Leydig Cell Number Research Articles

Ameliorating effects of curcumin-loaded superparamagnetic iron oxide nanoparticles (SPIONs) on the mouse testis exposed to the transient hyperthermia: A molecular and stereological study

BackgroundTesticular hyperthermia can have negative effects on male fertility. Despite reported therapeutic benefits of curcumin, several factors often limit its application such as low water solubility and instable structure. Curcumin-loaded superparamagnetic iron oxide nanoparticles (SPIONs) were designed to solve its limitation of use. In the present study, we evaluated the effect of curcumin-loaded SPIONs on transient testicular hyperthermia in mouse. Materials and methodA total of 18 adult male NMRI mice were divided into three groups (n = 6): I. Controls (Cont), II. Scrotal hyperthermia (Hyp), III. Scrotal hyperthermia + curcumin-loaded iron particles (240 μL) (Hyp + Cur). After seventy days, the animals were sacrificed and used for further molecular and stereological evaluations. ResultsSperm count, motility and viability significantly decreased in group hyp as compared to cont group. Furthermore, Sperm DNA fragmentation and cell apoptosis in testes increased remarkably in group hyp, compared with group cont. Stereological study showed a reduction in number of spermatogenic and Leydig cells, as well as reduced weight and volume of testes in hyp group. Degenerative appearance of testes exposed to hyperthermia was also observed. In addition, higher mRNA expression of inflammatory cytokines (IL1-α, IL6, and TNF-α) was detected in group hyp compared to cont group. However, curcumin-loaded SPIONs alleviated all of the pathologic changes in the Hyp + Cur group compared to the hyp group. ConclusionHere, we used nanoparticle form of curcumin in testicular hyperthermia model and showed its ameliorating effects on testes damages caused by heat stress, which can be an appropriate method to overcome the problems that limit curcumin application in cases with increased intra testicular temperature.

Stereological assessment of the effects of vitamin D deficiency on the rat testis

BackgroundAccumulating evidence suggests that low vitamin D status may affect male gonadal structure. This study was undertaken to reveal whether vitamin D-deficient rats have demonstrable changes in the quantitative histomorphometric properties of the testis.MethodsIn the present investigation, adult male Sprague-Dawley rats were divided into four groups and received: group 1) conventional diet; group 2) vitamin D-deficient diet; group 3) vitamin D-deficient diet and paricalcitol and group 4) conventional diet plus paricalcitol. After 3 months, serum levels of vitamin D metabolites, Ca, P, LH, FSH, testosterone, and epididymal sperm quality were evaluated. Moreover, the morphometric characteristics of testis were assessed via stereological methods.ResultsRats fed a vitamin D-deficient diet (groups 2 and 3) were normocalcemic and had 25-hydroxyvitamin D3 level below 10 ng/mL. A significant reduction in serum testosterone and comparable gonadotropin levels were seen in vitamin D-deficient groups compared to controls. The concentration, morphology, and motility of sperm cells were profoundly disturbed in animals raised on the vitamin D-deficient diet. There was a significant decline in the population of different germ cells, the volume of interstitial tissue and germinal epithelium in group 2 and 3 rats, which were placed on the vitamin D-deficient diet. No appreciable difference in the estimates of the Leydig or Sertoli cell numbers were observed between groups.ConclusionsThe depletion of vitamin D stores and induction of moderate grades of vitamin D deficiency by dietary measures led to remarkable impairment of spermatogenesis and microscopic architecture of rat testis. These findings can be attributed, at least in part, to decreased androgen production.

Age-related changes in human Leydig cell status.

