ПАТОМОРФОЛОГІЧНІ ОСОБЛИВОСТІ ЛЕЙДИГОМИ СОБАК

The research for the study of morphological, clinical and macroscopic characteristics of the different histological types of canine testicular tumors. Material was obtained during a routine surgery for the removal of testicular tumors in dogs in clinics of veterinary medicine in Kiev. Tumors were divided into sex-cord stromal tumors and germ cell tumors. Among the sex-cord stromal tumors recorded leydig (interstitial) and sertoli cell tumors. Macroscopical and histologic characteristics of canine interstitial (Leydig) cell tumor are described. It is 12.5% of all testicular tumors. The average age of dogs is 9 years. Largest share is breeds dog such as foxterriers and toyterriers. Interstitial (Leydig) cell tumor has a slower growth compared to other tumors the testicle, no metastases. Half the animals had no clinical signs. Mostinterstitial (Leydig) cell tumor go together with the pathology of adnexal structures. This is because Leydig cell tumor lead to increased levels of androgens. This fact may cause pathology of adnexal structures. Neoplasms are recorded both on the right and the left testis. Leydig cell tumor was not related to the cryptorchid testicles. Most tumors regular oval, brown or gray color, different texture and size with no signs of necrosis or ulceration. The average volume of tumors ranged from 2 to 20 cm2. Microscopically, the tumor is composed of cells that resemble Leydig cells and that are arranged into islands or tubular structures. Find polygonal eosinophilic cells with granular or vacuolated, which contains lipids. Nuclei were round mesh of chromatin and distinct nucleolus. Sometimes noted the large cells with 1, 2 or more nucleoli. In some cases, found elongated fusiform cells with small nuclei and granular eosinophilic cytoplasm. Tumor stroma characterized by calcification. Note mitotically active cells. Cysts lined by tumor cells are present in some tumors. Distinction between benign and malignant forms on purely histological criteria, in the absence of metastasis, often is difficult. Studies on the features of histological structure of testicular tumors will optimize the histological diagnosis of neoplasms.

Urocortinergic system in the testes of normal and cryptorchid dogs

Cryptorchidism is the most common disorder of the sexual development in dogs, occurring in 13% of the males. Unilateral cryptorchidism is more frequent than bilateral and the right testis seems to be more frequently affected. Urocortin (UCN) is a corticotrophin-releasing hormone (CRH)-related peptide which was observed to affect several functions in male genital organs. The aim of the present study was to investigate the expression of UCN, and its receptors CRHR1 and CRHR2 by immunohistochemistry, Western blot and real-time RT-PCR in the normal and cryptic testis of the dog. The results showed that UCN, CRHR2 and CRHR1 were expressed in normal and cryptic testes. UCN-immunoreactivity (IR) was distributed in germ cells of the normal and cryptic testis. In the normal testis, CRHR2-IR was found in germ and interstitial Leydig cells. In the cryptic testis CRHR2-IR was distributed in gonocytes and interstitial Leydig cells. CRHR1-IR was distributed in the vessel smooth musculature and peritubular myoid cells. UCN and CRHR2 mRNA expression levels were lower in the cryptic than in normal testes. These results suggest that UCN and its receptors might play a role in regulating the spermatogenesis and hormonal activity of interstitial Leydig cells of the dog testis.

Age-Related Changes in the Histological Structure of Rabbit Testes and the Prophylactic Role of Vitamin E.

