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PubMed2003

Expression and localization of Smad1, Smad2 and Smad4 proteins in rat testis during postnatal development.

AIM: To study the expression and regulation of Smad1, Smad2 and Smad4 proteins (intracellular signaling molecules of transforming growth factor-b family) in rat testis during postnatal development. METHODS: The whole testes were collected from SD rats aged 3, 7, 14, 28 and 90 (adult) days. The cellular localization and developmental changes were examined by immunohistochemistry ABC method with the glucose oxidase-DAB-nickel enhancement technique. Quantitative analysis of the immunostaining was made by the image analysis system. The Smads proteins coexistence in the adult rat testis was tested by the double immune staining for CD14-Smad4 and Smad2-Smad4. The protein expression of Smad during rat testicular development was examined by means of Western blots. RESULTS: Smad1, Smad2 and Smad4 were present throughout testicular development. The immunostaining of Smad1 and Smad2 were present in spermatogenic cells. A positive immunoreactivity was located at the cytoplasm, but the nucleus was negative. Smad1 was immunolocalized at the d14, d28 and adult testes, while Smad2, at the d7, d14, d28 and adult testis. There was positive immunoreaction in the Sertoli cells and Leydig cells as well. The immunolocalization of Smad4 was exclusively at the cytoplasm of Leydig cells and the nuclei were negative throughout the testicular development. No expression was detected in the germ cells. The results of image and statistical analysis showed that generally the expression of Smad1, Smad2 and Smad4 in the testis tended to increase gradually with the growth of the rat. CONCLUSION: The present data provide direct evidences for the molecular mechanism of TGF-bgr action in rat testes during postnatal development and spermatogenesis.

Animals↗
PubMed2003

Targeted expression of human FSH receptor Asp567Gly mutant mRNA in testis of transgenic mice: role of human FSH receptor promoter.

AIM: To specifically express the Asp567Gly human follicle-stimulating hormone receptor (FSHR) under the control of its promoter to evaluate the phenotypic consequences in the presence of normal pituitary function. METHODS: We produced transgenic mice overexpressing the Asp567Gly human FSHR under the control of a 1.5kb 5'-flanking region fragment of its promoter. RESULTS: Mice were phenotypically normal and fertile. In males, mRNA could be detected in the testis and the brain, indicating that the 1.5kb promoter fragment drives expression not only in the gonads. The testis weight/body weight ratio and the testosterone levels in transgenic and non-transgenic littermates were similar. By in situ hybridisation we found that the transgenic FSHR was highly expressed in Sertoli cells, spermatocytes and round spermatids. However, a radioligand receptor assay failed to show a significant difference in total FSHR binding sites in testis homogenates of transgenic and wild type animals, suggesting that the transgenic FSHR is probably not translated into functional receptor protein. CONCLUSION: A 1.5kb 5'-region of the human FSHR drives mRNA expression of the transgene in the testis but leads to ectopic expression in germ cells and in the brain. No phenotypic consequences could be documented due to the lack of protein expression.

Animals↗
PubMed2004

[Expression changes of tau protein and microtubule-associated proteins in testis intersitium of rats].

OBJECTIVE: To examine the expression of the tau protein and microtubule-associated proteins in the testis interstitium of aged and young rats. METHODS: Sprague-Dawley male rats were divided into a young group(6 months, n = 10) and an aged group(28 months, n = 10). The two steps immunohistochemistry method with the antibody against tau protein and MAP alpha was performed on the testis tissues of the rats. RESULTS: The results showed that the immunoreactive cells of tau protein of the testis interstilial of the aged rats obviously increased(P < 0.001) than those of the young, while the immunoreactive cells of the microtubule-associated proteins decreased(P < 0.01) in the aged. CONCLUSION: The changes in the expression of the tau protein and microtubule-associated proteins may be related to the aging process of the testis.

Aging↗
PubMed2004

Developmental changes in inhibin-alpha gene expression in the mouse testis.

Inhibin is a gonadal hormone which is composed of an alpha-subunit and one of two related beta-subunits (betaA, betaB). Inhibin is important for pituitary FSH regulation, normal follicle development and maintenance of the estrous cycle in the female, whereas the role of inhibin in the male is less clear. Thus, we examined the expression of the inhibin-alpha gene in testis during sexual maturation in male mice, to try to gain insight into its functions in the male. Male mice of the ICR strain attained fertility at 6 weeks of age, and histological analysis revealed that a functional testis was formed, with seminiferous tubules which contain mature sperm and with an abundant population of Leydig cells. Parallel with this sexual maturation, inhibin-alpha subunit protein synthesis increased, whereas synthesis of the activin betaA and activin betaB followed with a delayed time course. Inhibin-alpha mRNA also increased during this critical period, and this corresponded to a change in the methylation status of the inhibin-alpha gene. Taken together, our data reveal that activation of inhibin-alpha gene during testis development correlated with the histological maturation of the testis and the acquisition of fertility in male mice.