What are the consequences of ageing on human Leydig cell number and hormonal function? Leydig cell number significantly decreases in parallel with INSL3 expression and Sertoli cell number in aged men, yet the in vitro Leydig cell androgenic potential does not appear to be compromised by advancing age. There is extensive evidence that ageing is accompanied by decline in serum testosterone levels, a general involution of testis morphology and reduced spermatogenic function. A few studies have previously addressed single features of the human aged testis phenotype one at a time, but mostly in tissue from patients with prostate cancer. This comprehensive study examined testis morphology, Leydig cell and Sertoli cell number, steroidogenic enzyme expression, INSL3 expression and androgen secretion by testicular fragments in vitro. The majority of these endpoints were concomitantly evaluated in the same individuals that all displayed complete spermatogenesis. Testis biopsies were obtained from 15 heart beating organ donors (age range: 19-85 years) and 24 patients (age range: 19-45 years) with complete spermatogenesis. Leydig cells and Sertoli cells were counted following identification by immunohistochemical staining of specific cell markers. Gene expression analysis of INSL3 and steroidogenic enzymes was carried out by qRT-PCR. Secretion of 17-OH-progesterone, dehydroepiandrosterone, androstenedione and testosterone by in vitro cultured testis fragments was measured by LC-MS/MS. All endpoints were analysed in relation to age. Increasing age was negatively associated with Leydig cell number (R = -0.49; P < 0.01) and concomitantly with the Sertoli cell population size (R= -0.55; P < 0.001). A positive correlation (R = 0.57; P < 0.001) between Sertoli cell and Leydig cell numbers was detected at all ages, indicating that somatic cell attrition is a relevant cellular manifestation of human testis status during ageing. INSL3 mRNA expression (R= -0.52; P < 0.05) changed in parallel with Leydig cell number and age. Importantly, steroidogenic capacity of Leydig cells in cultured testis tissue fragments from young and old donors did not differ. Consistently, age did not influence the mRNA expression of steroidogenic enzymes. The described changes in Leydig cell phenotype with ageing are strengthened by the fact that the different age-related effects were mostly evaluated in tissue from the same men. In vitro androgen production analysis could not be correlated with in vivo hormone values of the organ donors. In addition, the number of samples was relatively small and there was scarce information about the concomitant presence of potential confounding variables. This study provides a novel insight into the effects of ageing on human Leydig cell status. The correlation between Leydig cell number and Sertoli cell number at any age implies a connection between these two cell types, which may be of particular relevance in understanding male reproductive disorders in the elderly. However aged Leydig cells do not lose their in vitro ability to produce androgens. Our data have implications in the understanding of the physiological role and regulation of intratesticular sex steroid levels during the complex process of ageing in humans. This work was supported by grants from Prin 2010 and 2017. The authors have no conflicts of interest. N/A.

Neurotrophin-3 stimulates stem Leydig cell proliferation during regeneration in rats.

Neurotrophin‐3 (NT‐3) acts as an important growth factor to stimulate and control tissue development. The NT‐3 receptor, TRKC, is expressed in rat testis. Its function in regulation of stem Leydig cell development and its underlying mechanism remain unknown. Here, we reported the role of NT‐3 to regulate stem Leydig cell development in vivo and in vitro. Ethane dimethane sulphonate was used to kill all Leydig cells in adult testis, and NT‐3 (10 and 100 ng/testis) was injected intratesticularly from the 14th day after ethane dimethane sulphonate injection for 14 days. NT‐3 significantly reduced serum testosterone levels at doses of 10 and 100 ng/testis without affecting serum luteinizing hormone and follicle‐stimulating hormone levels. NT‐3 increased CYP11A1‐positive Leydig cell number at 100 ng/testis and lowered Leydig cell size and cytoplasmic size at doses of 10 and 100 ng/testis. After adjustment by the Leydig cell number, NT‐3 significantly down‐regulated the expression of Leydig cell genes (Lhcgr, Scarb1, Star, Cyp11a1, Hsd3b1, Cyp17a1, Hsd17b3, Hsd11b1, Insl3, Trkc and Nr5a1) and the proteins. NT‐3 increased the phosphorylation of AKT1 and mTOR, decreased the phosphorylation of 4EBP, thereby increasing ATP5O. In vitro study showed that NT‐3 dose‐dependently stimulated EdU incorporation into stem Leydig cells and inhibited stem Leydig cell differentiation into Leydig cells, thus leading to lower medium testosterone levels and lower expression of Lhcgr, Scarb1, Trkc and Nr5a1 and their protein levels. NT‐3 antagonist Celitinib can antagonize NT‐3 action in vitro. In conclusion, the present study demonstrates that NT‐3 stimulates stem Leydig cell proliferation but blocks the differentiation via TRKC receptor.

