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Microtubules of the mouse testis exhibit differential sensitivity to the microtubule disruptors Carbendazim and colchicine.

The testicular toxicant benomyl and its metabolite, carbendazim cause reproductive damage to the rat, an early sign of which is sloughing of germ cells with associated Sertoli cell fragments. However, the sensitivity of other mammalian species to these benzimidazole compounds is not clear. In this study, the effects of carbendazim and colchicine, a known microtubule disruptor, on the mouse seminiferous epithelium were characterized, and the amount of carbendazim reaching the mouse testis was measured. Testes were assessed for histological effects 3 h and 6 h after administration of carbendazim (2000 mg/kg, ip), and 6 h after intratesticular administration of either a low or high dose (5.3 or 117.6 micro g/g testis) of colchicine. Carbendazim caused no signs of histological damage to the mouse testis, and the microtubule cytoskeleton was intact and identical to controls based on immunostaining with tyrosinated alpha tubulin and beta tubulin antibodies. Similarly, the seminiferous epithelium of mouse testis was undamaged and the microtubule cytoskeleton was intact after a low dose of colchicine, while a comparable dose of colchicine injected into rat testis caused marked toxicity. However, mouse testes did show microtubule disruption and severe germ cell sloughing after administration of a high dose of colchicine. The amount of carbendazim measured in mouse testis was 375 nmol/g testis, which is higher than the value measured in rat testis after a toxic dose of carbendazim. Therefore, carbendazim reaches the mouse testis at or above levels measured in the rat, yet the mouse is apparently insensitive to this microtubule disrupting agent.

Animals↗

Testis morphometry, seminiferous epithelium cycle length, and daily sperm production in domestic cats (Felis catus).

There is very little information regarding the testis structure and function in domestic cats, mainly data related to the cycle of seminiferous epithelium and sperm production. The testis weight in cats investigated in the present study was 1.2 g. Compared with most mammalian species investigated, the value of 0.08% found for testes mass related to the body mass (gonadosomatic index) in cats is very low. The tunica albuginea volume density (%) in these animals was relatively high and comprised about 19% of the testis. Seminiferous tubule and Leydig cell volume density (%) in cats were approximately 90% and 6%, respectively. The mean tubular diameter was 220 microm, and 23 m of seminiferous tubule were found per testis and per gram of testis. The frequencies of the eight stages of the cycle, characterized according to the tubular morphology system, were as follows: stage 1, 24.9%; stage 2, 12.9%; stage 3, 7.7%; stage 4, 17.6%; stage 5, 7.2%; stage 6, 11.9%; stage 7, 6.8%; and stage 8, 11 %. The premeiotic and postmeiotic stage frequency was 46% and 37%, respectively. The duration of each cycle of seminiferous epithelium was 10.4 days and the total duration of spermatogenesis based on 4.5 cycles was 46.8 days. The number of round spermatids for each pachytene primary spermatocytes (meiotic index) was 2.8, meaning that significant cell loss (30%) occurred during the two meiotic divisions. The total number of germ cells and the number of round spermatids per each Sertoli cell nucleolus at stage 1 of the cycle were 9.8 and 5.1, respectively. The Leydig cell volume was approximately 2000 microm3 and the nucleus volume 260 microm3. Both Leydig and Sertoli cell numbers per gram of testis in cats were approximately 30 million. The daily sperm production per gram of testis in cats (efficiency of spermatogenesis) was approximately 16 million. To our knowledge, this is the first investigation to perform a more detailed and comprehensive study of the testis structure and function in domestic cats. Also, this is the first report in the literature showing Sertoli and Leydig cell number per gram of testis and the daily sperm production in any kind of feline species. In this regard, besides providing a background for comparative studies with other fields, the data obtained in the present work might be useful in future studies in which the domestic cat could be utilized as an appropriate receptor model for preservation of genetic stock from rare or endangered wild felines using the germ cell transplantation technique.

