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PubMed2026

Post-translational modification of proteins in the human testis development pathway.

BACKGROUND: The foetal testes produce the androgens necessary to masculinise the developing embryo and support the maturation of germ cells, that will eventually develop into sperm, thus ensuring future reproductive capacity. The testes develop from the bi-potential gonads in a highly orchestrated process resulting in the differentiation of a complex tissue with multiple cellular lineages. While recent transcriptomic and chromatin-based analyses of human foetal testes have provided an unprecedented level of insight into signalling pathways activated during this process, proteomic studies of the human foetal gonads remain limited. Proteins are active molecules and post-translational modification (PTM) of proteins influences protein activity, stability and localisation. Studies have shown that PTMs regulate critical proteins in testis development, and their disruptions are implicated in congenital disorders including differences of sex development (DSD), in which sex development is atypical. Despite this, the role and regulation of protein PTM during human testis development remains poorly understood due to limited access to human foetal gonadal tissue, a paucity of large-scale proteomics studies, and a lack of robust of human gonad in vitro models. OBJECTIVE AND RATIONALE: This review aims to provide a comprehensive analysis of validated PTMs affecting proteins critical for testicular development. We discuss PTMs with evidence for a role in normal testis development, and highlight those disrupted in DSD. We review emerging techniques, including proteomic technologies and organ modelling systems that may advance our understanding of PTMs in foetal testis development. We discuss challenges that have restricted the application of these technologies and how overcoming these will significantly improve our understanding of testis development and disease, diagnostics and patient outcomes. SEARCH METHODS: We searched PubMed and the University of Melbourne library for peer-reviewed English-language studies using keywords such as phosphorylation, SUMOylation, acetylation, ubiquitination alongside each protein of interest. PTM sites in proteins involved in testis development were identified using the PhosphoSitePlus database focusing those confirmed in in vitro or animal model studies. ClinVar and the Human Gene Mutation Database were used to identify patient variants that may disrupt PTM sites. OUTCOMES: Our review finds that proteins required for human foetal testis development are subject to extensive PTM. Several PTM sites and PTM-mediated pathways [e.g. MAPK (mitogen-activated protein kinase) pathway] are disrupted in patients with DSD or related conditions. While recent advances in proteomics technologies hold considerable promise, their application to human foetal gonads has been constrained by technical, ethical, and logistical challenges. Encouragingly, emerging high-sensitivity and low-input technologies, alongside stem cell-based approaches, offer viable pathways to overcoming these barriers. WIDER IMPLICATIONS: The relationship between gene regulation, protein expression, and cellular outcome is inherently non-linear, shaped by additional regulatory layers-most notably PTMs. The contribution of PTMs to human testis development in both typical and atypical contexts is a major knowledge gap. Addressing this gap has broad clinical and biological relevance: it may help improve genetic diagnosis or shed light on how proteins or pathways critical for testis development respond to environmental signals-an increasingly pressing question as declining global fertility rates bring testicular function under greater scrutiny. REGISTRATION NUMBER: N/A.

Humans↗
PubMed2007

Laparoscopy for impalpable testis: classification-based management.

BACKGROUND: The undescended testis represents one of the most common disorders of childhood. The authors evaluated the safety and efficacy of laparoscopy for the abdominal testis and present a classification of the laparoscopic diagnostic findings to facilitate decision making. METHODS: Between 2000 and 2005, 95 patients (22 bilateral and 73 unilateral testes, for a total of 117 impalpable testes) with a mean age of 5 years underwent laparoscopy. The testis was managed according to a special classification of the diagnostic findings. Testicular position, size, and viability according to technetium-99m ((99m)Tc) were assessed during the follow-up evaluation. RESULTS: The laparoscopic findings were classified into six types: type 0 (no testis or vanished testis proximal to the internal ring; 9 patients [7.5%]); type 1 (atrophic intracanalicular testis; 6 patients [5.4%], for whom no further intervention was administered); type 2 (testis at the internal ring with looping vas; 15 patients [14.5%], for whom laparoscopic orchiopexy was performed); type 3 (testis at the internal ring without looping of the vas; 29 patients [24.7%], for whom laparoscopic orchiopexy also was performed; type 4 (high abdominal testes; 49 patients [41.9%], with Staged Fowler-Stephens orchiopexy performed for 47 testes and laparoscopic orchidectomy for 2 testes; and type 5 (persistence of Müllerian duct structures [PMDS] or other abnormalities; 7 testes [6%]). After a mean follow-up period of 3 years, the laparoscopic orchiopexy testes were of good size and viable, but four testes (8.7%) were at the neck of the scrotum. The laparoscopically staged Fowler-Stephens orchiopexy group showed atrophy in two testes (4.3%), and all were in the bottom of the scrotum. CONCLUSIONS: Classification of the laparoscopic findings facilitates decision making. Laparoscopic orchiopexy is a natural extension of diagnostic laparoscopy for the intraabdominal testis at the internal ring or that seen peeping from it. Laparoscopically staged Fowler-Stephens orchiopexy is the procedure of choice for the high intraabdominal testis not amenable to the one-stage procedure.

