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New indication of testis biopsy for azoospermia: a clinical study in Japanese patients.

Because of the progress made with assisted reproductive techniques, we decided to clarify the indication for testis biopsy in Japanese azoospermic patients. A total of 88 azoospermic patients were recruited with testis histologies obtained by bilateral biopsy. Testicular histology was evaluated using Johnsen's score count. Patients with at least 1 testis containing sperm were assigned to the active spermatogenesis group. Patients whose testes had no sperm were assigned to the hypospermatogenesis group. Differences in terms of the clinical data between the 2 groups were analyzed. Clinical data consisted of past history, physical examination and hormone concentrations. The unpaired t test was generally used to examine the statistical significance of any differences between the 2 groups. 1) There were significant differences between the 2 groups in the levels of serum testosterone and luteinizing hormone. 2) There were markedly significant differences between the 2 groups in terms of testis volume and the concentration of serum follicle-stimulating hormone (FSH). 3) The smallest testis volume and the highest serum FSH value in the active spermatogenesis group were 7 ml and 32.7 mlU/ml (normal range 2.9-8.2), respectively. In conclusion, although the presence or absence of active spermatogenesis can be accurately predicted by measuring the testis volume and serum FSH, testis biopsy should be carried out in patients with a testis volume greater than 7 ml or a serum FSH less than 4 times normal when the use of assisted reproductive techniques are planned.

Adult↗

Cell-cell interactions and the regulation of testis function.

Regulatory interactions have been shown to occur between all the testicular cell types considered. The paracrine factors mediating these interactions generally influence either cellular growth or differentiation. The regulation of cellular growth is essential in the developing testis and is required for the maintenance of spermatogenesis in the adult testis. The rapid rate of germinal cell proliferation and the continuous but slowed growth of the peritubular cells and Leydig cells requires the presence of specific growth factors in the adult. Therefore, cell-cell interactions have evolved that involve growth factors such as IGF, TGF-alpha, TGF-beta and NGF. Other growth factors such as FGF or less characterized components like the seminiferous growth factor (SGF) also may be involved in the paracrine regulation of testis cell growth. An alternate cellular parameter to cell growth to consider is the regulation of cellular function and differentiation. A number of endocrine agents and locally produced paracrine factors have been shown to control and maintain testis cell function and differentiation. Cell-cell interactions mediated by factors such as androgens, POMC peptides, and PModS are all primarily directed at the regulation of cellular differentiation. Therefore, the agents which mediate cell-cell interactions in the testis can generally be categorized into factors that regulate cell growth or those which influence cellular differentiation. The specific cell-cell interactions identified will likely be the first of a large number of cellular interactions yet to be investigated. Although a number of potentially important cell-cell interactions have been identified, future research will require the elucidation of the in vivo physiological significance of these interactions. The existence of different cell types and potential cell-cell interactions in a tissue implies that the actions of an endocrine agent on a tissue will not simply involve a single hormone and single cell. The endocrine regulation of testis function will have effects on cell-cell interactions and be affected by local cell-cell interactions. The ability of LH to influence Leydig cell androgen production promotes a cascade of interactions mediated through several cell types to maintain the process of spermatogenesis. FSH actions on Sertoli cells also promote cell-cell interactions that influence germinal cell development, peritubular myoid cell differentiation and Leydig cell function. Therefore, elucidation of the endocrine regulation of testis function requires an understanding of the local cell-cell interactions in the testis.

Animals↗

Cancer-testis genes are coordinately expressed and are markers of poor outcome in non-small cell lung cancer.

