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PubMed1984

The activity and biosynthesis of cholesterol side-chain cleavage enzyme in cultured immature pig testis cells.

An IgG fraction from antiserum raised against cholesterol side-chain cleavage cytochrome P-450 (cytochrome P-450scc) purified from bovine adrenocortical mitochondria cross-reacted with immature pig testis cytochrome P-450scc in an Ouchterlony double diffusion system. This property of the IgG fraction was utilized for the immunoisolation of cytochrome P-450scc of immature pig testis cells in culture. The molecular weight of the immunoisolate from [35S]methionine-labeled pig testis Leydig cells was similar to that of purified bovine adrenocortical cytochrome P-450scc (49 000). The testis iron-sulfur protein similarly immunoisolated using an IgG fraction from antiserum raised against adrenodoxin purified from bovine adrenocortical mitochondria was also of molecular weight similar to that of adrenodoxin (13 000). The rates of synthesis of cytochrome P-450scc and testis iron-sulfur protein in immature pig testis cells in culture, when incubated with hCG (100 mU/ml) for 48 h, were 3- and 6-fold greater, respectively, than those in cells incubated in the absence of hCG. This result is suggestive that the synthesis of cytochrome P-450scc and testis iron-sulfur protein of immature pig testis tissue is stimulated by gonadotropin.

Animals↗

PubMed1991

The effect of dietary vitamin E and beta-carotene on oxidation processes in the rat testis.

The effects of dietary vitamin E and beta-carotene were studied on enzymes involved in arachidonic acid metabolism and other related enzymes in the rat testis. Groups of rats were fed various soybean oil-based semi purified diets. Group 1 was fed a vitamin E-supplemented diet (+E - beta); Group 2 was fed a beta-carotene-supplemented diet (-E + beta); Group 3, the control group (-E - beta) was fed a vitamin E-deficient diet; and Group 4, the standard diet group (S), was fed vitamin E plus beta-carotene-standard diet. Soybean oxidized oil was added to the three diet groups - (+E - beta), (- E + beta) and (- E - beta), whereas the diet of S group contained non-oxidized oil. After 8 weeks rats were killed, blood and testis samples were collected for biochemical determinations. Vitamin E deficiency caused significant increase in testis thiobarbituric acid value and activities of testis NADPH oxidase, testis 15-lipoxygenase and in plasma pyruvate kinase. In contrast, significant decreases were observed in activity of testis prostaglandin synthetase, compared with antioxidant-supplemented diet groups. We also found a significant increase in 15-lipoxygenase activity in (- E + beta) diet group, compared with (- E - beta) diet group. Fatty acid analysis of testis parenchyma indicated decrease in palmitate (16:0) and arachidonate (20:4(n - 6)), and increase in oleate (18:1(n-6)) linoleate (18:2(n - 6)) and linolenate (18:3(n - 3)), when compared (-E - beta) diet group with vitamin E-supplemented diet groups. The results suggest that dietary vitamin E has a role in both enzymatic and non-enzymatic peroxidation of polyunsaturated fatty acids in the testis.

Animals↗

PubMed1989

Sulfation in male reproductive organs. Bull and boar testis phenol sulfotransferases.

Phenol sulfotransferases (PST) from bull and boar testis were partially purified and characterized. A single form of PST adsorbed on DEAE-cellulose was found in the bull testis, whereas from boar testis two different peaks of PST activity were separated. The bull testis PST and both boar testis enzymes were active with p-nitrophenol and adrenalin. They all showed higher affinity to pNP than to adrenalin and were inhibited by these substrates at higher concentrations. Their optimal pH was at 8.5. Bull testis PST and boar PST II which were adsorbed on DEAE-cellulose were thermostable, whereas boar PST I was thermolabile. Those three PST forms differed in sensitivity to 2,6-dichloro-4-nitrophenol (DCNP), N-ethyl maleimide (NEM), iodoacetamide (IAA) and phenylglyoxal (PG). Bull and boar PST II were more rapidly inactivated in the presence of DCNP than boar PST I. In the presence of NEM, the--SH groups reagent, the bull phenol sulfotransferase and boar PST I lost their activity, whereas the activity of boar PST increased. Also iodoacetamide, another--SH group modificator, raised boar PST II activity and decreased boar PST I activity. DTT, which protects thiol groups, had an opposite effect on the enzymes studied than NEM. Phenylglyoxal, a reagent specific for arginine residues inhibited bull testis PST and both boar phenol sulfotransferases. Substrate protection experiments were also performed to determine the localization of reactive groups in bull and boar testis phenol sulfotransferases.