Adult↗
PubMed2002

The management of nonpalpable testis with combined groin exploration and subsequent transinguinal laparoscopy.

PURPOSE: About 20% of all cryptorchid testes are nonpalpable. Although surgical exploration was previously the method of choice for management, laparoscopy from the inferior edge of the umbilicus has been established as a useful method of finding a testis that is nonpalpable. However, conventional subumbilical laparoscopy is unnecessary when the testis or its remnant is located below the internal inguinal ring. We evaluated the efficacy of transinguinal laparoscopy after inguinal exploration for a nonpalpable testis. MATERIALS AND METHODS: While 30 patients each had a unilateral nonpalpable and a contralateral descended palpable testis, 3 had a unilateral nonpalpable and a contralateral undescended palpable testis. When we identified neither a normal testis nor a spermatic cord at exploration of the inguinal canal, we subsequently performed laparoscopic observation through the internal inguinal ring. RESULTS: Of the 30 patients with a unilateral nonpalpable and a contralateral descended testis 8 required transinguinal laparoscopy. However, the procedure was avoided in 22 patients because the testis, its remnant or testicular vessels and vas deferens were detected by inspecting the inguinal region. CONCLUSIONS: Further extended incision into a Pfannenstiel incision was unnecessary in cases of blind ending vas and vessels in the peritoneum with transinguinal laparoscopy. In addition, laparoscopy was avoided in 22 of the 30 children (73.3%) with a unilateral nonpalpable and a contralateral scrotal testis. Our strategy of initial inguinal exploration followed by transinguinal laparoscopy for nonpalpable testis may become a reasonable alternative.

Algorithms↗
PubMed2002

Intra-abdominal testis with loop-like epididymis and intra-canalicular vas and vessels.

A case of intra-abdominal testis with loop-like epididymis and intra-canalicular vas and vessels is presented. A 3-year-old male with left impalpable testis since birth was admitted to our department. Physical examination and ultrasonography were inconclusive. Laparoscopy revealed a small left abdominal testis with surrounding adhesions close to the left-obliterated umbilical artery. The vas deferens and spermatic vessels were entering into the internal inguinal ring. The processus vaginalis was patent. At inguinal exploration the testis was atrophic and the epididymis was loop-like, joining the vas deferens in the inguinal canal. The spermatic vessels continued to the atrophic testis in a loop-like manner. The testis, epididymis and the vas deferens were removed. Histopathological examination of the testis revealed Sertoli cells only. If inguinal exploration had been performed without laparoscopy, the presence of the vas deferens and spermatic vessels in the inguinal canal with the absence of the testis could have been misdiagnosed as vanishing testis. Abdominal testis would thus have been missed, with increased risk of complications, particularly malignancy.

Abnormalities, Multiple↗
PubMed1976

Biochemical actions of follice-stimulating hormone in the sertoli cell of the rat testis.