Animals↗
PubMedDate not supplied

Lectin-binding sites in newborn human testis.

The present investigation has been performed to obtain a complete distributional map of the oligosaccharidic component of the glycoconjugates in the testis of the human newborn. For this purpose seven eight HRP-conjugated lectins (SBA, DBA, PNA, WGA, OOA, UEAI, LTA and ConA) along with enzymatic treatments, were used. The Sertoli cells were characterized by the same sugar residues detected in the testes of adult healthy subject. Beta-N-acetyl-D-galactosamine in the Leydig's cells and a-N- acetyl-D-galactosamine anomer in the spermatogonia seem to be an unique feature of the newborn testis. For this fact SBA and DBA could be considered markers respectively of the Leydig's cells and of the spermatogonia in the newborn testis. Differences in lectin binding, between the newborn and the adult testis as the interstitial tissue and the endothelial cells of the capillary vessels are reported.

Binding Sites↗
PubMed2004

[Perineal ectopic testis: report of four paediatric cases].

Perineal ectopic testis is a rare congenital malformation in which the testis is abnormally situated between the penoscrotal raphe and the genitofemoral fold. The authors report four new cases in children aged 2 months, 6 months, 2 years and 5 years. The abnormality was associated with an inguinal hernia in one case. The diagnosis was based on the presence of an empty scrotum or perineal swelling. Treatment, via an inguinal incision, consisted of orchidopexy in a dartos pouch with a favourable course in every case. The aetiopathogenesis of perineal ectopic testis is controversial. It can be easily diagnosed by palpation of the testis in the perineal region. Orchidopexy in a dartos pouch must be performed early, and does not raise any particular problems because of the sufficient length of the spermatic cord. The functional prognosis, always difficult to define, appears to be identical to that of other sites.

Child, Preschool↗
PubMed2005

[The changes of IGF-I in testis and epididymis on a rat model with oligozoospermia/azoospermia induced by cyclophosphamide].

OBJECTIVE: To evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide. METHODS: Ninety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis. RESULTS: The levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced. CONCLUSION: In the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.

Animals↗
PubMed2005

[Testicular torsion after previous fixation of the testis].

Testicular torsion (TT) is caused by rotation of the testis. If not treated acutely, TT causes ischaemic necrosis. As the anatomic variations responsible for the torsion are likely to be bilateral, the opposite testis should also be fixed. We report on a case where a patient was admitted to hospital with clinical signs of TT. Two years earlier the patient had undergone fixation of the testis in an operation for TT on the opposite side. Patients with clinical signs of TT should be explored even if both testis have previously been fixed.

Adolescent↗
PubMed2005

Effect of estradiol on expression of cytoskeletal proteins during spermatogenesis in testis of sexually mature rats.

Earlier studies had shown that long term treatment with estradiol arrests spermatogenesis in adult male rats, at a dose of 0.1 mg/kg/day. The present study was therefore undertaken to ascertain the causes underlying the reduction in sperm counts by administering estradiol for a short term, at the dose of 0.1 mg/kg/day. Estradiol valerate was injected at a dose of 0.1 mg/kg/day, for a period of 10 days to one group of adult male rats, which were administered saline for 12 days prior to estradiol injection, and sacrificed after 22 days. The control group was administered saline for 22 days. The sera were analyzed for testosterone and FSH levels. One testis of each male was immersion fixed for histology, and for immunohistochemistry of two testicular cytoskeletal proteins, vimentin and vinculin. The contralateral testes were used for analysis of vimentin and vinculin gene expression by reverse transcriptase polymerase chain reaction (RTPCR) and western blotting. Another group exposed to estradiol for 10 days was injected with bromodeoxyuridine (BrdU), at a dose of 100 mg/kg/day, to ascertain the effect on germ cell proliferation, and sacrificed 12 days later, while estradiol treatment was continued till sacrifice. BrdU, at a dose of 100 mg/kg/day was injected i.p. to a group of control rats treated with saline for 10 days, and sacrificed 12 days later. The testes from both groups were immersion fixed for immunohistochemical detection of BrdU. Histology of estradiol treated testis showed predominance of tubules with round spermatids with accumulation of lipid droplets in Sertoli cell cytoplasm and decreased cell height, whereas controls showed elongating spermatids. BrdU immunolocalization in the testis, irrespective of treatment, indicated its incorporation in deoxyribonucleic acid (DNA) suggesting that estradiol sustained germ cell proliferation. Both vimentin and vinculin could be immunolocalized to the testis. The testicular levels of vimentin and vinculin, quantified after western blotting, were unaffected. The testicular expression of vimentin and vinculin seen by RTPCR was also unaffected. The study suggested that estradiol induced reduction in sperm counts was not due to adverse effects on proliferation. The observed predominance of seminiferous tubules showing round spermatids, accumulation of lipid droplets as compared to controls suggested that reduction in elongated spermatids occurred through reduced spermiation and phagocytosis. The study also suggested that reduction in Sertoli cell height after short-term estradiol treatment was not due to reduced expression of vimentin and vinculin, which could be maintained by estradiol. However, reduction in Sertoli cell height could have been due to suppression of FSH and testosterone, implicated in the polymerization of vimentin and organization of vinculin, two cytoskeletal proteins involved in inter-Sertoli or Sertoli-germ cell junctions. The study suggested that disorganization of Sertoli cell cytoskeleton and reduction in the volume of Sertoli cells could be an important factor for reduced efficiency of spermatogenesis after exposure to estrogenic molecules.