The present investigation is planned to compare between the histological structure of rabbit testes of different ages and the prophylactic role of vitamin E on senescent animals. Male NewZeland rabbits were divided into four groups according to age. Group I: represented the young rabbits (one month age, weighing 1 ± 0.4 kg), group II: adult rabbits (6 months age, weighing 4 ± 0.5 kg), group III: aged animals (24 month age, weighing 7.5 ± 0.5 kg and group (IV) aged rabbits treated daily with the therapeutic dose of vitamin E orally at a dose of 10 mg/kg b.w / day for 60 days. The testes of young rabbits revealed a small form of seminiferous tubules with regular outline with spermatogonia and Sertoli cells, and the lumen appeared empty from spermatozoa. The interstitium is made up of loose connective tissue with large polyhedral interstitial Leydig cells. In adult studied animals, testes demonstrated with all spermatogenic cells and the lumen filled with spermatozoa. In aged rabbits, the histological studies of testis demonstrated atrophied irregular tubules, disarrangement of vacuolated spermatogenic cells, and sloughing of germ cells into the tubular lumen with reduction and degeneration of spermatozoa. The interstitium between the tubules was widen and filled with degenerated and reduced Leydig cells. Administration of vitamin E to aged animals group revealed an obvious improvement of spermatogenic cells, and the spermatozoa appeared in the lumen of many seminiferous tubules.

NNT mutations: a cause of primary adrenal insufficiency, oxidative stress and extra-adrenal defects.

Nicotinamide nucleotide transhydrogenase (NNT), one of the several genes recently discovered in familial glucocorticoid deficiencies (FGD), is involved in reactive oxygen species detoxification, suggesting that extra-adrenal manifestations may occur, due to the sensitivity to oxidative stress of other organs rich in mitochondria. Here, we sought to identify NNT mutations in a large cohort of patients with primary congenital adrenal insufficiency without molecular etiology and evaluate the degree of adrenal insufficiency and onset of extra-adrenal damages. Sanger or massive parallel sequencing of NNT and patient monitoring. Homozygous or compound heterozygous NNT mutations occurred frequently (26%, 13 unrelated families, 18 patients) in our cohort. Seven new mutations were identified: p.Met337Val, p.Ala863Glu, c.3G>A (p.Met1?), p.Arg129*, p.Arg379*, p.Val665Profs*29 and p.Ala704Serfs*19. The most frequent mutation, p.Arg129*, was found recurrently in patients from Algeria. Most patients were diagnosed belatedly (8-18 months) after presenting severe hypoglycemia; others experiencing stress conditions were diagnosed earlier. Five patients also had mineralocorticoid deficiency at onset. One patient had congenital hypothyroidism and two cryptorchidism. In follow-up, we noticed gonadotropic and genitalia impairments (precocious puberty, testicular inclusions, interstitial Leydig cell adenoma, azoospermia), hypothyroidism and hypertrophic cardiomyopathy. Intrafamilial phenotype heterogeneity was also observed. NNT should be sequenced, not only in FGD, but also in all primary adrenal insufficiencies for which the most frequent etiologies have been ruled out. As NNT is involved in oxidative stress, careful follow-up is needed to evaluate mineralocorticoid biosynthesis extent, and gonadal, heart and thyroid function.

Studies of gonadal sex differentiation

ABSTRACTGonadal differentiation has a determinative influence on sex development in human embryos. Disorders of sexual development (DSD) have been associated with persistent embryonal differentiation stages. Between 1998 and 2015, 139 female patients with various (DSD) underwent operations at the Scientific Center of Obstetrics, Gynaecology and Perynatology in Moscow, Russia. Clinical investigations included karyotyping, ultrasound imaging, hormonal measurement and investigations of gonadal morphology. The male characteristics in the embryo are imposed by testicular hormones. When these are absent or inactive, the fetus may be arrested at between developmental stages, or stay on indifferent stage and become phenotypically female. A systematic analysis of gonadal morphology in DSD patients and a literature review revealed some controversies and led us to formulate a new hypothesis about sex differentiation. Proliferation of the mesonephric system (tubules and corpuscles) in the gonads stimulates the masculinization of gonads to testis. Sustentacular Sertoli cells of the testes are derived from mesonephric excretory tubules, while interstitial Leydig cells are derived from the original mesenchyme of the mesonephros. According of the new hypothesis, the original mesonephric cells (tubules and corpuscles) potentially persist in the ovarian parenchyma. In female gonads, some mesonephric excretory tubules regress and lose the tubular structure, but form ovarian theca interna and externa, becoming analogous to the sustentacular Sertoli cells in the testis. The ovarian interstitial Leydig cells are derived from intertubal mesenchyme of the mesonephros, similar to what occurs in male gonads (testis). Surprisingly, the leading determinative factor in sexual differentiation of the gonads is the mesonephros, represented by the embryonic urinary system.