The sequenc of biochemical events associated with the action of follicle-stimulating hormone (FSH) in the testis has been investigated using a Sertoli cell-enriched testis model system. The Sertoli cell-encriched testis, created by irradiation of male rats in utero, is devoid of germinal elements but contains a normal complement of supportive Sertoli cells. Comparison of the Sertoli cell-enriched testis with normal testis, demonstrates that the two types of testes contain equal numbers of FSH specific receptors, judged by the binding of labeled hormone. In addition, FSH over a concentration range from 6 X 10(-11) to 6 X 10(-9)M will stimulate the production of adenosine 3',5' monophosphate (cAMP) in the Sertoli cell-enriched testis in a manner indistinguishable from that of the normal testis. Incubation of Sertoli cell enriched testis also results in the activation of soluble cAMP-dependent protein kinase. This response to FSH is dependent upon the age of the animal and disappears at about 32 days of age. While sensitivity to the hormone can still be detected in mature Sertoli cell-enriched animals by the addition of the phosphodiesterase inhibitor 1-methyl-3-isobutyl-xanthine, no detectable increase in phosphodiesterase activity is apparent after 30 days of age. Injection of FSH into Sertoli cell-enriched animals results in an increase in total testicular protein synthesis as well as in the production of the Sertoli cell-specific protein, androgen-binding protein within 30 minutes. Furthermore, while hypophysectomy of Sertoli cell-enriched animals result in a decline of the testicular concentration of androgen-binding protein, the injection of FSH will stimulate and maintain the levels of androgen-binding protein in such animals. These results demonstrate that the Sertoli cell-enriched testis is capable of carrying out the sequence of biochemical events previously described for FSH in the normal testis and therefore, suggest that the Sertoli cell is the primary target cell for FSH action.

Age Factors↗
PubMed2002

Ultrasonography is unnecessary in evaluating boys with a nonpalpable testis.

OBJECTIVE: An inguinal sonogram often is obtained in boys with a nonpalpable testis to "localize" the testis, ie, determine whether the testis is present. The results of ultrasonography in boys with a nonpalpable testis were analyzed. METHODS: The records of boys who were referred to a pediatric urology center with a diagnosis of nonpalpable testis and who had undergone inguinal sonography were reviewed. The results of sonography were compared with findings in the office as well as surgical findings. RESULTS: A total of 62 boys who were referred with a diagnosis of a nonpalpable testis and who had undergone a sonogram were reviewed. The sonogram was ordered by the primary care physician in 51 boys (82%) and by a general urologist in 11 cases (18%). The testis was identified by sonography in 12 (18%) of 66 cases, and all were localized to the inguinal canal. Physical examination by a pediatric urologist showed that 6 were in the scrotum and 6 were in the inguinal canal or perineum. Of the 54 testes that were not localized by the sonogram, 33 (61%) were palpable and 21 (39%) were nonpalpable. Of the truly nonpalpable testes, laparoscopy and abdominal/inguinal exploration identified the testis as abdominal in 10 cases and atrophic secondary to spermatic cord torsion in 11 cases. CONCLUSION: Sonography is unnecessary in boys with a nonpalpable testis, because it rarely if ever localizes a true nonpalpable testis, and it does not alter the surgical approach in these patients.

Adolescent↗
PubMed2002

Direct injection of foreign DNA into mouse testis as a possible in vivo gene transfer system via epididymal spermatozoa.

We have attempted to transfect testicular spermatozoa with plasmid DNA by direct injection into testes to obtain transgenic animals [this technique was thus termed "testis-mediated gene transfer (TMGT)"]. When injected males were mated with superovulated females 2 and 3 days after injection, (i) high efficiencies (more than 50%) of gene transmission were achieved in the mid-gestational F0 fetuses, (ii) the copy number of plasmid DNA in the fetuses was estimated to be less than 1 copy per diploid cell, and (iii) overt gene expression was not found in these fetuses. These findings suggest the possibility that plasmid DNA introduced into a testis is rapidly transported to the epididymis and then incorporated by epididymal spermatozoa. The purpose of this study was to elucidate the mechanism of TMGT by introducing trypan blue (TB) or Hoechst 33342 directly into testis. We found that TB is transported to the ducts of the caput epididymis via rete testis within 1 min after testis injection, and TB reached the corpus and cauda epididymis within 2-4 days after injection. Staining of spermatozoa isolated from any portion of epididymis was observed 4 days after injection of a solution containing Hoechst 33342. Injection of enhanced green fluorescent protein (EGFP) expression vector/liposome complex into testis resulted in transfection of epithelial cells of epididymal ducts facing the lumen, although the transfection efficiency appeared to be low. In vivo electroporation toward the caput epididymis immediately after injection of EGFP expression vector into a testis greatly improved the uptake of foreign DNA by the epididymal epithelial cells. PCR analysis using spermatozoa isolated from corpus and cauda epididymis 4 days after injection of a DNA/liposome complex into testis revealed exogenous DNA in these spermatozoa even after treatment with DNase I. These findings indicate that exogenous DNA introduced into tesits is rapidly transported to epididymal ducts via the rete testis and efferent ducts, and then incorporated by epithelial cells of epididymis and epididymal spermatozoa.