Animals↗

PubMed2006

The effect of alpha-tocopherol on lipid peroxidation of microsomes and mitochondria from rat testis.

The testis is a remarkably active metabolic organ; hence it is suitable not only for studies of lipid metabolism in the organ itself but also for the study of lipid peroxidation processes in general. The content of fatty acids in testis is high with a prevalence of polyunsaturated fatty acids (PUFA) which renders this tissue very susceptible to lipid peroxidation. Studies were carried out to evaluate the effect of alpha-tocopherol in vitro on ascorbate-Fe(++) lipid peroxidation of rat testis microsomes and mitochondria. Chemiluminescence and fatty acid composition were used as an index of the oxidative destruction of lipids. Special attention was paid to the changes produced on the highly PUFA [C20:4 n6] and [C22:5 n6]. Lipid peroxidation of testis microsomes or mitochondria induced a significant decrease of both fatty acids. Total chemiluminescence was similar in both kinds of organelles when the peroxidized without (control) and with ascorbate-Fe(++) (peroxidized) groups were compared. Arachidonic acid was protected more efficiently than docosapentaenoic acid at all alpha-tocopherol concentrations tested when rat testis microsomes or mitochondria were incubated with ascorbate-Fe(++). The maximal percentage of inhibition in both organelles was approximately 70%; corresponding to an alpha-tocopherol concentration between 1 and 0.25 mM. IC50 values from the inhibition of alpha-tocopherol on the chemiluminescence were higher in microsomes (0.144 mM) than mitochondria (0.078 mM). The protective effect observed by alpha-tocopherol in rat testis mitochondria was higher compared with microsomes, associated with the higher amount of [C20:4 n6]+[C22:5 n6] in microsomes that in mitochondria. It is proposed that the vulnerability to lipid peroxidation of rat testis microsomes and mitochondria is different because of the different proportion of PUFA in these organelles The peroxidizability index (PI) was positively correlated with the level of long chain fatty acids. The results demonstrated the protective effect of alpha-tocopherol on lipid peroxidation in microsomes and mitochondria from rat testis.

Animals↗

PubMed1997

The retractile testis can be a cause of adult infertility.

OBJECTIVE: To determine the functional state of the testes of young adults treated for small and hypotrophic retractile testis at prepubertal age by orchiopexy and/or hormonal therapy and the functional state of the testes of adults with retractile testis. DESIGN: Spermiogram and transmission electron microscopy (TEM) study of the semen. Light microscopic and TEM studies of semen and testicular biopsies of adult with retractile testis were performed. SETTING: Division of Pediatric Surgery in an academic environment. PATIENT(S): Thirty-eight young adults (mean age, 18 years) treated for retractile testis at prepubertal age and seven adults (mean age, 28 years) with retractile testis. INTERVENTION(S): Two cycles of hCG, followed by surgical therapy (orchiopexy) when unsuccessful. MAIN OUTCOME MEASURE(S): Fertility of young adults treated for retractile testis at prepubertal age. RESULT(S): Only 8 of 38 (21%) young adults had normal spermiograms. Five of 38 (13%) were azoospermic and 25 of 38 (66%) were oligoasthenozoospermic with ultrastructural signs of altered maturation of the sperm and a higher number of atypical forms. Of the adults with retractile testis, 2 of 7 (28.5%) were normal, 3 of 7 (43%) were oligoasthenozoospermic, and 2 of 7 (28%) were azoospermic. CONCLUSION(S): Our data support the hypothesis that prepubertal retractile testis showing signs of reduced consistency and size is a risk factor for adult infertility and requires treatment.

Adolescent↗

PubMed1994

The absent cryptorchid testis: surgical findings and their implications for diagnosis and etiology.