Exposure to di-n-octyl phthalate during puberty induces hypergonadotropic hypogonadism caused by Leydig cell hyperplasia but reduced steroidogenic function in male rats

Humans are exposed to phthalates ubiquitously, which may threaten health. However, whether di-n-octyl phthalate can prevent pubertal sexual maturity is still elusive. In this study, male Sprague Dawley rats (age 35 days) were treated daily by gavage with 0, 10, 100, and 1000 mg/kg body weight of di-n-octyl phthalate from day 35 to day 49 after birth. Di-n-octyl phthalate significantly reduced serum testosterone levels at doses of 100 and 1000 mg/kg, but increased serum luteinizing hormone levels of 1000 mg/kg and decreased testosterone/luteinizing hormone ratio at ≥10 mg/kg, without affecting serum follicle-stimulating hormone levels. Di-n-octyl phthalate significantly induced Leydig cell hyperplasia (increased number of CYP11A1-positive Leydig cells) at 100 and 1000 mg/kg. Di-n-octyl phthalate down-regulates the gene expression of Cyp11a1, Hsd3b1 and Insl3 in individual Leydig cells. Di-n-octyl phthalate can also reduce the number of sperm in the epididymis. Di-n-octyl phthalate increased phosphorylated AKT1/AKT2 without affecting their total proteins, but increased the total protein and phosphorylated protein of ERK1/2 and GSK-3β. Primary immature Leydig cells isolated from 35-day-old rats were treated with 0–50 μM di-n-octyl phthalate for 3 h. This phthalate inhibited androgen production under basal, LH-stimulated, and cAMP-stimulated conditions by 5 and 50 μM in vitro via down-regulating Cyp11a1 expression but up-regulating Srd5a1 expression in vitro. In conclusion, di-n-octyl phthalate induces hypergonadotropic hypogonadism caused by Leydig cell hyperplasia but reduced steroidogenic function and prevents sperm production.

INSL3: A Marker of Leydig Cell Function and Testis-Bone-Skeletal Muscle Network.

This article reviews the role of INSL3 as biomarker of Leydig cell function and its systemic action in testis-bone-skeletal muscle crosstalk in adult men. Insulin-like factor 3 (INSL3) is a peptide hormone secreted constitutively in a differentiation-dependent mode by testicular Leydig cells. Besides the role for the testicular descent, this hormone has endocrine anabolic functions on the bone-skeletal muscle unit. INSL3 levels are low in many conditions of undifferentiated or altered Leydig cell status, however the potential clinical utility of INSL3 measurement is not yet well defined. INSL3 levels are modulated by the long-term cytotropic effect of the hypothalamicpituitary- gonadal axis, unlike testosterone that is acutely sensitive to the stimulus by luteinizing hormone (LH). INSL3 directly depends on the number and differentiation state of Leydig cells and therefore it represents the ideal marker of Leydig cell function. This hormone is more sensitive than testosterone to Leydig cell impairment, and the reduction of INSL3 in adult men can precociously detect an endocrine testicular dysfunction. Low INSL3 levels could cause or contribute to some symptoms and signs of male hypogonadism, above all sarcopenia and osteoporosis. The measurement provided suggested that the measurement of INSL3 levels should be considered in the clinical management of male hypogonadism and in the evaluation of testicular endocrine function. The monitoring of INSL3 levels could allow an early detection of Leydig cell damage, even when testosterone levels are still in the normal range.