Animals↗
PubMed1996

Neonatal polychlorinated biphenyl treatment increases adult testis size and sperm production in the rat.

Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants which decrease serum thyroxine (T4) concentrations. We have previously demonstrated that neonatal hypothyroidism in the rat increases Sertoli cell numbers, adult testis weight, and daily sperm production (DSP). The aim of this study was to determine if neonatal PCB treatment increases adult testis weight and DSP. Treated rats received either Aroclor 1242 or 1254 (0.4-3.2 mg/day), from birth to Day 25 by daily injection; some treated litters also received T4 replacement. Controls received vehicle alone. Tritiated thymidine autoradiography was used to assess Sertoli cell proliferation in 15-day control and Aroclor-treated rats. Serum T4 was measured at 25, 45, and 135 days of age, and serum testosterone, testis weight, DSP, and testicular histology were examined at 135 days. Both Aroclor 1242 and 1254 suppressed serum T4 concentrations; Aroclor 1254 was more potent and long lasting. Testis weight was increased 22 and 13% in rats that received the 1.6 and 3.2 mg/day Aroclor 1242 doses, respectively, while the 0.4 mg/day dose did not produce significant increases. Aroclor 1254 produced significant increases in testis weight of 13 and 23% at the 0.4 and 1.6 mg/day doses, respectively. The 1.6 mg/day Aroclor 1242 and the 0.4 and 1.6 mg/day Aroclor 1254 doses increased DSP by 27, 11, and 42%, respectively; other treatments did not produce significant increases. At 15 days of age, Sertoli cell proliferation was greater in treated rats than in controls. T4 replacement decreased or eliminated the increased testis weight and DSP seen in Aroclor-treated rats. The highest dose of Aroclor 1242 and both doses of Aroclor 1254 decreased adult body weight, while other treatments did not. These results indicate that neonatal PCB treatment increases adult testis weight and DSP in rats. PCBs produce this effect primarily by inducing hypothyroidism, which leads to increased Sertoli cell proliferation, testis weight, and DSP. Thus PCBs, despite inhibitory effects on adult reproductive organs, can paradoxically stimulate increases in adult testis weight and DSP when administered neonatally. These data emphasize the pleiotropic nature of PCB effects and the susceptibility of the developing reproductive system to environmental factors.

Animals↗
PubMed2004

The effects of orchidopexy and orchidectomy on testes of rats subjected to ipsilateral abdominal testis with vas deferens obstruction.

An experimental study was planned to evaluate and compare the effects of orchidopexy and orchidectomy on the testes of rats subjected to unilateral abdominal testis with vas deferens obstruction. Four groups were established. Rats in the control group underwent a sham procedure. While the testis was maintained in the abdomen with the vas deferens ligated for 8 weeks in group 2, rats in groups 3 and 4 underwent orchidopexy or orchidectomy after 4 weeks. Remaining testes were harvested at the end of the 8-week period. Testis and body weights were obtained during harvest. Samples were evaluated through DNA flow cytometry, and percentages of haploid cells were determined. Groups were compared through unpaired t-test, and p-values less than 0.05 were considered significant. All three treatments had decreased testis weight over body weight values of ipsilateral testes. Ipsilateral orchidectomy increased the value among contralateral testes. However, none of the groups had a contralateral testicular value less than the sham-operated group. All three treatments decreased the percentage of haploid cells among ipsilateral testes, but only an abdominal testis was associated with a decrease in the percentage of haploid cells among contralateral testes. Maintaining a testis with an obstructed vas deferens in the abdomen for 8 weeks damages both ipsilateral and contralateral testes. Orchidopexy, while showing amelioration of the ipsilateral testis, spares the contralateral testis as well as orchidectomy. Orchidopexy for an undescended testis with vas deferens obstruction is a rational approach.