PURPOSE: Cancer-testis genes mapping to the X chromosome have common expression patterns and show similar responses to modulators of epigenetic mechanisms. We asked whether cancer-testis gene expression occurred coordinately, and whether it correlated with variables of disease and clinical outcome of non-small cell lung cancer (NSCLC). EXPERIMENTAL DESIGN: Tumors from 523 NSCLC patients undergoing surgery were evaluated for the expression of nine cancer-testis genes (NY-ESO-1, LAGE-1, MAGE-A1, MAGE-A3, MAGE-A4, MAGE-A10, CT7/MAGE-C1, SSX2, and SSX4) by semiquantitative PCR. Clinical data available for 447 patients were used to correlate cancer-testis expression to variables of disease and clinical outcome. RESULTS: At least one cancer-testis gene was expressed by 90% of squamous carcinoma, 62% of bronchioloalveolar cancer, and 67% of adenocarcinoma samples. Statistically significant coexpression was observed for 34 of the 36 possible cancer-testis combinations. Cancer-testis gene expression, either cumulatively or individually, showed significant associations with male sex, smoking history, advanced tumor, nodal and pathologic stages, pleural invasion, and the absence of ground glass opacity. Cox regression analysis revealed the expression of NY-ESO-1 and MAGE-A3 as markers of poor prognosis, independent of confounding variables for adenocarcinoma of the lung. CONCLUSIONS: Cancer-testis genes are coordinately expressed in NSCLC, and their expression is associated with advanced disease and poor outcome.

Antigens, Neoplasm↗

Tissue guanosine-3',5'-cyclic monophosphate levels and soluble guanylate cyclase activity: a positive correlation during unilateral cryptorchidism in the rat testis.

The relationship between the subcellular distribution of guanylate cyclase and tissue guanosine-3',5'-cyclic monophosphate (cGMP) levels was investigated in rat testes after surgically induced unilateral cryptorchidism. Placement of one of a testis pair in the abdominal cavity results in loss of testicular weight and function in the abdominal testis whereas the remaining scrotal testis appears to be functionally normal. Within 5 days after surgery, tissue cGMP levels were increased by twofold in the abdominal testis. A fourfold elevation was noted from 10 to 30 days after surgery. Whereas the homogenate guanylate cyclase activity was only slightly elevated 10 and 20 days postoperatively, a 200% increase in the soluble guanylate cyclase activity was seen at 5 days. Between 10 and 30 days, the rise in activity was >250% (P < 0.01). An increase in soluble guanylate cyclase activity was noted when the data were expressed as per milligram protein, per milligram DNA or per whole testis. Conversely, particulate guanylate cyclase activity was reduced by 40% in the cryptorchid testis. Kinetic analysis of the soluble enzyme prepared from abdominal and scrotal testes yielded linear Line-weaver-Burke plots for both enzyme preparations with an identical K(m) for guanosine triphosphate, but a three-fold higher maximal velocity for the abdominal enzyme. When the soluble guanylate cyclases from both testes were mixed and assayed together, the activities were additive rather than exhibiting synergism or inhibition. These experiments indicate that the altered V(max) is not due to a transferable activator or inhibitor.An immunocytochemical technique was used to assess the cell type in which the alterations in cGMP metabolism occurred. Comparison of the scrotal and abdominal testes revealed that the abdominal testis exhibited enhanced cGMP immunofluorescence within the cells lining the inner aspect of the seminiferous tubule as well as tubular elements and interstitial cells. Thus, it is inferred that the correlated changes in soluble guanylate cyclase activity and cGMP levels occur in several of the cell types that remain viable within the cryptorchid testis.

Animals↗

Bilateral cystic rete testis in an alpaca (Lama pacos).

A 9-year-old intact male alpaca (Lama pacos) was examined because of marked enlargement of the left scrotum. Ultrasound examination revealed a thin-walled anechoic structure in the area of the left testis. Aspirated fluid contained spermatozoa, many of which had abnormal morphology. Castration was performed and the left testis was markedly enlarged with a clear fluid-filled cyst. The cyst was lined by a single layer of squamous to cuboidal epithelial cells consistent with those originating from rete testis. The right testis was of a comparable size and shape to that of normal alpaca testis, but the rete testis was mildly to moderately dilated. Additional findings included chronic inflammation of the right testis and epididymis and epididymal fibrosis with ductal hyperplasia on the left. The diagnosis was bilateral cystic rete testis, most likely secondary to chronic inflammation.

