Cells In Testicular Tissue Research Articles

Conserved Sex-Specific Timing of Meiotic Initiation During Sex Differentiation in the Protandrous Black Porgy Acanthopagrus schlegelii

Meiosis is an essential mechanism of gametogenesis for all sexually reproducing species. In vertebrates, one conserved aspect of sex differentiation is that female embryonic germ cells enter meiosis earlier than male germ cells. In some lower vertebrates, female germ cells proliferate prior to entering meiosis, whereas male cells remain in mitotic arrest. Protandrous black porgy fish, Acanthopagrus schlegelii, have a dramatic life cycle involving a characteristic sex change. Black porgy are functional males for their first and second spawning seasons, but approximately half of the fish transform into females during their third year. We cloned the black porgy homologs of dosage suppressor of mck1 homolog (dmc1) and synaptonemal complex protein 3 (sycp3), and examined their expression profiles as well as those of cytochrome P450 family 26 genes (cyp26: cyp26a and cyp26b), retinaldehyde dehydrogenases (raldh: raldh2 and raldh3), retinoic acid receptors (rars: raralpha, rarbeta, rargamma, and rargammab), retinoid X receptors (rxrs: rxralpha, rxrbeta, and rxrgamma) and deleted azoospermia-like (dazl) during gonadal sex differentiation by RT-PCR, quantitative RT-PCR, and immunohistochemistry. Our results show that during gonadal development, germ cells located in ovarian tissue proceed into meiosis earlier than germ cells in testicular tissue. Furthermore, treatment with estradiol-17beta, which induced cyp26 expression, blocked dazl and raldh expression and reduced the expression of rars, rxrs, dmc1, and sycp3. This unique model therefore suggests that the temporal differences in meiosis initiation between females and males are conserved during gonadal sex differentiation in hermaphroditic vertebrates.

The effects of honey and vitamin E administration on apoptosis in testes of rat exposed to noise stress

AIMS:A variety of stress factors are known to inhibit male reproductive functions. So this study was conducted in order to investigate the effects of honey and vitamin E on the germinative and somatic cells of testes of rats exposed to noise stress.MATERIALS AND METHODS:Mature male wistar rats (n = 24) were randomly grouped as follows: Group 1 (honey + noise stress), 2 (vitamin E + noise stress), 3 (noise stress,) and 4 as the control group. In groups 1, 2, and 3, rats were exposed to noise stress. In groups 1 and 2, rats also were given honey and vitamin E, respectively, orally for 50 days. After that, the germinative and somatic cells of testes parenchyma were isolated by digesting the whole testes by a standard method. Next, viability, apoptosis, and necrosis of the cells were evaluated by TUNEL kit and flow cytometry.RESULTS:The rates of apoptosis and necrosis of the testicular cells were increased (P = 0.003 and P = 0.001, respectively), but viability of these cells decreased in testes of rats exposed to noise stress (P = 0.003). However, administration of honey and vitamin E were significantly helpful in keeping the cells of testis parenchyma alive, which suffers from noise pollution (P < 0.05 and P < 0.05, respectively).CONCLUSIONS:Noise stress has negative influences on the cells of testicular tissue by increasing apoptotic and necrotic cells. However, the associated enhancement in healthy cells suggests that honey and vitamin E have positive influences on the testis parenchyma.

Screening for partial AZFa microdeletions in the Y chromosome of infertile men: is it of clinical relevance?

To evaluate the frequency of complete and partial AZFa Y-chromosome microdeletions among infertile Israeli men. To review the published frequencies and histologic findings of AZFa deletions. Retrospective study. Academic medical center. A total of 1,260 infertile Israeli men. Literature review (2000-2010) of reports on men with AZFa deletions and their testicular findings. The DNA of 1,260 infertile men was evaluated for AZF microdeletions. The DNA of 657 of them with undetected microdeletions was analyzed for partial AZFa deletion in the USP9Y and DDX3Y genes using sequence-tagged sites beyond EAA/EMQN recommendations. The frequency of complete and partial AZFa microdeletions. Availability of sperm cells for intracytoplasmic sperm injection in men with complete/partial microdeletions. Two men had complete AZFa deletion (a frequency of 0.28% among nonobstructive azoospermic men). None had partial AZFa deletions. The likelihood of finding sperm cells in men with complete AZFa deletions is negligible. Complete AZFa deletion is rare and usually associated with azoospermia and absence of sperm cells in testicular tissue. The low frequency of partial AZFa deletions and the inconsistent prospects for spermatogenesis reported in the literature question the need for routine assessment of microdeletions in genes, such as USP9Y or DDX3Y.