Animals↗
PubMed1975

Endogenous steroid production in cellular and subcellular fractions of rat testis after prolonged treatment with gonadotropins.

Steroid production and enzyme activities were examined in preparations of whole testis tissue, isolated interstitial tissue and seminiferous tubules obtained from adult rats with intact pituitaries receiving daily subcutaneous injections of 100 I.U. human chorionic gonadotropin for 5 days and from control animals. After human chorionic gonadotropin administration testosterone concentrations were increased in total homogenates of whole testis tissue, interstitial tissue and seminiferous tubules. The testosterone production from endogenous precursors was enhanced only in total homogenates of whole testis tissue and interstitial tissue obtained from testes of human chorionic gonadotropin-treated rats. The production of testosterone in the corresponding homogenates of isolated seminiferous tubules was very low. The specific activity of 3 beta-hydroxysteroid dehydrogenase was increased in total homogenates of whole testis tissue, isolated interstitial tissue and seminiferous tubules. No effect was observed on the specific activities of marker enzymes such as cytochrome c oxidase, monoamine oxidase, steroid sulfatase and lactate dehydrogenase, whereas the specific activities of carboxyl esterase were decreased in homogenates of whole testis tissue and interstitial tissue. Total activity of monoamine oxidase was increased in homogenates of interstitial tissue of tests from human chorionic gonadotropin treated rats. After the same prolonged human chorionic gonadotropin treatment the concentration of pregnenolone was increased in mitochondrial fractions of whole testis tissue, interstitial tissue and seminiferous tubules, and the amount of protein isolated in the mitochondrial fraction of interstitial tissue increased by 40%. Steroid production (estimated as pregnenolone) from endogenous precusors by mitochondrial fractions of whole testis tissue and interstitial tissue were increased after human chorionic gonadotropin treatment, for whole testis from 580 pmol/mg mitochondrial protein per h to 1420 pmol/mg per h; and for interstitial tissue from 2665 pmol/mg per h to 7050 pmol/mg per h. The production of pregnenolone in mitochondrial fractions obtaine from isolated seminiferous tubules was very low and contributed hardly at all to the total pregnenolone production in mitochondrial fractions of whole testis tissue from normal rats as well as from human chorionic gonadotropin-treated rats.

Animals↗
PubMed1997

Systematic fine-needle aspiration of the testis: correlation to biopsy and results of organ "mapping" for mature sperm in azoospermic men.

OBJECTIVES: This study compares fine-needle aspiration (FNA) and testis biopsy for the ability to detect mature sperm in the testes of azoospermic men. In addition, we introduce the concept of testis "mapping" with FNA and apply it to detect sperm in men with severe testis failure. METHODS: Sixteen patients were evaluated for azoospermia in a university-based infertility clinic. All men had testis biopsies and FNAs from matched testicular sites to assess for the presence of spermatozoa. Adequacy criteria for FNA specimens were strictly defined, and correlative analysis of the two techniques was performed. In addition, a subset of 12 men with nonobstructive azoospermia (NOA) had systematic FNA mapping (more than four FNA sites per testis) to detect mature sperm for potential clinical use. RESULTS: Adequate FNA specimens were obtained in 115 (91.3%) of 126 FNA attempts. Of 34 paired biopsy FNA sites. FNA was seen to be more sensitive than, and equally specific as, testis biopsy for sperm detection. When compared with the biopsy touch imprint, FNA was equally as sensitive and specific. Among men with NOA who underwent FNA mapping, 4 (33%) of 12 had localized "patches" of sperm detected in areas distant from sperm-negative biopsy sites. In 1 case, a pregnancy was achieved with later biopsy and sperm extraction "directed" by previous FNA. CONCLUSIONS: For sperm detection, testis FNA provides equivalent or better information than a testis biopsy. FNA can localize areas of sperm production within the testis and accurately guide sperm extraction procedures in men with NOA.

Biopsy, Needle↗
PubMed2000

Developmentally regulated expression of two transcripts for heme oxygenase-2 with a first exon unique to rat testis: control by corticosterone of the oxygenase protein expression.

