Mouse Testicles Research Articles

Expression and localization of Cyclin D1/Nanog and NF-κB/Bax protein in dysplastic testicles of mice

Testicular dysplasia significantly impairs male reproductive capacity. This study investigated the expression of Cyclin D1/Nanog and NF-κB/Bax in dysplastic testes of mice using histological staining, Western blotting, and immunohistochemistry. The results showed that Nanog and Bax expression were significantly higher in dysplastic testicular tissue than in normal tissue (P < 0.01). Cyclin D1 protein expression was higher in normal testis tissue than in dysplastic testis (P < 0.01). NF-κB was highly expressed in cryptorchid and normal testis with no significant difference (P > 0.05). Immunolocalization revealed that Nanog, NF-κB, and Bax were expressed in the cytoplasm of Leydig and spermatogenic cells. Cyclin D1 primarily expressed in the nucleus of Sertoli cells. These findings suggest that altered expression of Nanog, Cyclin D1, and Bax may contribute to testicular dysplasia. This study provides a scientific foundation for detecting testicular dysplasia and selecting appropriate animal models, ultimately informing strategies to improve male reproductive health.

The RNA-seq mapping of Testicular Development after Heat Stress in Sexually Mature Mice

The testis serves as the primary site for spermatogenesis in mammals and is a crucial organ for the secretion of male hormones. Heat stress (HS) can have adverse effects on the seminiferous tubules, sperm quality, and sperm fertilization capability within the testis. Despite numerous previous studies describing various time points after heat stress in mice, a systematic and comprehensive dataset on heat stress and recovery in mice has been lacking. This study aimed to explore the gene expression changes in the recovery of multiple seminiferous epithelial cycles and spermatogenic cycles in mouse testicles after heat stress. We obtained high-throughput bulk RNA-seq data from testicular tissue of 4 NC mice and 32 HS mice (divided into 9 groups: NC, 30 min, 2 h, 6 h, 24 h, 3d, 8d, 24d, 47d, and 95d) and illustrated the dynamic changes in differential genes. This data set provides valuable insights into the detailed dynamic changes of one or more spermatogenic cycles after heat stress in mouse testicles, as well as the molecular mechanisms involved.

Thymoquinone effects on autophagy, apoptosis, and oxidative stress in cisplatin-induced testicular damage in mice.

In this study, the effect of thymoquinone (TQ) on CP-induced spermatogenesis defects in mice has been investigated. Sperm parameters, serum testosterone concentration, histology, Bax/Bcl-2 ratio, and expression of autophagy-related biomarkers have been assessed. Total antioxidant capacity (TAC), total oxidant status (TOS), and oxidative stress index (OSI) in testicular tissue were examined for the evaluation of oxidative stress levels. CPhas induced histological changes and significantlyincreased the Bax/Bcl-2 ratio, decreased testosterone concentration, testicular weight, and sperm quality. CP induced oxidative stress by elevating OSIin the testicular tissue(p < 0.05).Expression of theautophagy-inducer genes (ATG7, ATG5, and Beclin-1) andratio ofLC3B/LC3Aproteinswere significantly decreased, while mTOR expression was increased in the CP group.TQ pretreatment dose-dependently decreased the Bax/Bcl-2 ratio and mTOR gene expression while increasing the expression of ATG5 and ATG7 genes, LC3B/LC3A ratio, and Beclin-1 proteins.TQ could also dose-dependently reverse the histology, testosterone level, and sperm quality of the CP-intoxicated mice. These findings show that TQ pretreatment can enhance sperm production by inducing autophagy and reducing apoptosis and oxidative stress in the CP-intoxicated mouse testicles.