Animals↗

Ischemia-reperfusion of the murine testis stimulates the expression of proinflammatory cytokines and activation of c-jun N-terminal kinase in a pathway to E-selectin expression.

Ischemia-reperfusion (IR) of the testis results in germ cell-specific apoptosis and can lead to aspermatogenesis. Germ cell-specific apoptosis after IR of the testis has been shown to be correlated with and dependent on neutrophil recruitment to the testis after IR. Studies that used E-selectin-deficient mice have demonstrated that E-selectin expression is critical for neutrophil recruitment to subtunical venules in the testis after IR and for the resultant germ cell-specific apoptosis. The present study investigates the in vivo signaling pathway that exists after IR that leads to neutrophil recruitment in the murine testis. Mice were subjected to a 2-h period of testicular ischemia followed by reperfusion. Results demonstrate that the proinflammatory cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta), are stimulated after IR as is the phosphorylation of c-jun N-terminal kinase (JNK). The downstream transcription factors of JNK, ATF-2 and c-jun are also phosphorylated at specific times after IR of the testis. Activation of the JNK stress-related kinase pathway is correlated with an increase in E-selectin expression and neutrophil recruitment to the testis after IR. Intratesticular injection of IL-1beta also caused JNK phosphorylation and neutrophil recruitment to the testis. These results suggest that testicular IR injury stimulates IL-1beta expression, which leads to activation of the JNK signaling pathway and ultimately E-selectin expression and neutrophil recruitment to the testis. This provides the first evidence of a cytokine/stress-related kinase signaling pathway to E-selectin expression in vivo.

Activating Transcription Factor 2↗

Grafting period and donor age affect the potential for spermatogenesis in bovine ectopic testis xenografts.

Bovine testis tissue xenografts contain elongating spermatids 6 mo after grafting. The percentage of seminiferous tubule cross sections with elongating spermatids at the time of graft removal varies depending on donor age and rarely exceeds 10%. These data indicate significant changes are occurring to bovine testicular cells during the first weeks of life. The objective of this research was to xenograft testis tissue from multiple ages of bull calves for 24 or 36 wk in order to gain a better understanding of early bovine testis development. Testis tissue from 1-, 2-, 4-, and 8-wk-old calves was grafted onto the backs of castrated immunodeficient mice. Testis tissue from all donor ages grew, differentiated, and produced testosterone and elongating spermatids. Testis tissue grafts from 1- and 8-wk-old calves had elongating spermatids in greater than 5.5% of seminiferous tubule cross sections at the time of graft removal regardless of grafting period. Four-week-old donor tissue never had more than 5.2% of seminiferous tubule cross sections with elongating spermatids. Extending the grafting period from 24 to 36 wk resulted in an increase in the percentage of seminiferous tubule cross sections with elongating spermatids from 2% to 10% in 2-wk donor tissue. These data demonstrate that both donor age and grafting period may be important factors regulating the maturation of bovine testis xenografts, indicating that intrinsic differences exist within testis tissue at these donor ages. These data provide the framework for further study of bovine spermatogenesis using ectopic testis xenografting.

Age Factors↗

Testis histopathology in boys with cryptorchidism correlates with future fertility potential.

PURPOSE: We studied the ability of testis biopsy to predict eventual fertility potential. MATERIALS AND METHODS: A total of 25 patients with unilateral undescended testis and 11 with bilateral undescended testes who previously underwent orchiopexy at a median age of 9 years were followed until after age 18 years. Semen analysis, testicular volume, follicle-stimulating hormone, luteinizing hormone, testosterone and inhibin B were compared to previously obtained biopsies of each testis. Biopsies were graded as mild, moderate or severe histology based on published data for normal and undescended testis histology. RESULTS: Patients with unilateral undescended testis and moderate histology were more likely to have a sperm density of greater than 20 million per cc than those with severe histology (p = 0.006), although no difference in hormonal parameters was noted. Patients with bilateral undescended testes and moderate histology were more likely to have normal follicle-stimulating hormone and inhibin B than those with severe histology (p = 0.05 and 0.002, respectively), although no difference in semen analysis was observed. Overall 7 of the 9 patients with bilateral undescended testes had less than 20 million sperm per cc. CONCLUSIONS: Testis biopsy provides useful prognostic information about semen analysis in patients with unilateral undescended testis. In the 9 patients with bilateral undescended testes testis biopsy divided them into normal and abnormal groups in terms of hormonal parameters. Testis biopsy was not able to provide additional prognostic information on semen analysis in patients with bilateral undescended testes since they were uniformly poor.