Heme oxygenase (HO)-2, the constitutive cognate of oxidative stress inducible HO-1 (HSP32), degrades heme to biliverdin, carbon monoxide, and iron. The highest levels of HO-2 are found in the testis. Previously we identified multiple HO-2 homologous transcripts that differ in size and use three different 5' UTRs that form the untranslated first exon of the gene (referred to as rHO-2, rHO-2-1 and rHO-2-2) and two poly(A) signals. Also, we have characterized a functional glucocorticoid response element (GRE) in the promoter region of rHO-2. In this study, we have examined the structural basis for size heterogeneity of HO-2 transcripts and whether expression of HO-2 at mRNA and protein levels is subject to regulation by corticosterone. Age and tissue-dependence of transcript expression were examined as well. Our data indicate that the remarkable increase in HO-2 mRNA in adult rat testis is due primarily to generation of two HO-2 homologous transcripts of approx. 2.1kb and approx. 1.45kb size that use rHO-2 and are unique to this tissue, and that rHO-2 is not used within other organs. These transcripts are not present in the brain, kidney, thymus, heart, spleen, liver, or in prepubertal 14day old rat testis. The testis-specific transcripts contain all of the coding region exons present in the approx. 1.3kb and approx. 1.9kb transcripts that are common to all organs, including the adult and prepubertal rat testis. Differential use of the poly(A) signals accounts for the difference in size of these two transcripts. Treatment of newborn rats with corticosterone for 5days, starting on day 2 after birth, induced HO-2 protein expression in the testis as detected by Western blotting. In adult rat testis, corticosterone treatment, however, was not an effective regulator of HO-2 transcript populations or levels. The findings suggest that HO-2 levels in the testis are controlled by glucocorticoids; and that developmental and tissue-specific factor(s) determine generation of transcripts unique to the organ. The apparent exclusive use of rHO-2 by the mature testis is consistent with the possibility that HO-2 may play a role in male reproduction.

3' Untranslated Regions↗
PubMed2003

The role of ultrasonographically guided puncture of the human rete testis in the therapeutic management of nonobstructive azoospermia.

We attempted to characterize the cells collected from the rete testis via ultrasonographically guided puncture. Unilateral puncture of the rete testis was performed in nine men with obstructive azoospermia and 51 men with nonobstructive azoospermia. All the aspirated samples from the rete testis were observed via confocal scanning laser microscope and some of them after fluorescent in situ hybridization techniques. Then therapeutic testicular biopsy was performed in the punctured testis of each man. Spermatozoa were found in all rete testis samples and all biopsy samples from obstructed men. Twenty-two nonobstructed men demonstrated absence of spermatozoa in biopsy samples. Twenty-nine nonobstructed men showed spermatozoa in biopsy material and 24 of these men (82%) had demonstrated spermatozoa in rete testis samples. There were no significant differences in fertilization and cleavage rate between intracytoplasmic sperm injection trials using biopsy spermatozoa and rete testis spermatozoa both in obstructed and nonobstructed men. Considering that puncture of the rete testis does not reduce the volume of testicular parenchyma, is less invasive and apparently causes less detrimental effect on testicular vasculature than biopsy, puncture of rete testis is recommended as first line approach for the treatment of azoospermic men. If puncture is negative for spermatozoa in nonobstructed men, biopsy is indicated.

Biopsy↗
PubMed2003

Experimentally induced hypo- and hyper-thyroidism influence on the antioxidant defence system in adult rat testis.

The objective of the present experiment was to study the effect of thyroid hormone on the antioxidant defence system of rat testis. Hypothyroidism induced in rats by 6-n propyl 2-thiouracil (PTU) treatment resulted in a reduction in body weight, seminal vesicle and ventral prostate gland. A further decrease in the weight of seminal vesicle was recorded following administration of T3 to hypothyroid rats. The oxidative stress parameters such as hydrogen peroxide and protein carbonyl content increased in the crude homogenate of testis of hypothyroid rats. T3 administration to hypothyroid rats resulted in no further change in the hydrogen peroxide level but the protein carbonyl content further elevated in the crude homogenate of testis. No significant change was observed in the endogenous lipid peroxidation level of the crude homogenate of testis whereas the FeSO4/ascorbic acid induced lipid peroxidation level decreased in hypothyroid rats and did not change further by T3 administration. Although the reduced glutathione level in the crude homogenate of testis did not change following hypothyroidism, oxidized glutathione level increased. The reduced and oxidized glutathione level decreased and increased, respectively following T3 administration to hypothyroid rats in comparison with PTU-treated rats. Activities of superoxide dismutase and catalase decreased in the post-mitochondrial fraction (PMF) of testis of hypothyroid rats. T3 injection to PTU-treated rats resulted in an elevation in the level of catalase activity only. The activity of glutathione peroxidase in the PMF of testis elevated in the hypothyroid rats and reduced following T3 treatment to hypothyroid rats. The results of the present study suggest that any alteration in the thyroid hormone level in the body affects the antioxidant defence system of testis of adult rats and, thereby, may affect the physiology of testis through oxidative stress.