Evaluation of the Potential Protective Role of Galangin Associated with Gold Nanoparticles in the Histological and Functional Structure of Testes of Adult Male Albino Mice Administrated with Carbon Tetrachloride

Galangin (Gal) is a natural flavonoid sourced from the roots of the plant Albina Kalanga. It possesses a variety of pharmacological activities, such as antioxidant, anticancer, and anti-inflammatory. The current study aims to enhance the efficacy of galangin through the use of gold nanoparticles as a drug delivery system against CCl4-induced toxicity in the testicular tissue of male albino mice. Forty-two albino mice were divided into seven groups (6 mice/group), treatment with carbon tetrachloride solution for two weeks by intraperitoneal injection, (1 ml/kg) once a week for all groups except the control group, after which a group was injected with gold nanoparticles and two groups were injected with galangin in two concentrations, and two groups were injected with AuNPs + Gal conjugation solution with two concentrations. The animals' bodies were weighed and blood samples were obtained for testosterone hormone analysis and testicles for the purpose of weighing and histological study at the end of the experiments. The study showed that the testicular tissue of mice treated with CCl4 had a different pathology compared to the control group, the testosterone hormone levels in the CCl4 group were significantly higher than those of the control group. Results from the AuNPs+Gal group showed a significant reduction in the effects of CCl4 toxicity on mice testicles, with testosterone levels are returning to normal and histological testicle structures improving, the ratio of testicle weights to animal body weight in the group injected with AuNPs was significantly higher than that of the control and CCl4 groups. In conclusion, the results revealed that galangin combined with gold nanoparticles could effectively reduced the histological and functional tissue damages caused by CCl4, presenting a promising natural solution that needs to be further developed

Ultrastructure of mice testes affected by orally administered crocin of crocus sativus

In this study, thirty-two adult male mice were divided into four groups control and experimental treatment groups received crocin at 4, 20, and 100 mg/kg daily for six weeks. In results, testosterone level was increased significantly from 18.5 ng/dl in the control to 49.3 ng/dl in group received crocin at 20 mg/kg. The regular features of seminiferous tubules with increased Sertoli cells have been observed in the 20mg/kg crocin group.&#x0D; In the group received crocin at 100 mg/kg, many of the seminiferous tubules were atrophic, spermatogenic cells were discontinuous in the same areas while the space among them has been increased, and fewer interstitial tissue between seminiferous tubules were observed. The changes were markedly advanced by increasing the number of atrophied seminiferous tubules.&#x0D; Electron microscopic results showed the large mitochondria with normal crista and dilated rough endoplasmic reticulum in spermatid cells of the 20 mg/kg crocin group.&#x0D; In the 100 mg/kg crocin group, most of the cell cytoplasm showed vacuolated mitochondria with patchy matrix and distorted vision cristae, with many lysosomes, lipid droplets and vacuoles. Some sperms had abnormal swollen nuclear and irregular wavy acrosomal system.&#x0D; The results of the present study showed that crocin at 20 mg/kg has a protective effect on the germinal epithelium of seminiferous tubules while at 100 mg/kg had destructive effects on the histological structure of the treated mice testicles.&#x0D; In conclusion, treating healthy male mice with crocin for six weeks increased the testosterone concentration while at 100 mg/kg damaged the testicle’s structure

The protective effect of Ozone on the mice testicular damage induced by methotrexate.

Methotrexate (MTX) is widely administered for the treatment of various cancers. However, MTX induces male reproductive toxicity. In the current study, the effect of ozone therapy (OT) on reducing the toxic effects of MTX in the mouse testicles has been investigated. Twenty-four mice were divided into four groups: control, OT (4 mg/kg ozone), MTX (20 mg/kg), and MTX + OT. Testosterone levels, histological changes, and oxidative stress biomarkers were assessed to evaluate the protective effects of OT. The results demonstrated that MTX disrupted germinal epithelium, reduced serum testosterone levels, and enhanced oxidative stress in testicular tissue. However, treatment with OT attenuated these adverse effects. OT effectively restored the levels of antioxidant enzymes, such as catalase (CAT), glutathione (GSH), and superoxide dismutase (SOD). OT reduced lipid peroxidation, as indicated by decreased malondialdehyde (MDA) levels. OT preserved normal spermatogenesis, improved morphometric parameters, and reduced histological changes by MTX. Moreover, OT effectively restored testosterone levels. OT protects against MTX-induced testicular damage by suppressing oxidative stress.