On surgical exploration for impalpable testes, there is often found nothing or a nubbin of tissue at the end of the spermatic vessels. This situation is commonly referred to as an absent testis. Controversy exists on how to establish correctly this diagnosis and the degree of investigation required. In addition, there is disagreement concerning whether an absent testis results from early torsion or endocrinopathic event. What is accepted is that the spermatic vessels are singularly important in establishing testis location. In this study, the pathological findings of 117 cases of absent testis diagnosed by surgical exploration at our hospital were reviewed. This diagnosis represented 10% of 1,225 patients explored for cryptorchidism from 1985 to 1991. Average patient age at operation was 26.8 months (range 5 months to 14 years). Of these children 78 (67%) presented with an impalpable left testis. At operation 3 patients (3%) underwent laparoscopy only, while all others had groin exploration with or without transperitoneal exposure to ensure identification of spermatic vessels. In 110 cases surgical specimens or nubbins were excised. Pathological study of these remnants revealed vas deferens in 89 cases (81%), epididymal tissue in 40 (36%) and small amounts of seminiferous tubules with germinal elements in 7 (6.4%). Only 26 specimens (24%) had sufficient vascular tissue present to be suggestive of spermatic vessels. A significant number showed the presence of calcification (35.5%) and hemosiderin (30%) deposits within the remnant. A subset of patients with absent testis possesses testicular tissue of presumed increased malignant potential. Therefore, surgical exploration with spermatic vessel identification and remnant removal is the gold standard for the diagnosis and treatment of the absent testis. The surgeon continues to be responsible for spermatic vessel identification, since the vessels may be recognized at pathological examination in less than 25% of the cases. Also, the common finding of calcification and hemosiderin lends weight to the torsion etiology over endocrinopathy for an absent testis.

Adolescent↗

PubMed1979

Clinical implications of gonadal venography in the management of the non-palpable undescended testis.

Selective gonadal venography was used on 28 patients with a total of 34 non-palpable undescended testes. The data obtained in this study suggest that 1) an internal spermatic vein with a pampiniform-like plexus indicates the presence of a testis, 2) a blind-ending vein on venography suggests the absence of a testis, 3) an internal spermatic vein or vas deferens may be present without a testis, 4) a testis probably cannot be present without a gonadal vein, 5) a testis may be present without a vas, 6) a blind-ending vas deferens does not necessarily indicate the absence of a testis and 7) a blind-ending vas deferens in a patient in whom a blind-ending gonadal vein is localized to the same region probably indicates the absence of a testis. Gonadal venography may localize a non-palpable undescended testis or suggest testicular agenesis. In addition, gonadal venography has aided in the selection of the operative approach and, in the future, may provide criteria under specific circumstances for determining whether an operation is necessary and, if so, the extent of surgical exploration.

Adolescent↗

PubMed2001

The nerve distribution in the testis of the cat.

The autonomous innervation of the feline testis was investigated by immunohistochemistry and a modified acetylcholinesterase technique. The nerves reach the testis mainly by two routes: (1) with testicular artery and pampiniform plexus to the cranial extremity (funicular contribution), (2) from the epididymal tail to the caudal extremity (caudal contribution). Within the tunica albuginea the funicular contribution supplies the cranial two thirds, whereas the caudal third of the tunica receives its nerves via the ligamentous connection between testis and epididymal tail. The nerve bundles accompanying the testicular artery give branches to the arterial wall and the pampiniform plexus. When reaching the cranial testicular pole the bundles separate; the majority of them pass into the centrally located mediastinum testis, another large portion enters the tunica albuginea, particularly on its epididymal side. The septula testis are innervated from both sides, that is from the mediastinum and from the tunica albuginea. In the cat, contrary to other mammals, all septula are innervated. Furthermore, nerve fibers occur regularly within the testicular lobules. Generally, the testicular nerves of the cat are unmyelinated and mainly vascular nerves, but fibers are also found within the connective tissue compartments of the testis. The vast majority of all autonomous testicular nerves are postjunctional sympathetic fibers. Terminal ramifications of cholinergic fibers are exclusively observed in the wall of medium-sized arterioles within mediastinum, septula and lobuli testis. Neuropeptide Y is the most frequent peptidergic transmitter in feline testicular vascular plexuses. The amount of calcitonin gene-related peptide-positive fibers is also remarkably high in the testis, but prefers a location within the stroma of the tunica albuginea, mediastinum and septula. In the cat, Leydig cells occur not only in intertubular locations, but also as intratunical and mediastinal Leydig cells. In all three localizations solitary nerve fibers are observed between Leydig cell groups. These fibers are generally dopamin-beta-hydroxylase- and tyrosine hydroxylase-positive, some contain calcitonin gene-related peptide and, very few, substance P.