Oxytocin, Functions, Uses and Abuses: A Brief Review

Oxytocin a peptide hormone has the distinction of being the first among the peptide hormones to be sequenced and biochemically synthesized. Oxytocin is secreted from neural as well as non-neural sources inside the body. Among the neural sources supra-optic and paraventricular nuclei of the brain are major sources of secretion, the arcuate nucleus being a minor source. There are many non-neural sources from which synthesis of Oxytocin has been reported. The important ones include interstitial cells of Leydig in testis, epididymis, prostate glands, corpus luteum inside ovary, uterus, oviduct, placenta and adrenal glands. Oxytocin directly acts on the smooth muscle cells and causes muscle contraction. Indirectly it stimulates synthesis of PGF2α inside the female genital tract. As such the Oxytocin is involved in a wide spectrum of activities from milk letdown, parturition, penile erection and ejaculation to social, sexual and maternal behavior. Because of the spectrum of its functions it has since been used therapeutically for conditions like facilitating parturition in uterine inertia, prevent bleeding after parturition and abortion, management of incomplete abortion, milk letdown, retention of placenta etc., although the success has not been predictable. However, Oxytocin is also being misused under the false impression that its use in dairy animals increases the milk yield. Oxytocin abuse is also rampant in fruits and vegetables. This article is an attempt to explore the body of knowledge with respect to functions, uses and most importantly the abuses of this wonderful hormone.

70 GONADAL DEVELOPMENT IN GUINEA PIG MALES (CAVIA PORCELLUS)

Sexual differentiation in mammals is an event that presents many variations between species. Because it is related to hormonal function, any imbalance in the androgens and estrogens production can lead to malformations. Because sexual differentiation occurs in different ways among various animals, the recognition of their peculiarities becomes important in order to correct reproductive handling in different species. Considering that the guinea pig is commonly used as an experimental model in the reproductive area, the goal of this work was to perform a morphological description of gonad differentiation of the male guinea pig during embryonic development. In total, 11 conceptuses with ages 25 (n = 3), 30 (n = 2), 40 (n = 2), 50 (n = 2), and 65 (n = 2) days were used for light microscopy processing. The embryos at 25 days were processed completely. For the others, the gonads were dissected. The samples were dehydrated in alcohol, embedded in paraffin, and 5-µm sections were stained with hematoxylin-eosin. In the guinea pig gonad at 25 days gestation, there was a presence of gonadal cords, formed by condensation of somatic cells, which is characteristic of an undifferentiated gonad. In addition, we observed the presence of mesonephric and paramesonephric ducts in different embryos, indicating that other genital system organs were not formed. For the 30 days of development of guinea pigs, we observed that gonadal cords were differentiated in testicular cords by invasion of mesenchymal and endothelial cells, and also composed of Sertoli cells and primordial germ cells. These cords were among a large amount of testicular mesenchyme at the 40-day group. With 50- and 65-day development samples, the gonad was completely differentiated into testicle, with the presence of spermatogonia and Sertoli cells in the seminiferous tubules, and a large amount of interstitial Leydig cells around the tubules. We conclude that gonadal differentiation in guinea pig males occurs around the middle of pregnancy, between 25 and 30 days and that, before the end of the pregnancy, at 50 days, the testicle presents morphology similar to that found in the postnatal period.