EphB4 and ephrin-B1 expression in the intra-testicular-resident macrophages in mice.

The objective was to find out the expression of EphB4 receptor and ephrin-B1 ligand by the macrophages that live inside the mouse testicles. Messenger ribonucleic acid (mRNA) expression of EphB4 and ephrin-B1 was identified via RT-PCR amplification, and protein expression was examined by immunostaining. Analysis using RT-PCR revealed that mRNA of EphB4 and ephrin-B1 were noticed in the examined testis of all postnatal ages. Furthermore, immunostaining revealed that F4/80-positive intra-testicular-resident macrophages were located in the intertubular spaces within the testis and more densely around the intra-testicular excurrent duct system, and increased in number gradually during the postnatal period of development until 5 weeks of age, when the mice attain their maturity (puberty), and maintained thereafter. Both EphB4 and ephrin-B1 immunoreactivity were noticed in F4/80-positive intra-testicular-resident macrophages within the testis of all studied postnatal ages. Ephrin-B1 and EphB4 immunoreactivity were weak during early postnatal development until the age of 2 weeks, and then ephrin-B1 immunoreactivity became very strong and EphB4 immunoreactivity became strong at the age of 3 weeks, and they continued to do so until the age of 8 weeks. Furthermore, EphB4 receptor was tyrosine-phosphorylated in testis. The expression of EphB4 and ephrin-B1 in mice intra-testicular-resident macrophages is being examined for the first time in this work. The localization of EphB4 and ephrin-B1, and EphB4 tyrosine-phosphorylation suggest that EphB4/ephrin-B1 signaling might occur in the intra-testicular-resident macrophages, and may participate in maintaining male fertility.

Marrubium alysson L. Ameliorated Methotrexate-Induced Testicular Damage in Mice through Regulation of Apoptosis and miRNA-29a Expression: LC-MS/MS Metabolic Profiling

Despite the efficient anti-cancer capabilities of methotrexate (MTX), it may induce myelosuppression, liver dysfunction and testicular toxicity. The purpose of this investigation was to determine whether Marrubium alysson L. (M. alysson L.) methanolic extract and its polyphenol fraction could protect mouse testicles from MTX-induced damage. We also investigated the protective effects of three selected pure flavonoid components of M. alysson L. extract. Mice were divided into seven groups (n = 8): (1) normal control, (2) MTX, (3) Methanolic extract + MTX, (4) Polyphenolic fraction + MTX, (5) Kaempferol + MTX, (6) Quercetin + MTX, and (7) Rutin + MTX. Pre-treatment of mice with the methanolic extract, the polyphenolic fraction of M. alysson L. and the selected pure compounds ameliorated the testicular histopathological damage and induced a significant increase in the serum testosterone level and testicular antioxidant enzymes along with a remarkable decline in the malondialdehyde (MDA) level versus MTX alone. Significant down-regulation of nuclear factor kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), p53 and miRNA-29a testicular expression was also observed in all the protected groups. Notably, the polyphenolic fraction of M. alysson L. displayed a more pronounced decline in the testicular levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and MDA, with higher testosterone levels relative to the methanolic extract. Further improvements in the Johnsen score, histopathological results and all biochemical assays were achieved by pre-treatment with the three selected pure compounds kaempferol, quercetin and rutin. In conclusion, M. alysson L. could protect against MTX-induced testicular injury by its antioxidant, anti-inflammatory, antiapoptotic activities and through the regulation of the miRNA-29a testicular expression. The present study also included chemical profiling of M. alysson L. extract, which was accomplished by LC-ESI-TOF-MS/MS analysis. Forty compounds were provisionally assigned, comprising twenty compounds discovered in the positive mode and seventeen detected in the negative mode.

Biochemical and histopathological investigation of the protective effects of melatonin and vitamin E against the damage caused by acetamiprid in Balb-c mouse testicles at light and electron microscopic level.