Adolescent↗

Is an empty left hemiscrotum and hypertrophied right descended testis predictive of perinatal torsion?

PURPOSE: We determine whether the finding of a nonpalpable left testis and hypertrophied(2 cc or greater) right testis is predictive of an atrophic (perinatal torsion) left testis. MATERIALS AND METHODS: Prospectively, all boys with a nonpalpable left testis and hypertrophied right testis seen between May 2000 and May 2002 were included in the study. Testicular measurement was performed preoperatively with an orchidometer. In 19 of 22 boys size was also confirmed intraoperatively. Surgical exploration was done initially through a scrotal incision. Diagnostic laparoscopy was performed in boys in whom intrascrotal tissue that was consistent with a "nubbin" was not found. All tissue removed was submitted for histological evaluation. RESULTS: In 19 of 22 boys tissue was found in the left hemiscrotum that was clinically consistent with a scrotal nubbin, and histological confirmation was absolute in 18. In 1 patient a hollow oval mass attached to a cord extending to the external inguinal ring was found without the other classic histological features of torsion. In 3 cases scrotal exploration was negative and diagnostic laparoscopy was performed. Of this group the pathognomonic findings of a closed internal ring with hypoplastic vas and vessels were noted in 2 cases, and normal vas and vessels were seen to exit an open internal inguinal ring in 1. Inguinal exploration demonstrated an enlarged intracanalicular testis measuring 3 cc in volume, comparable in size to the descended right testicle. CONCLUSIONS: The combination of a nonpalpable left testis and an enlarged right testis is highly predictive of perinatal testicular torsion. When both criteria were met 20 of 22 (91%) consecutive patients had histological or laparoscopically confirmed perinatal torsion and 1 had only clinical features. This finding supports the concept of scrotal exploration as the initial procedure in the child who has an empty left hemiscrotum and hypertrophied descended right testis. Laparoscopy should be reserved for boys in whom a distinct remnant is not found on scrotal exploration.

Atrophy↗

Pagetoid spread into the rete testis by testicular tumours.

Germ cell neoplasia of the testis can spread into the rete testis in a pagetoid manner, but this is less often recognized than vascular invasion or direct spread into the tunica albuginea. The rete testis was studied in 71 orchidectomy specimens: 47 with an untreated germ cell tumour, 18 with germ cell neoplasia after treatment with chemotherapy, and six with other tumours. Pagetoid spread into the rete testis was seen in 14 of the 47 cases of germ cell tumour. The cytology of the infiltrating cells was that of intratubular germ cell neoplasia. Hyperplasia of the rete testis was seen in six cases. In three of the four testes with untreated germ cell neoplasia and hyperplasia of the rete, neoplastic cells were seen in the hyperplastic rete epithelium. The two testes with extensive pagetoid involvement of the rete testis by intratubular germ cell neoplasia both had associated hyperplasia of the rete testis. Two of the 18 testes with germ cell neoplasia after chemotherapy had focal hyperplasia. Pagetoid involvement of the rete testis by intratubular germ cell neoplasia is common and may be a cause of hyperplasia of the rete testis.

Humans↗

The effect of experimental cryptorchidism on the phosphorus NMR spectrum of the rat testis.