Quantitative and cytological studies of interstitial (Leydig) cells in the scrotal and retained testes of unilateral cryptorchid West African Dwarf goats

Cryptorchidism is a common condition in West African Dwarf goats in Eastern Nigeria. The cytology and morphometry of interstitial cells were investigated using light and electron microscopy. Twenty adult male goats were used for the study. Ten of the goats were natural unilateral cryptorchids, while the remaining ten had fully descended testes and served as control. Significant hyperplasia of the interstitial cells were observed in scrotal and retained testes of the unilateral cryptorchid bucks compared to the control (normal) bucks. The number of interstitial cells per cross section of the testes was also significantly higher (P<0.05) in the contralateral scrotal testes of the unilateral cryptorchids (757.88±29.92) than in the testes of the normal bucks (677.30±25.92). In the contralateral scrotal testes of the unilateral cryptorchids, the interstitial cells had abundant cytoplasmic organelles, particularly mitochondria, Golgi apparatus and smooth endoplasmic reticulum, while the retained testes showed evidence of degeneration characterized by irregular nuclear profile, accumulation of nuclear heterochromatin, abundant lipid droplets, cytoplasmic vacuolations and focal intercellular dilations of interstitial cells. These results suggest that the intra-abdominal testes were significantly degenerated and the impairment was probably from exposure of the testes to higher abdominal temperature. However, the interstitial cells of the contralateral scrotal testes of the unilateral cryptorchid bucks showed normal histology and compensatory hyperplasia.

Slc15a1 is involved in the transport of synthetic F5-peptide into the seminiferous epithelium in adult rat testes.

Spermiation and BTB restructuring, two critical cellular events that occur across seminiferous epithelium in mammalian testis during spermatogenesis, are tightly coordinated by biologically active peptides released from laminin chains. Our earlier study reported that F5-peptide, synthesized based on a stretch of 50 amino acids within laminin-γ3 domain IV, could reversibly induce the impairment of spermatogenesis, disruption of BTB integrity, and germ cell loss, and thus is a promising male contraceptive. However, how F5-peptide when administered intratesticularly enters seminiferous tubules and exerts effects beyond BTB is currently unknown. Here we demonstrated that Slc15a1, a peptide transporter also known as Pept1, was predominantly present in peritubular myoid cells, interstitial Leydig cells, vascular endothelial cells and germ cells, while absent in Sertoli cells or BTB site. The steady-state protein level of Slc15a1 in adult rat testis was not affected by F5-peptide treatment. Knockdown of Slc15a1 by in vivo RNAi in rat testis was shown to prevent F5-peptide induced disruptive effects on spermatogenesis. This study suggests that Slc15a1 is involved in the transport of synthetic F5-peptide into seminiferous epithelium, and thus Slc15a1 is a novel target in testis that could be genetically modified to improve the bioavailability of F5-peptide as a prospective male contraceptive.

PPAR γ agonist pioglitazone partially reverses hyperglycemia but improves semen quality and testicular histomorphometrics in alloxan-induced diabetic rats

Abstract Introduction: Chronic hyperglycaemia is associated with subfertility and infertility in male diabetic subjects, possible as a result of perturbed testicular antioxidant defence system. Recent evidence however shows that some testicular cells produce insulin and this raises the possibility of a role for this hormone in spermatogenesis. Moreover, isoforms of the nuclear peroxisome proliferator activated receptor (PPAR) have been detected in testicular tissue, suggesting a role for insulin sensitization in testicular function. Materials and Methods: In this study, we report the ameliorative effects of the PPAR γ agonist pioglitazone on testicular morphologic aberrations and impaired semen profile induced by diabetes in rats treated with intraperitoneal alloxan (150 mg/kg). Diabetic rats were randomly assigned to receive oral pioglitazone at 10 or 30 mg/kg once daily for 6 weeks. Weekly blood glucose measurement using the glucometer showed weak effect of pioglitazone on hyperglycaemia in our model. Results: Nonetheless, findings from caudal epididymal sperm analysis showed significantly high sperm density in the pioglitazone-treated rats at the two doses tested (P &lt; 0.05 compared with diabetic controls). Moreover, pioglitazone at 10 mg/kg improved sperm motility and lowered the percentages of deformed and dead sperm cells in the treated diabetic rats. Haematoxylin and eosin (H&amp;E) sections of the testes showed signiicantly reduced seminiferous tubule diameter and lumen size, as well as diminished height of the germinal epithelium in the untreated diabetic rats, but not in rats on pioglitazone intervention. Furthermore, Gordon and Sweet section of the testes showed thickened interstitial compartment only in the untreated diabetic group. Meanwhile, interstitial cells of Leydig appeared undisturbed in testicular sections of all the groups, while intratesticular testosterone levels measured by the enzyme immunoassay technique were not significantly different between the treated and control rats (P &gt; 0.05). These findings show that increased sensitization of testicular cells to insulin signalling mediated by the PPAR γ agonist pioglitazone restores testicular histomorphometry and improves semen quality in diabetic rats, despite the weak effect of this drug on blood glucose. Conclusion: This suggests a role for insulin sensitizers and improved insulin signalling in the amelioration of testicular lesions and regulation of spermatogenesis in the diabetic state; and lends support for insulin at promoting testicular functions.