Acetylcholinesterase↗

PubMed2000

Presenilin-1 protein specifically expressed in Leydig cells with its expression level increased during rat testis development.

Presenilin-1, mutations of which cause the early-onset of Alzheimer's disease, was shown to be abundantly expressed in the testis as well as the brain. In spite of the high expression level of this protein in the testis, no further analysis has been undertaken. We aimed to study the distribution and developmental changes in presenilin-1 protein, and to provide clues so as to elucidate the role of this protein in the rat testis. To evaluate the specificity of the anti presenilin-1 antibody, rat presenilin-1 protein was expressed in COS-7 cells and the recombinant protein was used for western blot analysis. A positive band of approximately 20 kDa corresponding to the C-terminal fragment of proteolyzed presenilin-1 protein was observed. Using testis and brain tissue samples, a 20 kDa band was detected in both tissues suggesting a similar proteolytic process, but the expression level in the testis was higher than that in the brain. The expression level increased significantly during postnatal testis development. By an immunohistochemical analysis of the rat testis, a strong signal was observed in interstitial cells and further study with cultured TM3 murine Leydig cells revealed an abundant expression of presenilin-1 in Leydig cells. Our study suggests that presenilin-1 expression in Leydig cells may play an important role in Leydig cell function and testis development.

Animals↗

PubMed1997

Characterization and cellular localization of PSG in rat testis.

In order to establish the rat testis as a model system for studying the human pregnancy-specific beta1-glycoprotein (PSG), expression and cellular distribution of PSG in rat testis were examined. Three partial PSG cDNAs, namely, rnCGM6, rnGCM7, and rnCGM8 were obtained when rat testis cDNA libraries were screened with a human placental PSG cDNA probe. Unlike the human PSGs, the rat PSGs show less nucleotide and amino acid sequence homology among family members. The rat PSGs also have multiple truncated leader sequences followed by immunoglobulin variable-like N domains while human PSGs have a single N domain. Examination of the testis, intestine, kidney, liver, lung, and muscle of male rats by reverse transcription-polymerase chain reaction (RT-PCR) with nested gene-specific primers showed that rnCGM6 was present only in the testis, while rnCGM8 was present in the testis, intestine and lung. On the other hand rnCMG7 was found in all tissues examined. Furthermore, rnCGM7 transcript was present in all somatic cells examined whereas rnCGM6 was predominantly in myoid cells and rnCMG8 in Leydig cells. These results suggest that there is cell-specificity in the expression of PSGs in the rat testis and that the rat testis is a good model for studying the biological activities of the PSGs.

Amino Acid Sequence↗

PubMed2001

The effect of alpha-tocopherol on the lipid peroxidation of mitochondria and microsomes obtained from rat liver and testis.

The effect of intraperitoneal administration of alpha-tocopherol (100 mg/kg wt/24 h) on ascorbate (0.4 mM) induced lipid peroxidation of mitochondria and microsomes isolated from rat liver and testis was studied. Special attention was paid to the changes produced on the highly polyunsaturated fatty acids C20:4 n6 and C22:6 n3 in liver and C20:4 n6 and C22:5 n6 in testis. The lipid peroxidation of liver mitochondria or microsomes produced a significant decrease of C20:4 n6 and C22:6 n3 in the control group, whereas changes in the fatty acid composition of the alpha-tocopherol treated group were not observed. The light emission was significantly higher in the control than in the alpha-tocopherol treated group. The lipid peroxidation of testis microsomes isolated from the alpha-tocopherol group produced a significant decrease of C20:4 n6 , C22:5 n6 and C22:6 n3, these changes were not observed in testis mitochondria. The light emission of both groups was similar. The treatment with alpha-tocopherol at the dose and times indicated showed a protector effect on the polyunsaturated fatty acids of liver mitochondria, microsomes and testis mitochondria, whereas those fatty acids situated in testis microsomes were not protected during non enzymatic ascorbate-Fe2+ lipid peroxidation. The protector effect observed by alpha-tocopherol treatment in the fatty acid composition of rat testis mitochondria but not in microsomes could be explained if we consider that the sum of C20:4 n6 + C22:5 n6 in testis microsomes is 2-fold than that present in mitochondria.