Animals↗

Phospholipid-sensitive Ca2+-dependent protein kinase system in testis: localization and endogenous substrates.

The presence and localization in pig and rat testes of phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) and its substrates were investigated. PL-Ca-PK activity was found in the testis total particulate fraction and epididymal fluid, but little in the testis total particulate fraction and epididymal fluid, but little in the testis cytosol and matured spermatozoa obtained from the epididymis. Similarly, at least three endogenous substrates (83,000, 33,000, and 26,000) and five substrates (greater than 100,000, 83,000, 60,000, 43,000, and 19,000) for PL-Ca-PK were detected in the testis total particulate fraction and the epididymal fluid, respectively, but little or no substrates were observed in the testis cytosol and matured spermatozoa. The three substrates detected in the total particulate fraction were also observed in the testis nuclear fraction. In rat testis, PL-Ca-PK activity was detected in the total particulate fraction of germ cells. The results suggested that PL-Ca-PK system might be important in membrane- or subcellular organellar-associated functions in testis.

Animals↗

Corticosteroid 11 beta-dehydrogenase in rat testis.

Corticosteroid 11 beta-dehydrogenase, the enzyme that catalyzes the oxidation of the biologically active steroid cortisol to its inactive metabolite cortisone, is present in testis. Since excess cortisol in men and other mammals and excess corticosterone in rodents cause physiological abnormalities including abnormal testicular function, it was pertinent to study the cellular distribution of 11 beta-dehydrogenase in the testis. Purified antiserum directed against homogeneous rat 11 beta-dehydrogenase was used to localize the enzyme in the developing rat testis. With immunofluorescence, the enzyme was not detectable in fetal testis or in the testis of young male rats until the 26th day of development. A few interstitial cells were stained in the testis of 26-day-old animals. In the testis of 31-day-old rats many cells in the interstitium were positive. In adult animals the entire interstitial region displayed bright fluorescence. Depleting animals of germ cells did not abolish the fluorescence. The appearance of this enzyme correlates temporally with the postnatal increase in Leydig cell number and the developmental rise in serum testosterone. We suggest that 11 beta-dehydrogenase of Leydig cells protects the testis from the deleterious effects of cortisol.

11-beta-Hydroxysteroid Dehydrogenases↗

Identification of GABABR2 in rat testis and sperm.

GABA is capable of mimicking and potentiating the action of progesterone in initiating the acrosome reaction (AR) of mammalian sperm. The GABA-initiated AR is mediated by GABA(A)R; whereas GABA(B)R1 protein found in rat testis and sperm tends to modify this process. Moreover, the occurrence of GABA(B)R2, a subunit essential for the formation of a functionally active GABA (B)R, in rat testis and sperm has not been established. In the present study, rat testis and sperm were analyzed for the presence of GABA(B)R2 transcript and protein by RT-PCR, Northern blot, Western blot and an indirect immunofluorescence technique. Northern blot shows that the transcript of testis GABA(B)R2 is shorter (~3.0 Kb) than that of the brain (~5.6 Kb). The full length testis GABA(B)R2 cDNA was prepared by RACE-PCR and found to be shorter by 2.2 Kb in the segment at the extreme terminus of 3'UTR of rat brain GABA(B)R2 but, the sequences corresponding to the open reading frame and 5'-UTR of rat testis GABA(B)R2 were found to be identical to those of rat brain. GABA(B)R2 protein isolated from rat epididymal sperm was slighter larger than those of rat testis and brain. It was principally localized in the acrosome region of the head of rat sperm by an indirect immunofluorescence technique. The present results establish that GABA(B)R2 protein is produced in rat testis and sperm and may play a role in GABA triggering of AR.

Acrosome Reaction↗

Expression of Hsp70-2 in unilateral cryptorchid testis of rhesus monkey during germ cell apoptosis.