PPAR-γ agonist pioglitazone improves semen quality and testicular histomorphometrics with partial reversal of hyperglycaemia in alloxan-induced diabetic rats

Objective: Chronic hyperglycaemia is associated with subfertility and infertility in male diabetic subjects, possible as a result of perturbed testicular antioxidant defence system. Recent evidence however shows that some testicular cells produce insulin and this raises the possibility of a role for this hormone in spermatogenesis. Moreover, isoforms of the nuclear peroxisome proliferator activated receptor (PPAR) have been detected in testicular tissue, suggesting a role for insulin sensitisation in testicular function. In this study, we report the ameliorative effects of the PPAR γ agonist pioglitazone on testicular morphologic aberrations and impaired semen profile induced by diabetes in rats treated with intraperitoneal alloxan (150 mg/kg). Setting: This is an in vivo experimental study. Methods: Diabetic rats were randomized to receive oral pioglitazone at 10 or 30 mg/kg once daily for 6 weeks, after which rats were euthanised under anaesthesia. Caudal epididymal sperm was analysed for count, motility and morphology, and testosterone levels were measured in testicular homogenates by the enzyme immunoassay technique. Portions of the testes were fixed and processed by the H&E and Gordon and Sweet techniques. Blood glucose was estimated weekly during oral pioglitazone treatment using the glucometer. Results: Weekly blood glucose measurement showed weak effect of pioglitazone on hyperglycaemia in our model. Nonetheless, findings from caudal epididymal sperm analysis showed significantly high sperm density in the pioglitazone-treated rats at the two doses tested (P < 0.05 compared with diabetic controls). Moreover, pioglitazone at 10 mg/kg improved sperm motility and lowered the percentages of deformed and dead sperm cells in the treated diabetic rats. Haematoxylin and eosin (H&E) sections of the testes showed significantly reduced seminiferous tubule diameter and lumen size, as well as diminished height of the germinal epithelium in the untreated diabetic rats, but not in rats on pioglitazone intervention. Furthermore, Gordon and Sweet section of the testes showed thickened interstitial compartment only in the untreated diabetic group. Meanwhile, interstitial cells of Leydig appeared undisturbed in testicular sections of all the groups, while intratesticular testosterone levels were not significantly different between the treated and control rats (P > 0.05). Conclusion: These findings show that increased sensitivity of testicular cells to insulin mediated by the PPAR γ agonist pioglitazone restores testicular histomorphometry and improves semen quality in diabetic rats, despite the weak effect of this glitazone on blood glucose. This suggests a role for insulin sensitizers and improved insulin signalling in the amelioration of testicular lesions and regulation of spermatogenesis in the diabetic state, and lends support for insulin at promoting testicular functions.

Some Adverse Effects of Used Engine Oil (Common Waste Pollutant) On Reproduction of Male Sprague Dawley Rats.