Animals↗

PubMed2004

Hormones: what the testis really sees.

Various barriers in the testis may prevent hormones from readily reaching the cells they are supposed to stimulate, especially the hydrophilic hormones from the pituitary. For example, LH must pass through or between the endothelial cells lining the blood vessels to reach the surface of the Leydig cells, and FSH has the additional barrier of the peritubular myoid cells before it reaches the Sertoli cells. The specialised junctions between pairs of Sertoli cells would severely restrict the passage of peptides from blood to the luminal fluid and therefore to the cells inside this barrier, such as the later spermatocytes and spermatids. There is evidence in the literature that radioactively labelled LH does not pass readily into the testis from the blood, and the concentration of native LH in the interstitial extracellular fluid surrounding the Leydig cells in rats is only about one-fifth of that in blood plasma. Furthermore, after injection with LHRH, there are large rises in LH in the blood within 15 min, at which time the Leydig cells have already responded by increasing their content of testosterone, but with no significant change in the concentration of LH in the interstitial extracellular fluid. Either the Leydig cells respond to very small changes in LH, or the testicular endothelial cells in some way mediate the response of the Leydig cells to LH, for which there is now some evidence from co-cultures of endothelial and Leydig cells. The lipophilic steroid hormones, such as testosterone, which are produced by the Leydig cells, have actions within the seminiferous tubules in the testis but also in other parts of the body. They should pass more readily through cells than the hydrophilic peptides; however, the concentration of testosterone in the fluid inside the seminiferous tubules is less than in the interstitial extracellular fluid in the testis, especially after stimulation by LH released after injection of LHRH and despite the presence inside the tubules of high concentrations of an androgen-binding protein. The concentration of testosterone in testicular venous blood does not rise to the same extent as that in the interstitial extracellular fluid, suggesting that there may also be some restriction to movement of the steroid across the endothelium. There is a very poor correlation between the concentrations of testosterone in fluids from the various compartments of the testis and in peripheral blood plasma. Determination of the testosterone concentration in the whole testis is also probably of little predictive value, because the high concentrations of lipid in the Leydig cells would tend to concentrate testosterone there, and hormones inside these cells are unlikely to have any direct effect on other cells in the testis. The best predictor of testosterone concentrations around cells in the testis is the level of testosterone in testicular venous blood, the collection of which for testosterone analysis is a reasonably simple procedure in experimental animals and should be substituted for tissue sampling. There seems to be no simple way of determining the concentrations of peptide hormones in the vicinity of the testicular cells.

Animals↗

PubMed1995

Sex-dependent expression of tenascin-C in the differentiating fetal rat testis and ovary.

The cellular mechanisms controlling sexual differentiation of fetal gonads are poorly understood. By examining the protein and mRNA expression of tenascin-C in correlation with the immunocytochemical detection of alpha-smooth muscle actin (alpha-SMA) and basement membrane heparan sulfate proteoglycan (HSPG) we demonstrate a clear-cut sex-and development-dependent expression pattern of tenascin-C in the rat testis, ovary and mesonephros. Immunocytochemistry and in situ hybridization of tenascin-C in 15-day-pc fetal testis and ovary showed protein and mRNA accumulation within the mesenchyme of the mesogonadal connection. In addition to the male and female mesonephros, some labeling could also be seen within the testicular tunica albuginea and intraovarian mesenchymal septa. In the 17-day-pc testis abundant accumulation of tenascin mRNA and protein appeared in the tunica and mediastinum testis, but not at all in the intratesticular mesenchyme. A similar pattern was still seen in the newborns where, however, a decrease in the anti-tenascin immunoreactivity of the tunica and mediastinum could be demonstrated. In contrast to the testis, expression of tenascin in 17-day-pc ovaries was widespread within the hilus and the entire intragonadal mesenchyme where it continued to accumulate also in newborns. Northern blot analysis of tenascin-C mRNAs showed one message of 8.0 kb in the 15-day-pc male and female gonads. An additional weak signal of 6.5 kb was seen in the female mesonephros. In the 18-day-pc testis, the 6.5-kb signal appeared stronger than the 8.0-kb signal. In contrast to the testis, the 6.5-kb message was weak in the developing ovary where the 8.0-kb signal had an intense peak on the day 18 pc. Further, in the ovary, mesenchymal accumulation of HSPG coincided with the spatial distribution of tenascin. In the testicular tunica and in the mesenchyme of the male and female genital ducts expression of tenascin was parallel with the differentiation of smooth muscle tissue, detected by labeling for alpha-SMA, which also indicated the tenascin-negative myoid cells of the testis. Our results indicate that tenascin expression in the fetal rat internal genitalia is involved in the differentiation of smooth muscle cells but not intratesticular myoid cells. In the ovarian mesenchyme, tenascin-C may have a specific function in the dynamic remodeling of the ovarian cords.