Ethanol exposure during the brain growth spurt period increases ethanol‐induced aggressive behavior in adolescent male mice

Ethanol exposure during development is associated with deficient social behavior, such as aggressive behavior, and ethanol consumption is associated with violent crimes, thus raising the possibility that individuals with fetal alcohol spectrum disorder may exhibit exacerbated social deficits in response to ethanol exposure. The present study evaluated the effects of ethanol exposure during the brain growth spurt period (i.e., a critical time period during which ethanol's effects are augmented) on aggressive behavior and ethanol-induced aggression during adolescence. From postnatal Day 2 (PD2) to PD8, Swiss mice received either ethanol (5g/kg, i.p.) or saline on alternate days. On PD39, aggressive behavior was assessed using the resident-intruder paradigm in male mice, and social dominance was investigated using the tube dominance test in both males and females. Testis structure and testosterone levels were evaluated in male mice. Early ethanol exposure increased the gonadosomatic index and the number of Leydig cells. The thickness of the seminiferous tube decreased. No difference in testosterone levels was found. The ethanol-exposed resident mice exhibited increased number and duration of aggressive episodes only when challenged with a low ethanol dose (1g/kg) before confrontation. Female mice early-exposed to ethanol won more confrontations in the tube dominance test. The present findings suggest a critical brain growth spurt period that is susceptible to ethanol-induced alterations of social dominance behavior in females. Although basal levels of aggression were unaffected, early ethanol exposure resulted in greater susceptibility to ethanol-induced aggression in adolescent male mice.

Effect of Zirconium oxide nanoparticle on serum level of testosterone and spermatogenesis in the rat: An experimental study.

Background Zirconium nanoparticles are used as health agents, pharmaceutical carriers, and in dental and orthopedic implants.ObjectiveThis studyaimed to investigate the effects of Zirconium oxide nanoparticles on the process of spermatogenesis in rat.Materials and MethodsIn this experimental study, 32 male Wistar rats (150-200 gr), with range of age 2.5 to 3 months were used and divided into four groups of eight per each. The control group received 0.5 ml of distilled water and the three experimental groups received 50, 200, and 400 ppm doses of Zirconium oxide nanoparticles solution over a 30-day period, respectively. At the end of the experiment, tissue sections were taken from the testis and stained with hematoxylin-eosin. Serum concentration of testosterone was measured by enzyme-linked immunosorbent assay.Results In the experimental group receiving 400 ppm Zirconium oxide nanoparticles, the number of Spermatogonia cells (p 0.01), Spermatocytes (p 0.01), Spermatids (p 0.001), and sertoli and Leydig cells (p 0.05) showed a significant decrease compared to the control group. Serum testosterone concentration did not change significantly in all experimental groups receiving Zirconium oxide nanoparticles compared to the control group. Experimental group received 400 ppm Zirconium oxide nanoparticles shrinkage of seminal tubules and reduced lumen space compared to control group.Conclusion Zirconium oxide nanoparticles are likely to damage the testes by increasing Reactive oxygen species production and free radicals.

Effects of gonadotropins on testis cell subpopulations of newly hatched chicks treated during embryonic development

The aim of the present study was to evaluate histological and stereological changes, as well as the variations in the number and size of cells from diverse cell subpopulations in testes of newly hatched chicks treated with follicle stimulating hormone (FSH) and luteinizing hormone (LH) during embryonic development. Stereological results indicated that in FSH-treated chicks total volume of the tubular compartment constitutes most of the testis. In contrast, the total volume of interstitial tissue constitutes most of the testis of LH-treated chicks. Results indicate that the number of germ and Sertoli cells increases as a result of FSH and LH treatment, but in FSH-treated testis, Sertoli cells were the most numerous cell type in seminiferous tubules; whereas germ cells were the most numerous cell type in testis of LH-treated chicks. Results also indicate there was a larger total volume of Leydig cells in the testes of FSH- and LH-treated chicks. The larger volume of Leydig cells in FSH-treated chicks is due to a larger cellular volume of these cells, and not due to the number, which remains constant. In contrast, in testes of LH-treated chicks, there is a larger number and volume of Leydig cells. These results indicate the testes of chick embryos respond to FSH and LH treatment, with there being modifications in the seminiferous tubules and interstitial tissue, but these changes differ markedly, indicating that FSH and LH have differential effects on chick testes.