AIM:Used oil is contaminated not only with heavy metals but also with polycyclic aromatic hydrocarbons (PAHs) that are insignificant in the unused oil. In our study we determined possible reproductive effects of used engine oil on male rats.MATERIAL AND METHODS:Twenty eight male Wistar rats were used for the study. The rats had average weight of 181.5 ± 10 g, animal feeds and portable water was provided ad-libitum. The rats were assigned to 4 groups (n = 7) including control. The treated groups orally received 0.1 ml/rat, 0.2 ml/rat and 0.4 ml/rat of the used engine oil every other day for 28 days using oral canulla. The spermatozoa were collected from epididymis for sperm analysis and testes were removed and preserved in Bouin’s fluid for routine histological analysis.RESULTS:Our results showed that there was progressive weight increase among the control group of rats that received distilled water. Meanwhile, rats that received 0.4 ml/rat of the used engine oil showed significant (P < 0.05) weight loss in second and third week of administration while rats that received 0.2 ml/rat and 0.1 ml/rat of the used engine oil showed non-significant (P > 0.05) weight reduction. The spermatozoa number was decreased with significance (P < 0.05) at 0.2 ml/rat (2.38 ± 0.29) and 0.4 ml/rat (1.98 ± 0.08) when compared with the control (5.00 ± 0.89). However, the percentage of motile sperms was reduced significantly (P <0.05) at 0.2 ml/rat (52.86 ± 3.59) and 0.4 ml/rat (45.71 ± 2.94) except at 0.1 ml/rat where the reduction (64.00 ± 7.5) was not significant (P> 0.05). The percentage of head deformity been 41.43 ± 2.61 and 42.00 ± 3.74 at 0.2 ml/rat and 0.4 ml/rat respectively, also significant increase of middle piece deformity was observed only at 0.1 ml/rat (45.71 ± 2.02) while tail deformity significantly decreased (15.71 ± 2.02, 20.00 ± 4.36 and 20.00 ± 4.47) when compared with the control (30.00 ± 1.29). The testicular seminiferous tubules were slightly degenerated with absence of Lumen. The germinal cell layer consisting of necrosis of spermatogonia and interstitial (Leydig) cells with affected Sertoli cells at different maturation stages.CONCLUSION:Hence, it can be said that there is a negative relation between used engine oil and male reproductive parameters. And it can be concluded that used engine oil should be prevented from leaking, spilling or improperly discarded as through medium it may enter storm water runoff and eventually affect the environmental health receiving water bodies.

Moringa oleifera extract enhances sexual performance in stressed rats.

Aphrodisiacs are required to improve male sexual function under stressful conditions. Due to the effects of oxidative stress and dopamine on male sexual function, we hypothesized that Moringa oleifera leaves might improve male sexual dysfunction induced by stress. Therefore, the effects on various factors playing important roles in male sexual behavior, such as antioxidant effects, the suppression of monoamine and phosphodiesterase type 5 (PDE-5) activities, serum testosterone and corticosterone levels, and histomorphological changes in the testes, of a hydroethanolic extract of M. oleifera leaves were investigated. Various doses of extract including 10, 50, and 250 mg/kg body weight (BW) were given orally to male Wistar rats before exposure to 12 h-immobilization stress for 7 d. The results demonstrated that the extract showed both antioxidant and monoamine oxidase type B (MAO-B) suppression activities. At 7 d of treatment, the low dose of extract improved sexual performance in stress-exposed rats by decreasing intromission latency and increasing intromission frequency. It also suppressed PDE-5 activity, decreased serum corticosterone level, but increased serum testosterone, numbers of interstitial cells of Leydig and spermatozoa. The increased numbers of interstitial cells of Leydig and spermatozoa might have been due to the antioxidant effect of the extract. The increased sexual performance during the intromission phase might have been due to the suppression of MAO-B and PDE-5 activities and increased testosterone. Therefore, M. oleifera is a potential aphrodisiac, but further research concerning the precise underlying mechanisms is still needed.

A Sertoli cell-specific connexin43 knockout leads to altered interstitial connexin expression and increased Leydig cell numbers.