The protective effects of melatonin (Mel) and vitamin E (Vit E) against the negative effects of acetamiprid (Acmp) on testicles, reproductive hormones, and oxidative stress parameters were investigated in the present study. A total of 50 Balb-c male mice were used in 7 groups; 6 mice in the control groups (distilled water, corn oil, ethanol), and 8 in other groups (Acmp, Acmp + Mel, Acmp + Vit E, Acmp + Vit E + Mel). After the experiment, which lasted 21days, hematoxylin eosin (H&E), periodic acid Schiff (PAS), and caspase-3 immunohistochemical (IHC) staining was performed on the testicular tissues. Also, the tissues were examined ultrastructurally with the transmission electron microscopy (TEM). In the Acmp group, there were decreased seminiferous tubule diameter and epithelial thickness, epithelial degeneration, decreased spermatozoa in the lumen, decreased PAS-positive staining in the seminiferous epithelial basement membrane, edema in the interstitial area, and hydropic degeneration in Leydig cells. Caspase-3 immunoreactivity was higher than in the other groups. TEM examination showed degeneration in tubule cells, lysosomal accumulation in cells of the spermatogenic line, vacuolizations with myelin figures, and necrosis. Hydropic degeneration, electron-dense lipid vacuoles, and chromatolysis were evident in the Leydig cell cytoplasm. In Sertoli cells, electron-dense lysosomal deposits were noted. In biochemical terms, there were decreased tissue glutathione (GSH) and total antioxidant status (TAS), and increased malondialdehyde (MDA) and total oxidant status (TOS). Plasma luteinizing hormone (LH), follicular stimulating hormone (FSH), and testosterone levels were decreased. In the groups with melatonin, vitamin E, and both were applied together, tissue damage, and apoptotic cell death were reduced at both light microscopic and ultrastructural levels. In biochemical terms, there were decreased oxidative parameters and increased hormonal parameters. It was found that vitamin E was more effective in decreasing oxidative parameters and increasing antioxidative parameters when compared to melatonin, and hormonal parameters increased at a higher level in the Acmp + Vit E group than in all groups. As a result, it was found that exposure to Acmp caused damage to testicular tissue, induced oxidative stress in testicles, and decreased plasma LH, FSH, and testosterone levels, and although vitamin E is more effective than melatonin in preventing this damage, both are effective.

Highly sensitive in vitro bioassay for luteinizing hormone and chorionic gonadotropin allowing their measurement in plasma.

In previous studies, we had shown the synergistic effect of 10−5 M forskolin (FSK) on the detection threshold of the cyclic AMP response to luteinizing hormones (LH) and chorionic gonadotropins (CG) from various species in the mouse Leydig tumor cell (mLTC) cell line. Independently, we started to study the effect of 10−12–10−6 M oxytocin (OXT) also on the cyclic AMP response to LH and CG preparations on these same cells and found an amplifying effect on the luminescence response caused by gonadotropins. The aim was then to explore the effects of 10−12–10−6 M OXT on the gonadotropin-induced cAMP response, in the presence or absence of 10 µM FSK to optimize the assay down to a sensitivity compatible with the detection of the circulating concentrations of these hormones in various species. Finally, the optimization relies on three independent phenomena: (1) the inhibition of nucleotide phosphodiesterase by IBMX (3-isobutyl-1-methylxanthine) to avoid cAMP degradation; (2) the strong synergy of 10 µM forskolin with low concentrations of LH or CG during the 1-h luminescence measurement; (3) the stimulatory effect of 10−8M OXT on the amplitude of transfected cAMP-sensitive luciferase response. By doing this, the detectable concentrations are at the 1–10 pg/well (pM range) for the LHs and CGs from various species. The bioactivities of circulating LHs and CGs in blood or urine are therefore expected to be measurable in 10 µL-plasma samples from mammalian species and maybe others. Indeed, a preliminary study with equine and donkey plasma samples shows that the measured bioactivity was fully inhibited by a specific MAB against the receptor-binding region of equine LH (eLH) and equine CG (eCG), thus eliminating a possible response due to interfering substances other than eLH or eCG. From these data, it is expected that the bioactivity profiles of these hormones will be measurable in the blood of human, equine, and ovine species and very likely in rodents, ruminants, and hopefully in most other mammalian species.Lay summaryLuteinizing hormone (LH) plays a central role in controlling ovary and testicle functions in many animals, including humans. The highly sensitive method, known as an assay, described in this paper, measures the biological activity of LH in the blood of mammals. The assay is performed in culture of cells derived from mouse testicles in the presence of factors that diminish the detection threshold for LH. The knowledge of the bioactive LH concentration dynamics in the blood is very informative about the reproductive status of male and female mammals. This new in vitro bioassay provides a powerful tool to get this information.