Effects of Icariin on Histomorphometric Changes of Testis and Prostate Induced by Acrylamide in Mice

Background and Aims: This study aimed to observe the effect of Icariin on histomorphometric changes of testis and prostate induced by Acrylamide. Materials and Methods: Male mice were divided into four groups (n=8): A is the control group and does not get any treatment, B is the sham group and only received drinking water. C group received Acrylamide 10 mg/kg. D group received Acrylamide 15 mg/kg+1.5 mg/kg of Icariin. Histological changes in testis and prostate were examined using stereological methods. Results: Results showed decreases in testis weight of the group treated by (p≤0.01) and the group cured by Acrylamide +Icariin group (p≤0.05). The total volume of testis showed a reduction in the Acrylamide group compared to other groups (p≤0.05). The total number of spermatogonia and spermatocyte cells in the Acrylamide group showed a decrease in comparison with the other groups (p≤0.05). The total number of spermatid cells in the Acrylamide group indicated a significant reduction in comparison with the control and sham group (p≤0.05). The total number of sertoli cells in the Acrylamide group showed a reduction when the number of leydig cells in the Acrylamide group showed a significant decrease in comparison with the control, sham, and Acrylamide+Icariin groups (p≤0.05). The mean Johnsen score was decreased in the Acrylamide treated group compared to control, sham, and Acrylamide+Icariin groups (p≤0.05). Testosterone concentration in the Acrylamide group showed a reduction in comparison with control, sham, and Acrylamide+Icariin groups (p≤0.05). Conclusion: Results demonstrated that Acrylamide altered the structure of the testis, prostate gland, and spermatogenesis stage, and Icariin treatment improved these histopathological changes.

Pregabalin administration and withdrawal affect testicular structure and functions in rats.

The aim of this prospective experimental study was to investigate the effects of pregabalin (PG) administration and withdrawal on testicular structures and functions in rats. A total of 24 male Wistar rats were divided into two groups (n=12 each): a control group received normal saline, and PG-treated group received 62mgkg-1 day-1 PG for 2months. Half the animals of each group were sacrificed for the collection of blood and testicular samples. The remaining animals were bred for another 2months without treatment before collection of blood and testicular samples. PG administration decreased testosterone and increased luteinising hormone (LH) and follicle-stimulating hormone (FSH) levels versus controls. PG withdrawal led to a decrease in both FSH and LH and an increase in testosterone levels versus saline withdrawal. Compared to controls, PG administration caused degeneration of seminiferous tubules and decreased the number of spermatogenic but increased the number of Leydig cells. After PG withdrawal, these cells showed a rebound reverse. Reduced glutathione levels increased with PG administration while PG withdrawal increased malondialdehyde levels. Conclusion: PG administration affected testicular morphometry, gonadotrophic and sex hormones; however, there was a rebound reversal in all these parameters and a significant oxidative stress in PG withdrawal.

Testicular biometry, ultrastructure, cellular cytometry and hormonal profile in relation to age in Egyptian buffaloes.

This study was conducted on a number of (30) buffaloes in Qalubia. The age ranged from 12 to 36 month. There were divided into two groups: group (I) and group (II), their ages ranged from <18 month and ≥18 till 36 months, respectively. Testosterone levels in the blood, testicular biometry by the optical and the electronic microscope, and finally flow cytometry of the testicular cells were estimated. it was concluded that: The testosterone hormone level in the blood in group (I) and in group (II) was 1.9 ± 0.48 and 1.07 ± 0.35 ng/ ml serum prospectively, weight, length and width of the testicle in group (I) was 75± 13.8 gm., 7.3 ± 0.48cm and 4.22 ± 0.31 cm, respectively, and in group (II) was 118.38 ± 11.29 gm., 8.12 ± o.63 cm, and 5.0 ± 0.23 cm, respectively. The diameter of the seminiferous tubules in group (I) was 99.90 ± 17.4µm, while in group (II) was 104.6 ± 15.82 µm. The number of Leydig cells in group (I) was 40.14 ± 15.52, and in group (II) was 49.16 ± 27.99. The lining epithelium thickness of the seminiferous tubules in group (I) was 26.06 ± 5.51 µm, and in group (II) was 27.64 ±5.52 µm, while degree of cell degeneration in group (I) was 6.12±0.79 and in group (II) was 4.62±0.66. By using flow cytometer there was difference between 2 age group in the testicular cell and apoptosis. An electron micrograph showing difference in the testicular cells’ ultrastructure