The Sertoli cell (SC)-specific knockout (KO) of connexin43 (Cx43) results in spermatogenic arrest at the level of spermatogonia and/or SC-only syndrome. Histology of the interstitial compartment suggests Leydig cell (LC) hyperplasia. Our aim has been to investigate possible effects of the SC-specific KO of Cx43 (SCCx43KO) on interstitial LC. We therefore counted LC via the optical dissector method (per microliter of testicular tissue and per testis) and found LC to be significantly increased in SCCx43KO(-/-) compared with wild-type mice. Semiquantitative western blot together with Cx43 and 3β-hydroxysteroid dehydrogenase immunohistochemistry showed that Cx43 protein was significantly reduced and barely detectable in LC in adult SCCx43KO(-/-) mice. This reduction of Cx43 protein was accompanied by a reduction of Cx43 mRNA as analyzed by laser-assisted microdissection of interstitial cells and subsequent quantitative real-time polymerase chain reaction (PCR). Interestingly, Cx45, another recently detected connexin in LC, was also downregulated. Preliminary qualitative data of LC differentiation markers (Thb2, Hsd3b6) and a steroidogenic marker (Hsd17b3) obtained by reverse transcription plus PCR revealed no obvious differences. Thus, the loss of Cx43 in SC also provokes the downregulation of connexins in interstitial LC at the transcriptional and translational levels. Moreover, SCCx43KO leads to alterations in LC numbers. Despite these alterations, steroidogenesis seems not to be impaired. Further studies, including ultrastructural analysis of the tissue as well as quantitative examination of additional LC markers and testosterone, and functional in vitro experiments, should provide more information about LC differentiation and function in SCCx43KO(-/-) mice.

IMPACT OF 4-NONYLPHENOL ON TESTOSTERONE PRODUCTION IN MICE LEYDIG CELLS

Nonylphenol (NP) is an important environmental toxicant and potential endocrine disrupting chemical. It is also the primary final biodegradation product of nonylphenol polyethoxylate, a non-ionic surfactant that is frequently incorporated into detergent formulation. Several studies have reported adverse effects of NP on the male reproductive system, including hormonal changes and testicular abnormalities. However, its mechanisms are not clearly understood. In the present study, experiments were performed to examine the effects of 4-nonylphenol on the testosterone production in mice Leydig cells. Interstitial (Leydig) cells were cultured with addition of 0.04, 0.2, 1.0, 2.5 and 5 μg/mL of 4-nonylphenol and compared with control. Concentrations of testosterone in the media were determined using enzyme linked immunosorbent assay (ELISA). Hormone production significantly (P

Histological Changes of Selected Westar Rat Tissues Following the Ingestion of Tartrazine With Special Emphasis on the Protective Effect of Royal Jelly and Cod Liveroil

The present investigation was carried out on thirty six mature Westar rats from laboratory animal unit in the faculty of veterinary medicine, Zagazig University, Egypt. The animals were divided into six groups of six animals for each. Group I was considered as a control group. Group II was treated with tartrazine only in a dose 500 mg/kg body weight by oral gavage for 30 day. Group III was treated with the same dose of tartrazine with 300 mg/kg body weight Royal jelly. Group IV was treated with tartrazine and Code liver oil in a dose 0.4 mg/kg body weight. While the last two groups, V and VI were treated with Code liver oil and Royal jelly separately by the above mentioned doses for 30 days. At the end of experiment, cervical dislocation of rats and for the histological studies, the liver, kidney, testis and brain tissues in addition to stomach were separated and small pieces from them were taken, fixed in neutral buffered formalin 10%, dehydrated, cleared and processed for paraffin blocks and 5 micron sections were obtained, mounted on a glass slides and stained histologically with Hematoxylin and Eosin (H&E) and Blue Masson’s Trichrome. Groups I, V, VI showed normal histological structure of all selected tissues without any abnormalities. Group II showed severe steatosis, diffuse degeneration, necrosis of hepatic tissues, fibrous tissue proliferation with anti-inflammatory cells infiltration in the portal areas and moderate disorganization of hepatic cords. Furthermore, hyperplasia of the interstitial connective tissue and Leydig cells with marked vacuolations were recognized. Moreover, slightly vacuolations in brain tissues were recognized specially within the white matter as well as degenerative changes in stomach mucosa. And also, hyaline degeneration in renal tubules and vacuolations of glomeruli were observed. The present investigation concluded that the serious effect of tartrazine toxicity was more pronounced in liver then kidney, stomach, testis and brain tissues respectively. Moreover, the curative protective effect of both royal jelly and cod liver oil are non-significant against tartrazine toxicity.