Interleukin-6 deficiency modulates testicular function by increasing the expression of suppressor of cytokine signaling 3 (SOCS3) in mice

Several cytokines have been reported to participate in spermatogenesis, including interleukin-6 (IL6). However, not many studies have been conducted on the loss of Il6 on the male reproductive tract. Nonetheless, there is considerable knowledge regarding the pathological and physiological role of IL6 on spermatogenesis. In this way, this study evaluated the impact of Il6 deficiency on mice testicles in the absence of infection or inflammation. We showed that Il6 deficiency increases daily sperm production, the number of spermatids, and the testicular testosterone and dihydrotestosterone levels. Besides that, mice with a deleted Il6 (IL6KO) showed increased testicular SOCS3 levels, with no changes in pJAK/JAK and pSTAT3/STAT3 ratios. It is worth noting that the aforementioned pathway is not the only pathway to up-regulate SOCS3, nor is it the only SOCS3 target, thus proposing that the increase of SOCS3 in the testis occurs independently of the JAK-STAT signaling in IL6KO mice. Therefore, we suggest that the lack of Il6 drives androgenic production by increasing SOCS3 in the testis, thus leading to an increase in spermatogenesis.

Effect of a Freezing Medium Containing Melatonin on Markers of Pre-meiotic and Post-meiotic Spermatogonial Stem Cells (SSCs) After Transplantation in an Azoospermia Mouse Model Due to Testicular Torsion.

Spermatogonial stem cells (SSCs) are essential to the initiation of spermatogenesis. Cryopreservation, long-term maintenance, and auto-transplantation of SSCs could be a new treatment for infertility. The aim of this study was to add melatonin to the basic freezing medium and to evaluate its effect on the efficiency of the thawed SSCs after transplantation into the testicles of azoospermic mice. SSCs were isolated from newborn NMRI mice, and the cells were enriched to assess morphological features. The thawed SSCs were evaluated for survival, apoptosis, and ROS level before transplantation, and the proliferation (MVH and ID4) and differentiation (c-Kit, SCP3, TP1, TP2, and Prm1) markers of SSCs were examined using immunofluorescence, western blot, and quantitative real-time polymerase chain reaction (PCR) after transplantation. It was found that the survival rate of SSCs after thawing was significantly higher in the melatonin group compared with the cryopreservation group containing basic freezing medium, and the rate of apoptosis and level of ROS production also decreased significantly in the cryopreservation group with melatonin (p < 0.05). The expression of proliferation and differentiation markers after transplantation was significantly higher in the cryopreservation group with melatonin compared to the cryopreservation group (p< 0.05). The results suggest that adding melatonin to the basic freezing medium can effectively protect the SSCs by increasing the viability and reducing the ROS production and apoptosis and improve the transplantation efficiency of SSCs after cryopreservation, which will provide a significant suggestion for fertility protection in the clinic.

The Effect of Coffee Fruit Skin Extract on Sperm Characteristics And Testicular of Mice With Ethanol-Induced