We investigated the possible role of Hsp70-2 in germ cell apoptosis induced by heat stress in monkey unilateral cryptorchid testis. The study focused on in situ analysis of the testicular cell DNA fragmentation and on the possible relationship between Hsp70-2 expression and germ cell apoptosis. The TUNEL result showed that most of the germ cells were labeled in the cryptorchid testis on d 5 after induction of cryptorchidism; that with most of the apoptotic germ cells depleted, only a few germ cells were labeled on d 10; and that almost no apoptotic signal was observed in the cryptorchid testis on d 15 and thereafter. This indicates that the increasing germ cell degeneration in cryptorchid testis may take the form of apoptosis. Using in situ hybridization, immunohistochemistry, and Northern blot, we examined the changes of Hsp70-2 expression in the monkey cryptorchid testis. The level of Hsp70-2 mRNA decreased slightly, while the expression of HSP70-2 protein was almost unchanged at the early stage of germ cell apoptosis in the cryptorchid testis on d 5 and dropped dramatically along with the loss of apoptotic germ cells in the cryptorchid testis on d 10 after operation. It is therefore suggested that Hsp70-2 might not take part in inhibiting the apoptosis of germ cells at the early stage during operation-induced cryptorchid testis, and that Hsp70-2 gene does not belong to the immediate early related gene responsible for germ cell apoptosis induced by heat stress.

Animals↗

Changes of diurnal rhythm and levels of total and free testosterone secretion from pre to late puberty in boys: testis size of 3 ml is a transition stage to puberty.

OBJECTIVE: To establish levels for comparison for 24-h total and free serum testosterone in prepubertal boys and throughout pubertal development. DESIGN: The study subjects were 55 healthy boys, aged 5.0-18.6 years, who underwent serial sampling one or more times during their pubertal development. METHODS: Testicular volumes were determined by orchidometer. Serum testosterone was measured by a modified RIA (detection limit, 0.03 nmol/l). Free testosterone was calculated (calc-FT) using a formula derived from the law of mass action. RESULTS: Significant increases in testosterone and calc-FT concentrations in boys were found between testis volumes of 1 ml to 2 ml, 2 ml to 3 ml, 6 ml to 8 ml, and 10 ml to 15 ml. No differences were found between testis volumes of 3, 4, 5 and 6 ml neither were there differences between 8 and 10 ml, or between 15, 20 and 25 ml. Boys who had reached their final height had higher calc-FT values than boys who had the same pubertal development but had not reached their final height. Based on the results, puberty was classified into six stages: pre1 (testis, 1 ml), pre2 (testis, 2 ml), early (testis, 3-6 ml), mid (testis, 8-12 ml), late1 (testis,15-25 ml, not reached final height) and late2 (testis, 15-25 ml, reached final height). Serum testosterone was secreted with a diurnal variation in prepuberty and during puberty. The increase of testosterone in the morning hours started earlier in pubertal than in pre-pubertal boys. The most pronounced diurnal rhythm was found in early and in mid puberty. CONCLUSION: Using a sensitive method, and a pubertal reclassification, we have established levels for comparison of testosterone and calc-FT in prepubertal and pubertal boys. The existence of data for comparison forms the basis for future studies on pubertal disorders.

Adolescent↗

Effects of neonatal administration of the reversible goitrogen propylthiouracil on the testis interstitium in adult rats.

The effects of neonatal treatment with the reversible goitrogen, 6-n-propyl-2-thiouracil (PTU) on the volumes of testicular interstitial components, the number and average volume of Leydig cells, and the steroid secretory capacity of testis and Leydig cells of rats at 135 days of age were investigated. Rat pups were hypothyroid from birth to 25 days of age following the addition of 0.1% (w/v) PTU to the drinking water of the mother. Treatment was stopped at 25 days and the pups subsequently returned to a euthyroid state. Control rat pups were raised without adding PTU to drinking water of the mother. On day 135, one testis from each rat (n = 5 per group) was fixed by whole body perfusion for microscopy and stereology, and the ipsilateral testis was used for steroid secretion analysis using an in vitro testis perfusion system. Average testis volume was 84% greater in PTU-treated rats than in controls. This increase resulted from increases in both the seminiferous tubule (86%) and the interstitial (80%) volumes. Moreover, absolute volumes of all testis components in PTU-treated rats were significantly (P < 0.05) greater than those of controls; the highest volume increase was observed in the lymphatic space (147%). The number of Leydig cells per testis was nearly doubled, but the average volume of a Leydig cell was decreased by 25% in PTU-treated rats compared with controls. Steroid secretion per testis was not significantly different between control and PTU-treated rats; however, steroid secretion per Leydig cell was significantly lower in PTU-treated rats than in controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Androgens↗

Developmental and hormonal regulation of type II DNA topoisomerase in rat testis.