Animals↗
PubMed2005

Neonatal hypothyroidism-induced changes in rat testis size, dependence on temperature.

OBJECTIVE: Effects of transient neonatal hypothyroidism (HPOT) on adult testis size and serum hormone profiles were evaluated in the Charles foster strain of rats, maintained under the temperature of 21 degrees C (HPOTL) or of 34 degrees C (HPOTH). METHODS: Hypothyroidism was induced in suckling pups in preweanling period (1-21 days), by administering 0.1% 6-propyl-2-thiouracil in drinking water to mothers. Body mass and testis characteristics, the steroidogenic potential [measured by histochemical localization of 3alpha-, 3beta- and 17beta-hydroxysteroid dehydrogenases (HSDHs)] and serum concentrations of thyroid-stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), luteinizing hormone (LH), testosterone (T) and corticosterone (Cort) were evaluated on the 35th, 45th, 60th and 90th day of age. RESULTS: The HPOTH rats showed lower testis masses, while the HPOTL rats showed higher testis masses and lower body masses after 90 days. Histologically, the testes of the HPOTH rats demonstrated increased germ cell degeneration after 35 and 45 days and reduced tubular size, germ cell numbers and sperm density after 90 days. In contrast, the testes of the HPOTL rats showed reduced tubular diameters after 35 and 45 days, and increased tubular diameters, germ cell numbers and sperm density after 90 days. Serum TSH, T3, T4, LH and T concentrations and 3beta- and 17beta-HSDH activities were reduced in both groups of the HPOT rats after 35 and 45 days. Intratubular steroidogenesis and a prominent triangle up5 pathway were also found in the HPOT animals. CONCLUSION: The temperature has a definite influence on the thyroid hormone action, postnatal growth and function of rat testes.

Age Factors↗
PubMed2006

Humoral immune responses to testis antigens in sera from patients with prostate cancer.

Tumor vaccines represent one type of molecularly targeted therapy being investigated for the treatment of prostate cancer. Although many prostate-specific proteins are being tested as target antigens for prostate cancer vaccines, most are not natural targets of an immune response in patients with cancer. Using sera from cancer patients, several research groups have identified a large family of immunologically recognized proteins whose expression is normally confined to immune-privileged testis tissue but which may be expressed in cancers of different histological origins. These proteins, so-called cancer-testis (CT) antigens, are appealing targets for immune-based therapies because they are essentially tumor-restricted antigens and there is less risk of preexisting immune tolerance. In addition, specifically targeting these proteins by means of vaccines should reduce the risk of potential autoimmune reactions to normal tissues. In the current study, we hypothesize that prostate CT antigens can be identified using a SEREX screening method with sera from patients with prostate cancer and probing with a human testis cDNA expression library. We have identified several potential prostate cancer antigens with predominantly testis-specific expression in normal tissues, including MAD-CT-1 (protamine 2) and MAD-CT-2. Each was independently identified from different subjects with prostate cancer. Antigens identified by these studies can be investigated further as potential prostate cancer tumor antigens.

Adult↗
PubMed2005

Cystic dysplasia of the rete testis associated to cryptorchidism: a case report.