18F-ML-10, a PET Tracer for Apoptosis: First Human Study

Clinical PET of apoptosis may have substantial value in advancing patient care. We report here the first-in-humans study with (18)F-labeled 2-(5-fluoropentyl)-2-methyl malonic acid ((18)F-ML-10), a small-molecule PET tracer for apoptosis. Presented are the dosimetry, biodistribution, stability, and safety profiles of this PET tracer in healthy human volunteers. Also reported is tracer binding to targeted apoptotic cells in testicular tissue, where a relative abundance of apoptotic cells is normally observed. (18)F-ML-10 (233 ± 90 MBq) was intravenously administered to 8 healthy subjects, followed by whole-body PET/CT for 220 min. Serial blood and urine samples were collected for radioactivity measurement, and plasma tracer stability was assessed by high-performance liquid chromatography. Dosimetry calculations were performed using OLINDA/EXM software. (18)F-ML-10 manifested high stability in vivo and rapid distribution followed by fast clearance, with an elimination half-life of 1.3 ± 0.1 and 1.1 ± 0.2 h from the blood and from all other organs, respectively, and excretion through the urine. Dosimetry showed an average effective whole-body dose of 15.4 ± 3.7 μSv/MBq, with the urinary bladder being the dose-limiting organ. Selective accumulation and retention of the tracer in the testes was observed in all male subjects, a finding also demonstrated in mice using both small-animal PET and histopathology, confirming binding to apoptotic cells. Administration of (18)F-ML-10 was safe, without adverse effects. (18)F-ML-10 administered to healthy humans demonstrated a favorable dosimetry, biodistribution, stability, and safety profile. Binding to apoptotic sites was also demonstrated. These data support further development of this small-molecule probe for clinical PET of apoptosis.

CDY1 and BOULE transcripts assessed in the same biopsy as predictive markers for successful testicular sperm retrieval

To evaluate the molecular markers CDY1 and BOULE in the same testicular biopsy for predicting success of sperm retrieval in azoospermic men. Prospective study. University-affiliated medical center. Azoospermic men (n = 92) who underwent testicular sperm extraction (TESE) and who were classified as normal spermatogenesis, mixed atrophy, spermatocyte maturation arrest, or Sertoli cell only according to their combined histological and cytological testicular findings. Quantitative and qualitative evaluation of testicular biopsies by histological and qualitative reverse transcriptase-polymerase chain reaction (RT-PCR) expression methodologies. CDY1 and BOULE expression and the presence of sperm cells in testicular tissue. Both transcripts significantly predicted the presence of sperm cells by qualitative and quantitative methodologies. Although CDY1 had the best sensitivity by qualitative RT-PCR (98.3%), assessing both transcripts simultaneously had an additive efficacy compared with assessing CDY1 alone, improving the specificity from 84.4% to 96.3%. Assessing the expression of both CDY1 and BOULE by qualitative RT-PCR is a sensitive and feasible test for predicting the presence of sperm cells in testicular tissue and may serve as a predictive tool if repeated TESE is required.