Type II DNA topoisomerase (topo II) is required for diverse biological functions including DNA replication, maintenance of genome stability, chromosome segregation and chromosome condensation. While the identity of topo II in rodent testis has been established, the regulation of topo II expression during the development of the postnatal testis and gametogenesis is unclear. Here, we report that rat testis topo II is developmentally and hormonally regulated. Topo IIalpha mRNA levels peaked prior to the onset of puberty, declined sharply thereafter and stabilized in adult testis. In contrast, the topo II enzyme content was lower in prepubertal testis but increased after the onset of puberty. Topo II was expressed in a cell-specific manner within germ cells, being detected only in pachytene spermatocytes. While testosterone markedly increased topo IIalpha mRNA levels in prepubertal testis, continued treatment failed to enhance topo IIalpha mRNA above postpubertal control levels. The extent of topo II activity remained steady regardless of the testosterone-induced increase in topo IIalpha mRNA levels. Inhibition of testosterone function in postpubertal animals by ethanedimethane sulphonate (EDS) and flutamide resulted in a significant decrease in topo IIalpha gene expression and topo II activity. The administration of exogenous testosterone (T) to EDS- and flutamide-treated rats restored topo IIalpha mRNA levels and topo II activity similar to the levels seen in the testis of age-matched control animals. Histochemical analyses of testes indicated that the effect of T on spermatogenesis was separable from its effect on topo IIalpha expression. Our results reveal that testosterone acts as a positive regulator of topo IIalpha gene expression and is required for the maintenance of topo IIalpha expression during the development of the postnatal testis and spermatogenesis.

Animals↗

Available FSH receptors in adult rat testis in vivo.

It has been suggested that the tight junctions formed between Sertoli cells during the peripubertal period constitute a barrier to circulating FSH in the adult testis, limiting its access to the lumen of the seminiferous tubules. There are also reports of FSH receptor-binding inhibitors. These observations prompted us to study the extent of FSH receptor availability in vivo in the adult rat. Experimental rats were given an intracardiac injection of rat FSH (rFSH), and the occupied receptor was measured by radioimmunoassay of acid-released rFSH from the testis. In the saline-injected control animals, there were 247 fmol occupied FSH receptors/g testis, and as much as 1788 fmol of the unoccupied high-affinity receptors/g testis, as measured by in-vitro binding studies. After intracardiac injection of increasing amounts of rFSH (up to 606 pmol), receptor occupancy increased to a maximum plateau of only 448 fmol/g testis. In contrast, when rFSH was given by intratesticular injection in order to achieve pharmacological doses in the testis, the maximum binding was 662 fmol/g testis. Scatchard analysis of the in-vivo data revealed, however, that the maximum concentration of the high-affinity receptor was 452 fmol/g testis, a value concordant with the highest in-vivo binding observed in animals given intracardiac rFSH (448 fmol/g). A single injection of the hormone did not induce down-regulation of FSH receptors, regardless of the dose, whereas multiple injections of menotrophin were effective, at least to some extent. Despite the receptor loss, the immediate receptor availability was maintained, suggesting the presence of a receptor pool.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Induction of oxidative stress by organic hydroperoxides in testis and epididymal sperm of rats in vivo.