Actins↗

PubMed2002

The presence or absence of an impalpable testis can be predicted from clinical observations alone.

OBJECTIVE: To evaluate, in a prospective study, the accuracy of predicting the presence or absence of unilateral or bilateral impalpable testes from a clinical examination, particularly whether the contralateral descended testis (CDT) is hypertrophied. PATIENTS AND METHODS: Whether the ipsilateral scrotal appendages were palpable, and the size of the CDT, were determined before surgery in a series of patients, and compared with age-matched controls. Between 1992 and 2000, 100 impalpable testes in 86 consecutive patients (mean age at orchidopexy 45 months, range 6-223; 66% <36 months) were evaluated and treated. In addition to the presence or absence of palpable ipsilateral scrotal appendages, the size of the CDT, when present, and the intraoperative findings were recorded. Logistic regression analysis was used to model the probability of the presence or absence of the testis, as determined by the preoperative clinical findings alone. RESULTS: Of the 86 testes that were located at surgery, 50 (58%) were intracanalicular, 28 (32%) intra-abdominal and the remainder (10%) were in the superficial inguinal pouch. Of 13 patients with the 'vanishing testis syndrome', the atrophic testicle was intracanalicular in nine, in the upper scrotum in three and intra-abdominal in only one. All viable testes were successfully relocated in the scrotum, with one atrophic after surgery. The positive predictive value (PPV, with 95% confidence interval) of a testis being present when the ipsilateral appendages were palpable and the CDT was not hypertrophied was 0.93 (0.83-0.97). Conversely, the PPV of the impalpable testis being absent when the appendages were impalpable and the CDT was hypertrophied was 0.95 (0.64-0.99). CONCLUSION: When evaluating and surgically treating impalpable testes, the presence of palpable ipsilateral scrotal appendages and a CDT with no hypertrophy is associated with a 93% likelihood of discovering a testis that can be successfully relocated to the scrotum. Conversely, when the ipsilateral scrotal appendages cannot be palpated and the CDT is hypertrophied, there is a 96% probability that the impalpable testis is absent (vanishing testis syndrome). This readily available information may be valuable in preoperative counselling and planning.

Cryptorchidism↗

PubMed2002

Contralateral testicular biopsy procedure in patients with unilateral testis cancer: is it indicated?

About 5% of all patients with unilateral testis cancer harbor testicular intraepithelial neoplasia (TIN) in their contralateral testicle, which will progress into an invasive germ-cell tumor over time. Accurate diagnosis of TIN by a random surgical testis biopsy examination and effective therapy by local radiation has led to the concept of a contralateral screening biopsy procedure in all testis cancer patients. Screening and preventive treatment, however, only are indicated if (1) therapeutic outcome of the screened population is improved and (2) physiologic function of the affected organ might be maintained. Based on a critical review of the literature, some drawbacks of this policy have to be considered and the routine indication for contralateral testis biopsy procedure has to be questioned: (1) all TIN-negative patients still have to undergo meticulous follow-up evaluation for metachronous testis cancer owing to a false-negative biopsy diagnosis rate of 0.3%; (2) testis biopsy procedure is associated with a 15% to 20% complication rate, which might a negative impact on endocrine and exocrine testicular function; (3) local radiation of TIN results in irreversible infertility owing to eradication of spermatogenesis; (4) local radiation of TIN results in an impairment of endocrine Leydig cell function in 25% of patients; (5) therapeutic outcome and prognosis will not be improved in irradiated patients as compared with patients on surveillance; (6) local tumor resection for the management of metachronous testicular cancer represents an effective and viable option. The current literature does not support the strategy to perform contralateral testis biopsy procedures in all patients with unilateral testicular germ-cell tumors. Testis biopsy procedures might, however, be offered to high-risk (34%) patients for contralateral TIN with a testicular volume less than 12 mL, a history of cryptorchidism, and an age less than 30 years.