Animals↗
PubMed2001

Mechanisms involved in contractile differences among cremaster muscles according to localization of testis.

BACKGROUND/PURPOSE: Evidence suggests differences in contractility in cremaster muscles (CM) associated with undescended testis caused by alterations of autonomic innervation. Contractile responses of CM to various pharmacologic agents were evaluated and compared according to the localization of testis. METHODS: Samples of CM from boys with undescended testis or inguinal hernia were obtained. Twitch and tetanic contractions were recorded isometrically at 37 degrees C. Effects of verapamil, isoprenaline, calcitonin gene-related peptide (CGRP), substance P (SP) and N(omega)-nitro-L-arginine (L-NNA) were investigated. Results were compared through 2-way analysis of variance, and P values less than.05 were considered to be different. RESULTS: Verapamil alone significantly (P <.05) decreased contraction amplitudes in CM from both sources; the decrease was more pronounced in CM from boys with inguinal hernia (P <.05). Although isoprenaline increased contraction amplitudes in CM associated with undescended testis (P <.05), CGRP and SP increased contraction amplitudes in CM associated with descended testis (P <.05). L-NNA increased contraction amplitudes in both groups (P <.05). The decrease of contraction amplitudes after verapamil displayed a similar pattern after isoprenaline, SP, and L-NNA. Verapamil-induced contractility decrease was more pronounced after CGRP in both groups (P <.05). CONCLUSIONS: Sensitivity of CM to verapamil differs according to localization of testis. Isoprenaline enhances contractility by stimulating Na(+)-K(+)ATPase in undescended testis without altering voltage-sensitive channel sensitivity to verapamil. CGRP and SP increase contractility in inguinal hernia, and CGRP increases the sensitivity of voltage-sensitive Ca(2+) channels to verapamil in CM from both groups. Nitric Oxide (NO) exerts inhibitory action on CM contractility, and it is less pronounced in undescended testis. These differences may contribute to pathophysiology of undescended testis.

Adult↗
PubMed1999

Radiation-induced apoptosis and gene expression in neonatal kidney and testis with and without protein synthesis inhibition.

PURPOSE: To analyse the incidence of radiation-induced apoptosis, expression of two apoptosis-related genes, Bcl-2 and p53, and post-radiation levels of cell proliferation in the neonatal rat (4-5 days old) kidney and testis. MATERIALS AND METHODS: Apoptosis was quantified in control or treated kidney or testis at 2, 4, 6, 8 and 24h after 5 Gy of whole body X-irradiation (n=4 per group). Morphology (light and electron microscopy) and DNA gel electrophoresis were used to assess apoptosis. Temporal and spatial expression of Bcl-2 or p53 were analysed using immunohistochemistry. Administration of cycloheximide (1.5mg/kg) was used to determine whether new protein synthesis had a role in induction of apoptosis. Tritiated thymidine uptake and autoradiography were used to indicate alterations in cell proliferation (radiolabel administered 1 h prior to tissue collection) or S-phase cells undergoing radiation-induced apoptosis (radiolabel administered 1 h prior to irradiation). RESULTS: Apoptosis peaked at 4 h in the testis and 6 h in the kidney and was significantly higher in the renal nephrogenic zone than in the testis (p<0.05). Mitosis was almost completely negated after irradiation in both tissues. A higher proportion (almost fivefold) of the apoptotic cells died in S phase in the kidney than in the testis. Cycloheximide negated induction of apoptosis in the kidney, and markedly decreased apoptosis in the testis. Bcl-2 expression was highest in the differentiated zone of control kidneys and increased after irradiation in the nephrogenic zone, particularly near foci of apoptosis in developing nephrons. In the control testis, Sertoli cells had moderate expression of Bcl-2. After irradiation, there was complete absence of Bcl-2 expression in apoptotic Sertoli cells, with surviving cells increasing Bcl-2 expression. Irradiated kidney had more intense nuclear p53 expression compared with controls. In the testis, p53 that was present in controls continued to be expressed in surviving cells but not apoptotic cells in radiation-treated animals. CONCLUSIONS: Unique differences can be identified between the incidence and biomolecular control of radiation-induced apoptosis in the normal neonatal kidney and testis. These results may find application for minimizing damage to these normal neonatal tissues in the development of, for example, cancer treatment regimens.