Coffee fruit skin contains antioxidant compounds that can repair damaged tissues, especially those of reproductive or-gans. This study was designed to assess to what extent the metha-nolic extract of coffee fruit skin affected sperm characteristics and testicular histology in mice receiving 15% ethanol for 15 days. It used 25 male mice aged four months and employed a completely randomized design with two controls (K) and three treatments (P), namely K1 (without ethanol), K2 (15% ethanol), P1 (15% ethanol and the methanolic extract of coffee fruit skin, or MECS, at 125 mg/ kg BW), P2 (15% ethanol and MECS 250 mg/kg BW), and P3 (15% ethanol and MECS 500 mg/kg BW). These treatments were administered orally with a 1 mL disposable syringe for 15 days. The parameters observed were sperm characteristics (viability, morphology, and sperm count) and the microscopic structure of the testicles (lu-men area, diameter, and area of seminiferous tubules). These data were then analyzed using the one-way ANOVA, continued with the LSD and Duncan’s tests (P &lt;0.05). The results showed that P2 (250 mg/kg BW) substantially improved sperm count (86.5x105±1.73 cells/ mL), sperm viability (87.26±0.05%), and the proportion of normal sperm morphology (93.33±0.004%) of mice with 15% ethanol-in-duced sperm damages. Testicular histology also confirmed improve-ments in spermatogenic cells, as evident from the lumen, diameter, and area seminiferous tubules after receiving P1 and P2. In conclu-sion, the optimum dose of the coffee skin extract for improving sperm quality and microscopic structures of mice testicles is 250 mg/kg BW.

Evaluación estacional de la reproducción de esteroides sexuales en testiculos del ratón de orejas oscuras (Peromyscus melanotis Allen &amp; Chapman, 1897) de diferentes clases de edad

Se realizó un estudio sobre la distribución local, altitudinal y por comunidad vegetacional de las mariposas de la superfamilia Papilionoidea en 37 localidades de las cuatro entidades que comprende la Sierra Nevada. El área se encuentra entre los 1900 y los 3800 msnm y presenta las siguientes comunidades vegetacionales: bosque mesófilo de montaña, de oyamel, Alnus, encino, encino-pino, pino, pino-encino, bosque caducifolio, selva baja caducifolia-bosque mesófilo de montaña, vegetación perturbada y cultivos. En este trabajo se registraron 75especies, de las cuales 34 son nuevos registros para el área. Se integró una relación actualizada de 85 especies con base en la literatura y en los registros obtenidos en el trabajo de campo. El mayor número de especies correspondió a la familia Nymphalidae. En el análisis espacial de los papilionoideos se encontró que la mayor riqueza y abundancia ocurrió en la localidad de Santa Catarina, en el Estado de México. Se observó que aunque la riqueza específica entre la vertiente occidental y la oriental fue la misma (58 especies), su composición faunística es distinta. Con respecto a la distribución por vegetación, se encontró que en la vertiente oriental las especies de Papilionoidea mostraron los valores más altos y que el bosque mesófilo de montaña y el de encino fueron las comunidades más ricas y abundantes de las 11 reconocidas en el estudio.

Urine-Derived Stem Cells Facilitate Endogenous Spermatogenesis Restoration of Busulfan-Induced Nonobstructive Azoospermic Mice by Paracrine Exosomes.

Nonobstructive azoospermia (NOA) is a severe form of male infertility, with limited effective treatments. Urine-derived stem cells (USCs) possess multipotent differentiation capacity and paracrine effects, and participate in tissue repair and regeneration. The aim of this study is to investigate whether the transplantation of USCs or USC exosomes (USC-exos) could promote endogenous spermatogenesis restoration in a busulfan-induced NOA mice model. USCs were cultured and characterized by flow cytometry. High-density USCs were cultured in a hollow fiber bioreactor for exosomes collection. USC-exos were isolated from USCs conditional media and identified by transmission electron microscopy, western blotting, and Flow NanoAnalyzer analysis. USC-exos exhibited sphere- or cup-shaped morphology with a mean diameter of 66.5 ± 16.0 nm, and expressed CD63 and CD9. USCs and USC-exos were transplanted into the interstitial space in the testes of NOA mice per the following groups: normal group; groups treated with no injection, phosphate-buffered saline (PBS), USCs or USC-exos on days 3 and 36 after busulfan administration, respectively. Thirty days after USCs and USC-exos transplantation, spermatogenesis was restored by both USCs and USC-exos in NOA mice 36 days after busulfan treatment as confirmed by immunofluorescence staining and hematoxylin and eosin staining. Moreover, spermatogenic genes (Pou5f1, Prm1, SYCP3, and DAZL) and the spermatogenic protein UCHL1 were significantly increased in both the USCs 36 and USC-exos36 groups compared with the PBS group, as demonstrated using quantitative real-time polymerase chain reaction and western blot analysis. However, the transplantation of USCs or USC-exos at day 3 after busulfan treatment did not improve spermatogenesis in NOA mice. Our study demonstrated that USCs could facilitate endogenous spermatogenesis restoration of busulfan-induced NOA mice through paracrine exosomes but could not protect the mouse testicles at the early stage of destruction caused by busulfan. This study provides a novel insight into the treatment of NOA.