Fetal Leydig cells in buffalo testis: Light and electron microscopic study

The present study was conducted on testis of 20 buffalo fetuses ranging from 2.5 cm to 20 cm curved crown rump length (CVRL). The tissues were processed for light and transmission electron microscopic study. The intertubular tissue was filled with primitive mesenchymal cells, developing Leydig cells and blood vessels. At 7.5 cm CVRL, some of the interstitial mesenchymal cells started developing into foetal Leydig cells and were androgen positive. These cells were characterized by their polygonal shape, eosinophilic cytoplasm and darkly stained spherical nucleus with prominent nucleoli at 9.5 cm CVRL. At 15.5 cm CVRL, the fetal Leydig cells were present in groups around the interstitial blood vessels, which increased in number and size at 20 cm CVRL and decreased afterwards. By TEM, three types of Leydig cells were observed at 14.5 cm CVRL as stem Leydig cells (Leydig cell A), progenitor Leydig cells (Leydig cell B) and immature Leydig cells (Leydig cell C). Leydig cell A had elongated nucleus with a very thin rim of cytoplasm. Leydig cell B had oval nucleus with comparatively larger amount of cytoplasm. Leydig cell C had adult Leydig cell like features and had round to oval nucleus with lager amount of cytoplasm. The cytoplasm of these cells contained mitochondria with elongated cristae, Golgi apparatus and very few lipid inclusions. The number and size of Leydig cells increased during early prenatal life.

ASTAXANTHIN MENGHAMBAT PENURUNAN JUMLAH SEL LEYDIG DAN KADAR TESTOSTERON PADA TIKUS WISTAR JANTAN YANG DIPAPAR DENGAN ASAP ROKOK

Background. The exposure of cigarette smoke impacts negatively on Leydig cell count and testosterone level. Astaxanthin is known for its ability to neutralize free radicals far more potent than any other kind of antioxidant.&#x0D; Research objectives. This research aims to prove the effect of Astaxanthin in inhibiting decrease of Leydig cell count and testosterone level in male Wistar rats exposed with cigarette smoke.&#x0D; Methodology. The posttest only control group study design was conducted on 36 male Wistar rats, 12-16 weeks in age, with 200-210 grams body weight. Samples were randomly divided into two groups, consisting of a control group exposed with cigarette smoke and 0.5 ml of aquadest and a study group exposed with cigarette smoke and 0.1 mg of astaxanthin/200 gr BW daily for 30 days. On day 31, blood samples were taken to measure the testosterone level. Both testes were taken for Leydig cells count assessment. Comparative analysis was done to see any significant difference between the study and control group.&#x0D; Research results. The results show that the mean number of Leydig cells and testosterone levels in the study group was significantly higher than the control group (p&lt;0.01).&#x0D; Conclusion. Oral astaxanthin administration inhibited the decrease of Leydig cell count and testosterone level in male Wistar rats exposed with cigarette smoke.&#x0D; Key words: Astaxanthin, Testosterone, Leydig Cell, Andropause

Hypothyroidism induced by postnatal PTU (6-n-propyl-2-thiouracil) treatment decreases Sertoli cell number and spermatogenic efficiency in sexually mature pigs