Adult↗

PubMed1996

cAMP-response element modulator tau is a positive regulator of testis angiotensin converting enzyme transcription.

Testis angiotensin-converting enzyme (ACE) is a unique form of ACE, only produced by male germ cells, and results from a testis-specific promoter found within the ACE gene. We have investigated the role of cAMP-response element modulator (CREM)tau in testis ACE transcription. In gel shift experiments, testes nuclear proteins retard an oligonucleotide containing the cAMP-response element (CRE) found at position -55 in the testis ACE promoter. Anti-CREM antibody supershifts this complex. Competitive gel shift shows that recombinant CREM tau protein and testis nuclear proteins have a similar specificity of binding to the tests ACE CRE. Functional analysis using in vitro transcription and transfection studies also demonstrate that CREM tau protein is a transcriptional activator of the testis ACE promoter. Western blot analysis identifies CREM tau protein in the protein-DNA complex formed between nuclear proteins and the testis ACE CRE motif. This analysis also identified other CREM isoforms in the gel-shifted complex, which are thought to be CREM tau 1/2, CREM alpha/beta, and S-CREM. These data indicate that CREM tau isoforms play an important role as a positive regulator in the tissue-specific expression of testis ACE.

Animals↗

PubMed1999

Testis expression of hormone-sensitive lipase is conferred by a specific promoter that contains four regions binding testicular nuclear proteins.

The testicular isoform of hormone-sensitive lipase (HSLtes) is encoded by a testis-specific exon and 9 exons common to the testis and adipocyte isoforms. In mouse, HSLtes mRNA appeared during spermiogenesis in round spermatids. Two constructs containing 1.4 and 0.5 kilobase pairs (kb) of the human HSLtes gene 5'-flanking region cloned upstream of the chloramphenicol acetyltransferase gene were microinjected into mouse oocytes. Analyses of enzyme activity in male and female transgenic mice showed that 0.5 kb of the HSLtes promoter was sufficient to direct expression only in testis. Cell transfection experiments showed that CREMtau, a testis-specific transcriptional activator, does not transactivate the HSLtes promoter. Using gel retardation assays, four testis-specific binding regions (TSBR) were identified using testis and liver nuclear extracts. The testis-specific protein binding on TSBR4 was selectively competed by a probe containing a SRY/Sox protein DNA recognition site. Sox5 and Sox6 which are expressed in post-meiotic germ cells bound TSBR4. Mutation of the AACAAAG motif in TSBR4 abolished the binding. Moreover, binding of the high mobility group domain of Sox5 induced a bend within TSBR4. Together, our results showed that 0.5 kb of the human HSLtes promoter bind Sox proteins and contain cis-acting elements essential for the testis specificity of HSL.

Animals↗

PubMedDate not supplied

Postnatal changes in the calcium binding proteins of mouse testis.

To define the changes in calcium-binding proteins (CBPs) of mouse testis during postnatal development, testicular protein extracts were analyzed by mobility shift after Ca2+ binding (MSACB) in the 2-dimensional SDS-PAGE. In the adult testis extracts, 29 kinds of protein spots showed mobility shift in the gel containing Ca2+. Among them, 2 spots showed nonspecific mobility shift and 4 spots showed Zn2+ shift. Drastic changes in expression of high-Mr CBPs (Mr > 25 kDa) were observed in testis 3 weeks after birth, suggesting that their expressions were developmentally changed. There was no obvious difference in the qualitative patterns of CBP between 4-week-old testis and adult testis. There was a large mass of CBPs (Mr = 20-25 kDa) expressed constitutively in testis throughout the postnatal development. Degree of MSACB (Mr on the diagonal line/Mr of shifted point in Ca2+ gel) of each spot was in the range of 1.1-1.7 in 12% acrylamide gel. These results suggest that changes in CBPs of testis are closely related with progression of spermatogenesis in mouse testis.

Animals↗