Chemical Sterilization of Dogs using Single Bilateral Intra-testicular Injection of Calcium Chloride or Clove Oil

The current study was designed to evaluate the efficacy of single bilateral intra-testicular injection of calcium chloride or clove oil to induce chemical sterilization in ten dogs divided into two groups each of five animals. Chemical sterilization was evaluated by clinical signs, hormonal analysis and histopathological examination. Testosterone level was decreased significantly after injection of both agents with complete degeneration of the interstitial leydig cells and seminiferous tubules while cortisol level was increased during the 1st week after calcium chloride injection. The single bilateral intra-testicular injection of calcium chloride or clove oil proved to be accepted as a suitable alternative to the surgical sterilization of dogs; however clove oil was superior to calcium chloride as it induced less pain and irritation to the dogs.

Molecular adaptations of testosterone-producing Leydig cells during systemic in vivo blockade of the androgen receptor

This study systematically evaluates the effects of androgen receptor (AR) blockade on molecular events in Leydig cells. Results showed that intramuscular administration of testosterone-enanthate, at clinically relevant dose, decreased testosterone in interstitial fluid and Leydig cells from adult rats. AR-blocker (Androcur) prevented this effect and testosterone-reduced Leydig cells steroidogenic capacity/activity. Testosterone-reduced expression of some steroidogenic enzymes/proteins (Tspo,StAR,Hsd3b1/2) and transcription factors (Nur77,Gata4,Dax1) was completely abrogated, while decreased expression of Star,Cyp11a1,Cyp17a1,Hsd17b4,Creb1a was partially prevented. In the same cells, increased expression of Hsd3b5/HSD3B and Ar/AR was abolished. Androcur-treatment abolished testosterone-reduced cAMP, coupled with a changed expressional milieu of cAMP signaling elements. Results from in vitro experiments suggest that some of these effects are testosterone-AR dependent, while others could be due to disturbed LH and/or other signals. Presented data provide new molecular insight into Leydig cells function and are important in terms of human reproductive health and the wide-spread use of Androcur as well as use/abuse of testosterone-enanthate.

Black tea extract ameliorates indomethacin induced changes in testicular histopathology of albino rats

Context: Reactive oxygen species and lipid peroxidation play a role in the pathogenesis induced by the non-steroidal antiinflammatory drug indomethacin. Aims: This study was aimed to assess the simultaneous effect of black tea extract (BTE) on indomethacin induced alteration in testicular pathology. Materials and Methods: Adult rats were divided into four groups (n = 6/group); Group I (control); Group II (indomethacin, 5 mg/kg b.wt.; i.p.); Group III (BTE, 2.5 g tea leaf/dL of water, i.e., 2.5% of aqueous BTE, orally) and Group IV (indomethacin + BTE). Serum vitamin E, sperm count, motility, and histopathology of testes were evaluated. Statistical Analysis: Statistical comparisons were performed using the one-way ANOVA, followed by post-hoc t-test. Results: Indomethacin induced rats showed a significant decrease in testicular weight, sperm count, sperm motility, and serum vitamin E concentrations. Histopathology of the testes revealed tortuous seminiferous tubules, loss of spermatogenesis process, marked decrease in the number of interstitial cells of Leydig and been replaced by many foci of congestion, edema, necrosis, and interstitial fibrosis. Rats simultaneously treated with BTE and indomethacin showed improvement of testicular weight, sperm count, sperm motility, and serum vitamin E concentrations when compared to indomethacin alone treated rats. Similarly, histopathology of rats treated simultaneously with indomethacin and BTE showed near normal testicular architecture when compared to indomethacin alone treated rats. Conclusion: The result suggests that BTEs might have potential beneficial effect to combat against indomethacin induced testicular damage in rats. Key words: Black tea extract, indomethacin, testicular damage, vitamin E