A case of cystic dysplasia of the rete testis (CDRT) is reported. Nine months after right orchidopexy, a 10-year-old boy presented a palpable mass in the ipsilateral hemi-scrotum. Preoperative ultrasound showed a cystic lesion involving almost the whole testis. Radical orchidectomy was performed and at gross examination a hilar multiloculated cystic lesion was confirmed; microscopic examination showed cystic spaces lined by benign appearing monostratified epithelium with atrophy of the surrounding testicular parenchyma. These findings were consistent with the diagnosis of cystic dysplasia of the rete testis. Immunohistochemical investigation evidenced a pattern similar to that of a normal rete testis. Renal ultrasound and DTPA scintigraphy performed after pathological diagnosis documented the absence of the ipsilateral kidney. CDRT is a rare cause of scrotal swelling in the pediatric patient. When diagnosis is suspected, based on clinical and sonographic findings, conservative surgery may be considered.

Abnormalities, Multiple↗
PubMed2006

[HCG in the treatment of cryptorchidism. The effect of age and position of the testis].

In a recent study, the effect of human chorionic gonadotropin (HCG) in the treatment of children with cryptorchidism aged less than four years was questioned. The purpose of the present study was to determine, on the basis of a retrospective examination of patients' records, the effect of HCG treatment, whether outcomes are age-dependent and whether the effects of treatment are related to the position of the testis. Patients diagnosed with undescended testis who had been treated at the Department of Urology at Gentofte Hospital (Copenhagen) in the period from 1 November 1998 to 31 May 2003 were identified. Patients who had been treated with HCG were included in the sample. Boys who at an earlier stage had been operated on in the inguinal or scrotal region, or who had suffered from inguinal hernia, were excluded, and clinical retractile testes were separated from true undescended testes. The criterion of success for efficient treatment was complete descent to the bottom of the scrotum. In the 306 records examined, 121 patients met the inclusion criteria. In total, there were 170 undescended testes. The patients' median age at time of treatment was 3.6 years. The overall success rate was 34.7%. The success rates in the individual age categories were: 1-2 years, 36.7%; 3-4 years, 33.4%; and 5-13 years, 35%. The position of the testis at the beginning of treatment showed that the lower the pretreatment position, the better the success rate. The overall success of HCG treatment of cryptorchidism was 34.7%. No age dependency of HCG effects was found, but the position of the testis before treatment influenced the success rate.

Age Factors↗
PubMed2006

[Expression and role of nitric oxide synthase in the testis and epididymis of Macaca fascicularis].

OBJECTIVE: To investigate the expression and the role of nitric oxide synthase (NOS) in the testis and epididymis of macaca fascicularis. METHODS: The immunohistochemical ABC method was used to observe the localization of nitric oxide synthase in the testis and epididymis of the macaca fascicularis. RESULTS: (1) nNOS immunoreactivity was found in the spermatogenic cells of seminiferous tubules, the epithelia of epididymal efferent ducts, sperm and the endothelia of blood vessels; (2) iNOS was expressed in the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels; (3) eNOS immunoreactivity was detected in the interstitial cells of the testis, the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels. CONCLUSION: NOS is extensively expressed in the testis and epididymis of the macaca fascicularis and it may play an important role in such processes as spermatogenesis, sperm maturation and testosterone secretion.

Animals↗
PubMed2006

[Regulatory effect of Wenyang Whengling Decoction on Smads expressions in testis of sterile rats with Shen-yang deficiency].

OBJECTIVE: To study the localization and distribution of expressions of Smads (mother against dpp), the intracellular signal transduction molecules in the transforming growth factor-beta (TGF-beta) family, in the testis of male sterile rats with Shen-yang deficiency induced by adenine and to observe the effect of Wenyang Shengjing Decoction (WSD) on these expressions. METHODS: Rats were randomly divided into the control group, the model group and the WSD group. Localization and distribution of Smad 1, Smad 2 and Smad 4 expressions were detected by immunohistochemistry SABC and semi-quantitative RT-PCR and analyzed statistically by image analysis system; the contents of testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were detected by radioimmunoassay; and the weights of body, testis and epididymis, as well as sperm number of rats were also measured. RESULTS: Smad 1 and Smad 2 were expressed in cytoplasm of all levels of spermatogenic cells in rats' testis with their positive immuno-responsive substance locating in the cytoplasm, and positive Smad 2 expression could also be found in cytoplasm of Sertoli's cell, but both of them showed negative response in Leydig's cell; Smad 4 was positively expressed in cytoplasm of Leydig's cell but showed negative response in spermatogenic cell and Sertoli's cell. Compared with the normal control, Smad 1 expression was lower (P < 0.05), but Smad 2 and Smad 4 were higher in the model group (both P < 0.05), these abnormal changes could be reversed by WSD treatment (all P < 0.05). Compared with the normal control group, the body weight, sperm number and serum T level were lower, and levels of FSH and LH were higher (all P < 0.05) in the model group, which could all be improved by WSD (P < 0.05); the weights of testis and epididymis were unchanged in all groups (P > 0.05). CONCLUSION: WSD could not only increase the sperm number through elevating serum T level and decreasing the levels of FSH and LH, but also by way of regulating Smads genes expression to adjust the levels of sex hormones, promote the production of sperm directly or indirectly, so as to treat male infertility.