Animals↗
PubMed1998

Alternative splicing of exons in the alpha1 subunit of the rat testis L-type voltage-dependent calcium channel generates germ line-specific dihydropyridine binding sites.

Cell-specific isoforms of the alpha1 subunit of the L-type voltage-dependent calcium channel (VDCC) have unique pharmacological reactivities. Prior sequence analysis of nucleotide bases 3908-6077 of the VDCC alpha1 subunit expressed in rat testis differed from cardiac sequences only in a 84 base pair region corresponding to exons 31/32 encoding a putative dihydropyridine binding region. We now report that sequence analysis of bases 3048-3936 identifies a second difference between the rat testis and rat cardiac alpha1 sequence in a 60 base pair region corresponding to exons 21/22 and encoding another putative dihydropyridine binding site. Variable VDCC exons 21/22 and 31/32 and their linking introns were sequenced using genomic DNA from rat lung as template, providing evidence that the rat testis and cardiac alpha1 isoforms are products of the same gene. Reverse transcription in-situ polymerase chain reaction (PCR) with frozen sections of rat testis was carried out with primers identifying the testis-specific exon 32 of the VDCC alpha1 subunit. PCR products were confined to seminiferous tubules and were associated with the germ cell lineage from Type A spermatogonia to mature spermatozoa. Close coupling of testis alpha1 VDCC gene transcription and translation was established by in-situ immunolabelling of serial frozen sections with a monoclonal antibody (IIF7) directed against epitopes on rabbit skeletal muscle L-type VDCC alpha1. Western blot analysis of rat proteins extracted from heart, skeletal muscle, testis and spermatozoa which were reactive with the IIF7 antibody detected primarily 175-220 kDa proteins in the size range of VDCC. These data unequivocally demonstrate that an L-type VDCC is expressed in rat testis and that VDCC isoforms from rat testis and heart differ in deduced amino acid composition in and around potential binding sites for calcium channel blocking drugs such as the dihydropyridines.

Alternative Splicing↗
PubMed1987

Purification and properties of porcine testis acylphosphatase.

Acylphosphatase has been purified from porcine testis and its properties were compared with those of porcine skeletal muscle acylphosphatase. The molecular weight of the testis enzyme was found to be 10,600, similar to that of porcine skeletal muscle acylphosphatase, on sedimentation equilibrium analysis. The specific activity of the testis enzyme was 10,800 mumol/min/mg at 25 degrees C with benzoyl phosphate as substrate, i.e., higher than that of the muscle enzyme, 7,200 mumol/min/mg, under the same conditions. The pI of the testis enzyme was 8.3, i.e., lower than that of the muscle enzyme, 10.6. There were marked differences in the amino acid compositions of the two enzymes. In particular two histidine residues were present in the testis enzyme but none were present in the muscle enzyme, and no cysteine residue was present in the testis enzyme but one was present in the muscle enzyme. The carboxyl terminal amino acid of the testis enzyme seemed to be lysine, while that of the muscle enzyme is tyrosine. The peptide maps of the testis and muscle enzymes indicated considerable differences in the amino acid sequences of the two enzymes. Differences in the antigenic structures of the two enzymes were demonstrated on enzyme linked immunoassaying and double immunodiffusion. These results indicate that the porcine testis acylphosphatase is an isozyme different from the porcine skeletal muscle acylphosphatase.

Acid Anhydride Hydrolases↗