Studies with 6-n-propyl-2-thiouracil (PTU) in laboratory rodents have shown that transient neonatal hypothyroidism leads to increased Sertoli cell (SC) number, testis size and sperm production. However, scarce and inconclusive data are available for farm animals. In the present study, Piau pigs received PTU in a gel capsule containing 8 mg/kg of body weight for 14 weeks starting from the first week of age, whereas control animals received only the vehicle. Blood samples were collected during the experimental period for hormonal evaluation in the serum. The animals were orchiectomized at adulthood and had their testes used for histomorphometric analysis. Indicating that the PTU concentration used was effective in promoting hypothyroidism, PTU-treated pigs showed a 30% lower body weight and reduced thyroxine levels (p < 0.05) during the treatment period. At adulthood, the body weight was similar in both groups but, surprisingly, PTU-treated pigs showed 30% lower testis weight (p < 0.05). In general, treated pigs presented increased follicle-stimulating hormone levels, whereas testosterone levels tended to be lower from 9 to 23 weeks of age. No significant differences were observed for estradiol, Leydig cell volume and number, tubular diameter, SC number per gram of testis, SC efficiency and meiotic index. However, seminiferous tubule occupancy, total tubular length, SC number per testis, and daily sperm production per testis and per gram of testis (DSP/g/T) were significantly lower (p < 0.05) in PTU-treated pigs. Therefore, in contrast to laboratory rodents, our results showed that SC proliferation and DSP/g/T (spermatogenic efficiency) in Piau pigs is diminished by postnatal PTU treatment.

Gestational exposure to tebuconazole affects the development of rat fetal Leydig cells

Tebuconazole is a triazole fungicide, used in agriculture to treat phytopathogenic fungi, and as a biocide, has been reported to be related to reproductive and developmental toxicity. The purpose of this study was to investigate the effect of tebuconazole exposure on rat fetal Leydig cells and fetal testis during pregnancy. Pregnant Sprague-Dawley rats were randomly divided into 4 groups, daily gavaged with corn oil (as a control), 25, 50, and 100 mg/kg body weight tebuconazole for 10 days (from the 12th day of pregnancy). Tebuconazole increased fetal serum testosterone and progesterone levels at a dose of 100 mg/kg. Exposure to 100 mg/kg tebuconazole significantly caused an increase in the number of fetal Leydig cells per testis without inducing cell aggregation. Tebuconazole up-regulated the expression of Star, Cyp11a1, Hsd17b3, and Fshr and their proteins. Further investigation found that tebuconazole caused increased phosphorylation of AKT1, ERK1/2, and mTOR, the level of BCL2, as well as the decrease of Beclin1, LC3B, and BAX, which may contribute to the fetal Leydig cell autophagy and proliferation. In conclusion, in utero exposure of tebuconazole causes the proliferation of fetal Leydig cells.

Cytological description of testicular cell populations in sexually mature cats with normal spermatogenesis.

In cats, assessment of the testicular function is mainly based on sperm evaluation. Whatever the technique used, the volume of collected sperm is often small, which may lead to technical difficulties to achieve the semen evaluation in routine practice. Fine-needle aspiration (FNA) biopsy of the testicular parenchyma is one of the other methods used to assess testicular function. The aim of this study was to explore the relevance of FNA in the assessment of testicular cells in sexually mature cats. Eighteen cats over one year of age were recruited among animals presented for surgical neutering. Semen was collected by electroejaculation before it was evaluated. FNA biopsies of the testicles were taken using a 21-gauge needle. After castration, histological analysis of the testes was performed. Semen evaluation and histological analysis showed no anomalies, which confirmed normal spermatogenesis in all the cats and allowed a proper interpretation of the cytological findings. The cells identified through cytological examination were spermatogonia (1.99±0.17%), primary spermatocytes (10.49±0.74%), round spermatids (34.80±1.57%), elongated spermatids (23.59±2.02%), spermatozoa (21.56±1.86%), Sertoli cells (7.53±1.23%) and Leydig cells (0.04±0.03%). However, spermatocytes II were not identified. This is due to the low proportions of these cells, related to their very short lifespan. Likewise, the very low number of Leydig cells observed is probably due to the damage caused during the aspiration stage. This study showed that fine-needle aspiration is an efficient method to describe cytologically normal testicular populations, a cornerstone for future research aimed to study abnormal spermatogenesis and to correlate it to cytological proportion of germ cells.