The present study describes the extent and pattern of oxidative stress induction in testis and epididymal sperm of rats following in vivo exposure to repeated sublethal doses of 2 model pro-oxidants, namely, t-butyl hydroperoxide (tbHP) and cumene hydroperoxide (cHP). Single sublethal (1/40, 1/20, and 1/10 LD(50)) doses of hydroperoxides (HP) administered intraperitoneally to male rats (CFT-Wistar strain) failed to induce any significant increase in malondialdehyde or reactive oxygen species (ROS) levels in testis or epididymal sperm. However, repeated doses for 1 or 2 weeks induced a marked dose-related enhancement of lipid peroxidation (LPO) and ROS levels in both testis and epididymal sperm. Further evidence, such as significant perturbations in both enzymic and nonenzymic antioxidants and enhanced levels of protein carbonyls in testis, suggested induction of oxidative stress. In testis, moderate depletion in reduced glutathione levels and marked diminution in ascorbic acid and alpha-tocopherol content were accompanied by increased activities of various antioxidant enzymes, namely glutathione peroxidase, glutathione-S-transferase, and catalase, in both the HP treatments. Furthermore, significant alterations in the specific activities of testicular enzymes such as LDH-X, G-6-PDH, and SDH indicated altered testicular physiology. Both HP at higher doses induced significant DNA damage (determined by fluorimetric analysis of DNA unwinding assay) in testis and epididymal sperm. Increased total iron levels in testis of HP-treated rats are indicative of the possible involvement of iron-mediated free radical reactions in this model. These findings provide an account of early oxidative damage in testis and epididymal sperm following short-term exposure to HP in vivo, and this model is being further exploited for understanding the consequences of chronic oxidative stress-mediated alterations for the physiology of male reproductive system and its implications for fertility.

Animals↗

Testicular development and endocrine characteristics of boars selected for either high or low testis size.

Thirty-six Landrace x Large White cross boars were selected from litters with either high or low estimated breeding values for 150-d paired testis weight. Blood samples were taken via jugular venipuncture at eight ages (42, 56, 70, 84, 98, 112, 126 and 140 d). At each sampling age, nine blood samples were taken at 30-min intervals. Luteinizing hormone (LH) was determined on the individual serum samples. Serum samples from each boar at each age were pooled and concentrations of follicle-stimulating hormone (FSH), estradiol-17 beta (E2) and testosterone (T) were determined. Paired testis width, testis length and body weight were measured at 98, 112, 126 and 140 d of age. Backfat probe, weights of excised testes and histological data on testes were obtained at 140 d of age. Boars with high testis weight (HTW) were heavier (P less than .05), had higher adjusted backfat probes (P less than .01) and had consistently larger in situ testis measurements (P less than .01) than did low testis weight (LTW) boars. Boars with HTW had heavier (P less than .01) testes and epididymides at 140 d of age. They also had a higher percentage of seminiferous tubules in which spermatogenesis was present (P less than .05), a larger percentage of tubules with a lumen (P less than .05) and tubules had a larger mean diameter (P less than .01) than did those of boar with LTW. Adjustment of in situ testis measurements and excised testis weights for body weight reduced line differences by less than 20%. A rise in LH concentrations occurred at approximately 100 d of age. Boars with HTW had higher (P less than .05) and more variable (P less than .01) LH concentrations than did boars with LTW. Boars with HTW also had higher maximum concentrations of LH during the pubertal rise (P less than .01) and these concentrations tended to reach maximum levels at younger ages. Concentrations of T increased in a fashion that was nearly linear with age (P less than .01) and tended to be higher for the boars with HTW (P less than .10). Concentrations of E2 changed little from 42 to 84 d of age but increased steadily thereafter. Boars with HTW had a more rapid increase in E2 concentrations than did boars with LTW (P less than .05).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Acute and sub-acute effects of 2-butenoic acid-3-(diethoxy phosphinothioyl) methyl ester (RPR-II) on testis of albino rat.