The effects of molsidomine on hypoxia inducible factor alpha and Sonic hedgehog in testicular ischemia/reperfusion injury in rats

This study was designed to determine the effect of molsidomine (MO), a precursor of nitric oxide (NO) donor, on hypoxia inducible factor alpha (HIF-1alpha) and Sonic hedgehog (Shh) levels considered to be involved in the development of testes ischemia/reperfusion (I-R) injury. Torsions were created by rotating ipsilateral testes 720 degrees in a clockwise direction for 6 h and 1-h detorsion of the testis was performed. A sham operation was performed in group 1 (control, n = 7). In group 2 (I-R/Untreated, n = 7), following 6 h of unilateral testicular torsion, 1-h detorsion of the testis was performed. No drug was given. In group 3 (I-R/MO), after performing the same surgical procedure as in group 2, a NO donor MO was given at the starting time of reperfusion. In group 4 (I-R/L-NAME), after performing the same surgical procedure as in group 2, L-NAME was given at the starting time of reperfusion. Testes malondialdehyde (MDA) levels were determined as well as examining the testes histologically. Treatment of rats with MO produced a significant reduction in the levels of MDA and histopathological score compared to testes I-R groups. The Sonic hedgehog (Shh) expression in the basement membrane of the tubuli seminiferi, and sertoli and germinal cells in testicular tissue, were greatly increased in the I-R/MO group compared to groups 1, 2 and 4. Additionally, the HIF-1alpha expression in the interstitial spaces in testicular tissue were greatly increased in the I-R/MO group. The results suggest that MO has a protective effect against ischemia/reperfusion injury in rat testes and may affect Shh and HIF-1alpha signaling pathway.

Lycopene protects against cyclosporine A-induced testicular toxicity in rats

Cyclosporine A (CsA)-induced direct failures in hypothalamic–pituitary–gonadal axis and Sertoli cell phagocytic function have been considered for testicular toxicity so far. It has clearly been reported that oxidative stress leads to damage in sperm functions and structure of the testis. Therefore, this study was conducted to demonstrate whether CsA causes testicular and spermatozoal toxicity associated with the oxidative stress, and to investigate the possible protective effect of lycopene against CsA-induced damages in all reproductive organs and sperm characteristics in male rats. While the daily administration of CsA at the dose 15 mg/kg for 21 days significantly decreased the seminal vesicles weight, epididymal sperm concentration, motility, testicular tissue glutathione (GSH), glutathione peroxidase (GSH-Px) and catalase (CAT), diameter of seminiferous tubules and germinal cell thickness, it increased malondialdehyde (MDA) level and abnormal sperm rates along with degeneration, necrosis, desquamative germ cells in testicular tissue. However, the CsA along with simultaneous administration of lycopene at the dose of 10 mg/kg markedly ameliorated the CsA-induced all the negative changes observed in the testicular tissue, sperm parameters and oxidant/antioxidant balance. In conclusion, CsA-induced oxidative stress leads to the structural and functional damages in the testicular tissue and sperm quality of rats and, lycopene has a potential protective effect on these damages.

Effect of acute hypobaric hypoxia on spermatogenesis and lactate concentration in testicular tissue of male albino rats

Parameters of spermatogenesis in male albino rats was studied after experimental acute hypobaric hypoxia. The number of spermatogenic epithelial cells, Sertoli cells, and Leydig cells in testicular tissue significantly decreased in the posthypoxic period. Single exposure to hypoxia led to significant changes in lactate concentration in the testicular tissue. Cell composition of the testicles did not return to normal by the 60th day after treatment.

The effects of nitric oxide on the expression of cell adhesion molecules (ICAM-1, UEA-1, and tenascin) in rats with unilateral testicular torsion

Background/Purpose: The aim of this study was to determine the effects of nitric oxide (NO) on the expression of adhesion molecules in the early course of testicular I-R injury in rats. Methods: Forty male Sprague-Dawley rats were separated into 4 groups, each containing 10 rats. A sham operation was performed in group 1 (control). In group 2 (I-R), after 6 hours of unilateral testicular torsion, 1-hour detorsion of the testis was performed. In group 3 (I-R/ l-NAME), after performing the same surgical procedures as in group II, l-NAME was given for 30 minutes. In group 4 (I-R/Mol), after performing the same surgical procedure (torsion and detorsion) as in group II, molsidomine, an NO donor, was infused for 30 minutes. Then, ipsilateral orchiectomies were performed to measure the tissue levels of malondialdehyde (MDA) and NO and to make histologic examination. Results: MDA values and the testicular injury score decreased and NO values increased in the I-R/Mol-treated group compared with other experimental groups. The tenascin expression in the interstitial space and basement membrane of the tubuli seminiferi were milder in the I-R/Mol group compared with that of the I-R and the I-R/ l-NAME. The acrosomes of the spermatids in I-R and I-R/ l-NAME groups were stained mildly by lectin. In the I-R and I-R/ l-NAME groups, the interstitial spaces, basement membrane of the tubuli seminiferi, and sertoli and germinal cells in testicular tissue were stained intensely by ICAM-1. Conclusions: The expression of adhesion molecules such as tenascin, lectin, and ICAM-1 in the torted testicular tissue may be a pathophysiologic sign of inflammation. NO regulates adhesion molecules expression.