Gestational vinclozolin exposure suppresses fetal testis development in rats

Vinclozolin is a common dicarboximide fungicide used to protect crops from diseases. It is also an endocrine disruptor and is thought to be related to abnormalities of the reproductive tract. However, its mechanism of inducing abnormalities of the male reproductive tract is still unclear. The purpose of this study was to study the effect of gestational vinclozolin exposure on the development of rat fetal Leydig cells. Female pregnant Sprague-Dawley rats were exposed to vinclozolin (0, 25, 50, and 100 mg/kg body weight/day) by gavage from gestational day 14–21. Vinclozolin dose-dependently reduced serum testosterone levels at doses of 50 and 100 mg/kg and the anogenital distance at 100 mg/kg. RNA-seq, qPCR, and Western blotting showed that vinclozolin down-regulated the expression of Nr5a1, Sox9, Lhcgr, Cyp11a1, Hsd3b1, Hsd17b3, Amh, Pdgfa, and Dhh and their encoded proteins. Vinclozolin reduced the number of NR2F2-positive stem Leydig cells at a dose of 100 mg/kg and enhanced autophagy in the testes. In conclusion, vinclozolin disrupts reproductive tract development and testis development in male fetal rats via several pathways.

Effect of tamsulosin on testis histopathology and serum hormones in adult rats: Experimental study.

Background Tamsulosin is an inhibitory factor of alpha-adrenergic receptors that is used for relieving of the clinical symptoms and management of acute urinary retention.ObjectiveThe aim of this study was to evaluate the effects of tamsulosin on the endocrine axis and testicular tissue in adult male rats.Materials and MethodsIn this experimental study, 30 adult male Wistar rats (weighing 250-300 gr) were divided into three groups: 1) control (received distilled water), 2) experimental 1 (received 0.2 mg/kg/day tamsulosin) and 3) experimental 2 (received 0.4 mg/kg/day tamsulosin) through oral gavage for 28 days. Serum hormones level and testicular histopathology were evaluated at the end of the experiment.ResultsIn this study, the testicular weight decreased significantly in the experimental groups compared to the control group. A significant decrease was seen in testicular weight (p = 0.004) and the number of Leydig cells in tamsulosin-treated groups (p = 0.012). Tamsulosin improved the hormone profile in experimental groups. Also, higher dose of tamsulosin significantly changed the number of Leydig, spermatogonia cells, the thickness of germinal layer, and the diameter of the seminiferous tubules.Conclusion Results showed that using tamsulosin, possibly reduces the testosterone concentration through adrenergic axis system and in turn has destructive effects on proliferative activity of germ cells.

Di-n-hexyl phthalate causes Leydig cell hyperplasia in rats during puberty

Di-n-hexyl phthalate (DNHP) is commonly used as a plasticizer. However, whether DNHP influences Leydig cell development during puberty remains unexplored. In this study, DNHP (0, 10, 100, and 1000 mg/kg) was administered via gavage to 35-day-old male Sprague-Dawley rats for 21 days. Serum levels of testosterone, luteinizing hormone, follicle-stimulating hormone, Leydig cell number, the expression of Leydig and Sertoli cell genes and proteins were investigated. DNHP significantly increased serum testosterone levels at 10 mg/kg but lowered its level at 1000 mg/kg. DNHP significantly increased luteinizing hormone levels at 1000 mg/kg without affecting follicle-stimulating hormone levels. DNHP increased Leydig cell number at all doses but down-regulated the expression of Lhcgr, Hsd3b1, Hsd17b3, and Hsd11b1 in Leydig cell per se at 1000 mg/kg. DNHP elevated phosphorylation of ERK1/2 and GSK-3β at 10 mg/kg but decreased SIRT1 and PGC-1α levels at 1000 mg/kg. In conclusion, DNHP exposure causes Leydig cell hyperplasia possibly via stimulating phosphorylation of ERK1/2 and GSK-3β signaling pathways.