Protective effect of Zingerone against mouse testicular damage induced by zinc oxide nanoparticles.

The purpose of the present study was to evaluate the effect of Zingerone (Zing) on zinc oxide nanoparticle (ZNP)-induced spermatogenesis defects in mice. To this end, 50mg/kg of ZNP was prescribed to the mice as an intoxicated group for 35days. In protection groups, Zing (10, 20, and 40mg/kg) was given prior to ZNP treatment for seven days and then co-administration of ZNP for 35days. Epididymal sperm parameters, testicular histology, Johnsen's scoring, morphometric parameters, TUNEL staining, oxidative stress, and serum testosterone level were evaluated for determining ZNP and Zing effects on the mouse testicles. Effects of Zing and ZNP on the viability of mouse Leydig (TM3) and mouse Sertoli (TM4) cell lines were also done. Testicular weights, testosterone levels, sperm quality, morphometric parameters, Johnsen's score, and superoxide dismutase (SOD) and catalase (CAT) activities were significantly decreased in ZNP-intoxicated mice, while apoptotic index, Malondialdehyde (MDA) content, and histological features, including epithelial vacuolization, sloughing, and germ cell detachment, were improved significantly in ZNP-intoxicated mice. Pretreatment with 20 or 40mg/kg Zing significantly reduced the histological criteria, increased morphometric parameters, enhanced testosterone levels, attenuated apoptotic index, improved sperm quality, and reversed oxidative stress by reducing the level of MDA and incrementing the activity level of SOD and CAT enzymes. Zing dose-dependently enhanced the viability of ZNP-treated TM3 and TM4 cells in comparison with only ZNP-exposed cells. According to the results of our study, Zing effectively prevented the defects in spermatogenesis among mice treated by ZNP.

Growing Trans-Species Islets in Tumor Extract-Remodeled Testicles.

Although pancreatic islet transplantation holds promise for the treatment of type I diabetes, its application has been significantly hampered by transplant rejection. Here, an approach is demonstrated to support trans‐species islet beta cells from a rat to grow and function in the body of a mouse host while overcoming graft rejection. This approach, which builds on remodeling of the mouse testicle by local injection of a tumor homogenate, establishes an immunosuppressive and proregenerative niche in the testicle. This remodeling proves necessary and effective in shaping the testicle into a unique site to accommodate xenograft cells. Rat pancreatic beta cells—from both the insulinoma (cancer cells) and pancreatic islet (normal tissue)—survive, grow, and form a desirable morphology in the remodeled mouse testicle. Notably, when hyperglycemia is induced in the host body, these xenografts secrete insulin to regulate the blood glucose level in mice for as long as 72 days. Furthermore, no graft rejection, acute inflammation, or safety risks are observed throughout the study. In summary, it is demonstrated that the growth of xenogeneic insulinoma cells in a mouse testicle might serve as an alternative approach for islet transplantation.