Magnetic resonance (MR) spectroscopy of the cryptorchid rat testis was used to test whether changes in the MR spectra of the rat testis might be a more sensitive indicator of changes in the metabolic status of germ cells in the testis rather than simply the cell types present. Testes of adult Wistar rats before and during 42 days of experimental cryptorchidism were investigated by in-vivo 31P MR spectroscopy. Results were compared to MR studies of the synchronized developing testis. The testicular phosphomonoester/ATP (PM/ATP) ratio was dependent only on the cell types present, and showed the same characteristics for each cell type present in the degenerating testis as in the developing testis. The testicular phosphodiester/ATP (PD/ATP) ratio decreased rapidly when the number of round and elongated spermatids was reduced. Similar effects, although less pronounced, were seen in the developing testis. The pH decreased rapidly after cryptorchidism, and was related inversely to the PM/ATP ratio, which was also observed in the developing testis. This study demonstrates that MR spectroscopy monitors the cell types present in the rat testis rather than its metabolic status.

Adenosine Triphosphate↗

Global transcriptome analysis of the C57BL/6J mouse testis by SAGE: evidence for nonrandom gene order.

BACKGROUND: We generated the gene expression profile of the total testis from the adult C57BL/6J male mice using serial analysis of gene expression (SAGE). Two high-quality SAGE libraries containing a total of 76 854 tags were constructed. An extensive bioinformatic analysis and comparison of SAGE transcriptomes of the total testis, testicular somatic cells and other mouse tissues was performed and the theory of male-biased gene accumulation on the X chromosome was tested. RESULTS: We sorted out 829 genes predominantly expressed from the germinal part and 944 genes from the somatic part of the testis. The genes preferentially and specifically expressed in total testis and testicular somatic cells were identified by comparing the testis SAGE transcriptomes to the available transcriptomes of seven non-testis tissues. We uncovered chromosomal clusters of adjacent genes with preferential expression in total testis and testicular somatic cells by a genome-wide search and found that the clusters encompassed a significantly higher number of genes than expected by chance. We observed a significant 3.2-fold enrichment of the proportion of X-linked genes specific for testicular somatic cells, while the proportions of X-linked genes specific for total testis and for other tissues were comparable. In contrast to the tissue-specific genes, an under-representation of X-linked genes in the total testis transcriptome but not in the transcriptomes of testicular somatic cells and other tissues was detected. CONCLUSION: Our results provide new evidence in favor of the theory of male-biased genes accumulation on the X chromosome in testicular somatic cells and indicate the opposite action of the meiotic X-inactivation in testicular germ cells.

Animals↗

Presence of immunoreactive corticotropin-releasing factor in rat testis.

We attempted to demonstrate the presence of immunoreactive (ir) CRF in rat testis by RIA, and by an immunocytochemical technique. The RIA was performed on 27,000 x g supernatants of phosphate buffer extracts of adult rat testes, and revealed a high concentration of irCRF (51 - 74 ng/g testis), with a clear parallelism to the standard curve prepared with synthetic rat CRF. In contrast, the peripheral blood level of irCRF was extremely low (less than 0.05 ng/ml). Immunocytochemical studies of irCRF revealed strong specific staining in the Leydig cells and germ cells in normal adult rat testis. Epididymal spermatozoa also stained positive. Testicular irCRF fluctuated significantly with age. The levels (mean +/- SD) assayed in 10, 20, 60 and 90-day-old rats were 41.6 +/- 4.7, 8.7 +/- 0.2, 55.5 +/- 3.3 and 71.3 +/- 3.4 ng/g testis, respectively (P less than 0.01). The level was drastically reduced after abdominal translocation of a testis (9.6 +/- 1.3 ng/g testis), and after hypophysectomy (16.7 +/- 1.6 ng/g testis) in adult rats. However, neither hemicastration nor unilateral cryptorchidism influenced the irCRF levels in the contralateral testis. These data suggest a local production of irCRF which may play a role in regulation of Leydig cell function and sperm maturation in testis and epididymis.