Animals↗
PubMedDate not supplied

Atrial natriuretic peptide precursor material in rat testis.

Atrial natriuretic peptide (ANP) has recently been suggested to play a role in the regulation of male reproduction. Specifically, it has been shown to affect steroidogenesis in Leydig cells. The data presented here demonstrate that the rat testis also contains immunoreactive, ANP-like material (IR-ANP). Characterization of this material was conducted by high-pressure liquid chromatography (HPLC) analysis employing two different antisera to recognize either the C- or the N-terminals of the ANP prohormone (ANP 1-126). The IR-ANP corresponded to the 15 kd ANP precursor molecule (ANP 1-126), indicating the synthesis of ANP within the testis. This was supported by the detection of ANP messenger RNA transcripts (Northern blot hybridization) in the testis. These data indicate that ANP is expressed in the rat testis and support the notion that it may function in a paracrine and/or autocrine fashion in this organ.

Animals↗
PubMed1977

[Long-term study on the influence of running exercise, food restriction, running exercise + food restriction and of parenterally applicated testis cells on age-parameters of the rat (author's transl)].

Preliminary results of a long-term study on 1100 male Sprague-Dawley rats are presented, showing the influence of running exercise, of restricted diet, of both of these together and of the s.c. application of lypholized testis cells. Up to now animals aged 9, 15 and 24 mths were investigated. The test-animals were exposed to the experimental conditions from their 6th month of life. The running exercise was carried out on a treadmill (30 m at a speed of 25 m/min, horizontal) 5 days a week. A food restriction of about 20% was achieved by 2 fasting-days a week. The lyophilized testis cells were injected for the first time at an age of 9 mths and afterwards in 6 mths intervals. Between the injection and the assessment of the age-parameters there was an interval of 6 mths. S o fare the following parameters of the comprehensive test-program have been evaluated: 1) running performance in m (treadmill, 20 m/min, ascent 15 degrees), 2) motor activity (Animex Activity Meter), 3) chemical contraction-relaxation of tail-tendon-fibers, 4) total lipids and total cholesterol in the plasma. The results obtained so far show that a mild regular training, moderate food restriction and the s.c. application of testis-cells are able to cause a significant shift in the dirction of a younger biological age in at least some of the age parameters. The action of the testis-cells seems to affect most of the age parameters but shows the tendency to be more distinct at a higher age. More concrete statements about the influence on the biological age or the vitality will only be possible after the multivariate analysis of the results.

Age Factors↗
PubMed1990

A distinct human testis and brain mu-class glutathione S-transferase. Molecular cloning and characterization of a form present even in individuals lacking hepatic type mu isoenzymes.

mu-Class glutathione S-transferases (GSTs) were identified in all 13 human testes and 28 brains examined; even subjects whose livers were devoid of mu-GSTs expressed extrahepatic GSTs of this class. Testes and brains from individuals with mu-class GSTs in their livers had additional forms that also reflected the liver phenotypes. An isoenzyme with an isoelectric point of 5.2, which was a major GST in testis and present as well in cerebral cortex but not detected in any livers, was identified and purified. Sequence analysis of peptides derived by cleavage of the testicular mu-class GST by Achromobacter protease I revealed distinct aspects of primary structure not found previously in any mammalian mu-class GSTs. These unique features included a blocked and extended amino terminus and 3 additional residues (Pro-Val-Cys) at the carboxyl terminus. This structure was confirmed by molecular cloning and sequencing of cDNAs derived from human testis and brain libraries. In the coding region the mRNA of the brain-testis mu-class GST was 75% homologous with that of the liver form, and its 3'-untranslated sequence was mostly divergent, indicating that it is the product of a separate gene. Distinct catalytic and structural properties of the testis-brain mu-class GSTs suggest that these GSTs may be uniquely involved in blood-barrier functions common to both organs.

Adolescent↗