Acute and sub-acute toxic effects of a novel phosphorothionate coded as RPR-II on testis of albino rats were studied. In acute study rats received a single dose of 12.3 mg/kg of RPR-II and sacrificed after 24 hr. For sub-acute study 0.58 mg/kg/day was administered orally to rats for 10 and 21 days. Acute exposure of rats to RPR-II brought no change either in the gonadosomatic index (GSI) or in the structure of testis or in the serum levels of testosterone. Testis glutathione (GSH) level and glutathione S-transferase (GST) activity was significantly decreased whereas, acid phosphatase (AcP) levels increased significantly at 24 hr post-treatment. On 7th day (withdrawal period) after the cessation of the treatment the GSH, GST, AcP, and AkP levels reached to near control. The sub-acute study revealed a significant decrease in GSI on 10th and 21st day of the treatment. In contrast, a time-dependent and significant increased in GSH level and GST activity was observed on 100th and 21st day of post-treatment, except GSH level on 10th day, which was declined. Due to RPR-II treatment the testis AcP and alkaline phosphatase (AkP) levels were significant at both 10th and 21st day of medication but AcP levels were increased whereas AkP levels decreased. The histopathological studies on day 10th showed considerable loss of spermatozoids in testis and at 21st day complete derangement of cellular organization was observed. Testosterone levels decreased significantly on 10th day and remained significantly low at 21st day. However, withdrawal studies showed a recovery in testis of rat treated with RPR-II. GST, GSH, GSI, AcP and AkP values recovered, testosterone levels were also well recovered but recovery in testis structure remained at a low profile. The present study suggests that RPR-II may cause testicular toxicity in rats affecting the normal functioning of testis and it also gave some new information in withdrawal studies.

Alkaline Phosphatase↗

Expression of a novel bHLH-Zip gene in human testis.

AIM: To identify specifically expressed genes in the adult and fetal testes. METHODS: A human testis cDNA microarray was established. Then the mRNA of adult and fetal testis was purified and probes were prepared by a reverse transcription reaction with the testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes. The nucleic sequences of differentially expressed genes were determined and homologies were searched in the databases of the GenBank. RESULTS: When hybridized with adult or fetal testis probes, the positive clones were 96.8 % and 95.4 %, respectively. Among these genes, one was a new testis-specific gene, which was named TSP1. TSP1 was highly expressed in human adult testis. The cDNA of TSP1 was 1,484 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AF333098). TSP1 was also determined as Interim Gen Symbol (Unigene, No. Hs.98266). Protein analysis showed that TSP1 contained two functional domains: an N-terminal basic helix-loop-helix (bHLH) and a C-terminal leucine zipper (Zip). Homologous analysis showed that the 430 amino acid sequences deduced from the 1293 bp open reading frame (ORF) had a homology with the human gene FLJ2509 (AK098575). TSP1 had also a sequence homology with Spz 1 protein of mouse. Expression profiles showed that TSP1 was specifically and strongly expressed in the testis. CONCLUSION: TSP1 is a gene highly expressed in adult testis. It may play an important role in spermatogenesis in the humans.

Adult↗

Antioxidant enzyme activity in the maturing rat testis.

Developmental profiles of the activity of the antioxidant enzymes superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px), glutathione transferase (GSH-Tr), and hexose monophosphate shunt (HMS) were measured in the rat testis and liver. The level of SOD in the testis was high at the age of 6 to 10 days, after which it dropped to approximately one third of that level by 20 days of age, and remained there up to 8 months of age. In the liver, SOD activity steadily increased from the neonatal to adult stage of life, reaching the same level as detected in the testis. The testicular activity of catalase was only 2% to 7% of that found in liver at all ages. It increased in both organs up to 6 weeks of age, whereafter the hepatic activity gradually decreased and no further changes were seen in the testis. The GSH-Px activity was low in the testis and declined slightly with age, whereas activity in the liver increased four-fold between birth and adulthood. The activity of GSH-Tr was similar in both organs studied: it increased after birth, showing a maximum in the liver at 1.5 months (ten-fold increase) and in the testis at 5 months of age (four-fold increase). The HMS activity was two to three times higher in the liver than in the testis, and decreased slightly with age in both organs. Thus, the basal levels and developmental profiles of antioxidant enzymes in the testis differ greatly from those in the liver.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