Aging of the male reproductive system.

Reproductive and sexual physiology, changes in body composition and mental performance in the aging male cannot simply be reduced to presumptive hypogonadism defined by low androgen serum levels or by decreasing levels of growth hormone (GH) and melatonin. Morphological changes in organs at different regulatory levels of hormonal networks governing, for example reproduction, such as diminished hypothalamic pulse generator mass, focal degeneration and loss of Leydig cells in testicular tissue, lead to diminished reserve capacities in production and to loss of coordinated pulsatile release of hypothalamic neuropeptides (e.g. gonadotropin releasing hormone, GnRH) and consequently diminished release of pituitary protein and glycoprotein hormones and testicular steroid hormones. Owing to presumptive alterations in feedback sensitivity, decreased testosterone levels do not necessarily upregulate pituitary LH secretion. Alternatively, increased serum levels of LH and FSH can be observed in old men either because of primary hypogonadism or to decreased hypothalamic opioid tone. In general, endocrine functions are sufficient to maintain fertility in elderly men because, except for sperm motility, quantitative and qualitative functional semen parameters are apparently not affected by age. Nevertheless, reduced endocrine and organic functions might become critical at different levels, with high inter-individual variability, of the hypothalamo/pituitary/gonadal-axis. One of the most intriguing organic manifestations of male aging is benign prostatic hyperplasia (BPH), the pathologic prevalence of which closely matches age. Age-associated changes in the endocrine system and in local networks of epithelial, stromal and luminal factors may play important roles in BPH development.

Rapid method to detect CIS-cells.

Seminoma cells, like intratubular tumor cells (carcinoma in situ), are characterized by abundant cytoplasmic glycogen. Visualization of glycogen in histologic sections has been shown to be a valuable diagnostic tool for detecting tumor cells in testicular tissue provided that appropriate fixatives were used to preserve the glycogen in the cells. Tumor cells fixed with 5.5 % glutaraldehyde show virtually no loss of their highly soluble content. In combination with other structural characteristics even single tumor cells can be histologically diagnosed (Holstein et al., 1987).

Immuno-electron microscopic localization of calmodulin and calmodulin-binding proteins in the mouse germ cells during spermatogenesis and maturation

When extracts of mouse testis were Western-blotted against a monoclonal antibody which reacts with calmodulin in the presence of Ca2+, all calmodulin was associated with the macromolecules of molecular weight above 50 kDa. Immuno-electron microscopy of testes using this antibody indicated that calmodulin is localized at higher density in the nucleus and cytoplasm of germ cells during the developmental phase between pachytene and round spermatid, showing the highest level just before meiotic divisions. There was no special association of calmodulin to any organelles in these cells. Extremely low levels of calmodulin occurred in spermatogonia and other testicular tissue cells. Calmodulin decreased dramatically as spermatids underwent metamorphosis, becoming detectable only at the perinuclear space of sperm heads. Further relocation to the postacrosomal region occurred during sperm transit to the cauda epididymis. Immunodetection after the calmodulin overlay on ultrathin sections revealed a sharp increase of calmodulin immunogold deposits in the nuclei of spermatids accompanying their condensation. The results indicate that some calmodulin-binding proteins, but not calmodulin itself, accumulate in the nuclei during the final steps of spermiogenesis.