A novel role for zinc transporter 8 in the facilitation of zinc accumulation and regulation of testosterone synthesis in Leydig cells of human and mouse testicles

ObjectiveZinc is intimately involved in testosterone production. Zinc transporter 8 (ZnT8) is found to be localized in insulin secretory granules as a β-cell specific Zn transporter. The effect of ZnT8 and related zinc accumulation in steroidogenesis, however, is still unknown. The present study aimed to explore whether ZnT8 plays a role in the facilitation of zinc accumulation and regulation of testosterone synthesis in testicles. MethodsLeydig cells were isolated from the testicles of human, CD-1 suckling and ZnT8-KO mice. Zn accumulation in mitochondria was induced by hCG stimulation. Transfection of hZnT8-EGFP and RNA interfere of mZnT8 were done in MLTC-1 cells. ZnT8 expression and its co-localization with steroidogenic acute regulatory (StAR) protein were analyzed with RT-PCR, Western blot and dual-fluorescent staining protocols. Serum testosterone levels in mice were determined with chemiluminescent enzyme immunoassay. ResultsZnT8 was found to be presented in Leydig cells and up-regulated in suckling mouse Leydig cells and MLTC-1 cells after hCG administration, by which zinc accumulation occurred in mitochondria. ZnT8 gene silencing or knockout inhibited stimulated progesterone and testosterone production, reduced stimulated zinc accumulation and down-regulated phosphorylated steroidogenic acute regulatory (StAR) expression in Leydig cells. Furthermore, an inhibitor (H89) of PKA blocked hCG-stimulated progesterone caused by ZnT8 over-expression and zinc treatment. ConclusionThe present study provided the first evidence that ZnT8 transports Zn into Leydig cell mitochondria with gonadotropin stimulation and suggests that ZnT8 may play a role in testosterone production via the PKA signaling pathway.

A Homeopathic Specialty Protects from the Mutagenic Effects of Mitomycin C

A homeopathic specialty (DIG) composed of three herbal stocks, Berberis vulgaris, Taraxacum officinale and Arctium lappa at different dilution levels, was assessed for its antimutagenic properties against mitomycin C. The micronucleus assay on Chinese hamster ovary (CHO)-K1 cells was used to evaluate the in vitro anticlastogenic activity of DIG compared with those of separately diluted mother tinctures. The micronucleus assay was performed on mouse erythrocytes, and the comet assay was performed on mouse liver, kidney, lung, brain, and testicles to assess the protective effects of DIG (0.2 and 2% at libitum) against an intraperitoneal injection of mitomycin C (1 mg kg-1) in mice. DIG exerted a powerful anticlastogenic activity under both pretreatment and simultaneous treatment conditions as assessed by the micronucleus assay in CHO-K1 cells with protective ratios reaching 74.1 and 55.5%, respectively. Its protective activity was greater than that observed for each mother tincture. DIG reduced micronuclei levels in mouse erythrocytes and suppressed 80% of DNA strand breaks in the liver, kidney, lung, brain, and testicles of mice exposed to mitomycin C. In conclusion, this specialty showed a great protective activity both in vitro and in vivo against the mutagenic effects of mitomycin C. These first results are promising regarding the follow-up of patients under chemotherapy.

Evaluate the Influence of Eupatorium adenophorum Extract with Mice Organ

In order to study the influence of extract from Eupatorium adenophorum in mice organs, this experiment will be the basis of further study that make Eupatorium adenophorum become Phyto contraceptive, this experiment take the feeding respectively way after the completion of the 1D, 5D, 10d, 15d of Eupatorium adenophorum mice by intragastrical administration of levonorgestrel group and blank control group. After the same operation in different periods of small rat heart and kidney the uterus, testis, and other organs were observed. The results showed that after extraction of E. adenophorum changes in female mice uterus shape was perfused significantly, showed swelling larger. Data analysis of each viscera coefficient was found E. adenophorum had No obvious effect on the heart, kidneys and testicles of mice. but there are obvious differences date between the treatment group and the blank group. (5d: F=10. 800 P=0. 043 cases) from tissue sections we can see female mice uterus cell morphology changes significantly, there was a similar appearance change in the uterus of the female mice with the estradiol For a male mouse testis of E.adenophorum gavage had No obvious effect. And it is found that the heart, the treated mice kidney, testis, ovary and other organs were observed in each period of time the organization had No obvious change; only female mice uterus tissue sections of individual cells became larger, and the organization of the gap larger. This research shows that E.adenophorum extract has the potential to develop botanical contraceptives, we will conduct in-depth study.