Animals↗

Apoptosis in testis germ cells: developmental changes in gonadotropin dependence and localization to selective tubule stages.

Recent studies have demonstrated apoptotic DNA fragmentation in the testis of immature rats deprived of gonadotropins. However, the exact cell type undergoing apoptosis during testis development and the age differences of gonadotropin dependence of testis cell apoptosis are unclear. The present study used gel fractionation and in situ methods to quantitate developmental changes of testis cell DNA fragmentation and to localize the specific cell type affected in developing rats with and without treatment with a GnRH antagonist. Apoptotic DNA fragmentation in whole testis was measured in rats between 8-70 days of age. A gradual increase (1.8- to 2.0-fold) in testis apoptotic DNA fragmentation was seen in rats between 16-28 days of age, compared with 8-day-old animals, followed by a decrease in adult animals. To study gonadotropin dependence of testicular apoptosis, serum FSH and, to a lesser extent, LH were suppressed by treatment with a long-acting GnRH antagonist (azaline-B, 250 micrograms/kg body wt, two injections at 2-day intervals). Pretreatment with the GnRH antagonist increased apoptotic DNA fragmentation in rats between 16-32 days of age but not in younger and adult animals demonstrating an age-related change in gonadotropin dependence. To identify the exact testis cell type undergoing apoptosis, in situ analysis of DNA fragmentation was performed. In rats at 16-24 days of age, spermatocytes in selected tubules were found to have increased DNA fragmentation. In contrast, neither Leydig cells nor Sertoli cells were affected. In 32-day-old and adult animals, increased DNA fragmentation was seen in early primary spermatocytes of some tubules. Treatment with GnRH antagonist increased the number of cells with DNA fragmentation as well as percentage of tubules affected. In animals between 16-32 days of age, meiotic spermatocytes were labeled, whereas early spermatids were also labeled in 24- and 32-day-old animals. In adult animals, the level of apoptotic DNA fragmentation was not affected by GnRH antagonist treatment. However, DNA isolated from specific stages of the seminiferous tubules of adult animals showed stage-specific changes of apoptotic DNA fragmentation with 2-fold higher levels found in stages I and XII-XIV compared with stage VIII. In situ analysis of adult testis demonstrated that spermatocytes were the major cell type affected. In conclusion, the present study demonstrated that at least three factors determine the onset of apoptosis of the male germ cells: 1) the developmental stage of the animal; 2) serum levels of gonadotropins, especially FSH; and 3) specific stage of the seminiferous epithelial cycle. The present approach provides the basis for future analysis of the role of gonadotropins and other factors in the regulation of testis cell degeneration in normal and pathological states.

Animals↗

Characterization of unique truncated prolactin receptor transcripts, corresponding to the intracellular domain, in the testis of the sexually mature chicken.

We have examined expression of the chicken PRL receptor (cPRLR) gene in different tissues of the chicken by Northern blot analysis. Most tissues examined (ovary, testis, oviduct, kidney, and fat) possess a prominent full-length (4.6-kb) cPRLR transcript. A larger (11.7-kb) transcript is also detected in ovary, oviduct, testis, and kidney after longer exposure. A unique pattern of cPRLR expression was found in the testis of sexually mature chickens, which have an unusually high abundance of three small transcripts (1.2, 1.7, and 2 kb) in addition to the 4.6-kb transcript found in other tissues. Three domain-specific complementary DNA (cDNA) probes were constructed that correspond to the first and second ligand-binding regions in the extracellular domain and the transmembrane-intracellular domain. With these probes, Northern blot analysis of polyadenylated RNA prepared from the testes of a mature (22-week-old) chicken indicates that the highly abundant (1.2- and 1.7-kb) and less abundant (2.0-kb) cPRLR transcripts in testis hybridize only to the intracellular domain probe. Two types of truncated testis-specific cPRLR transcripts were identified using 5'-RACE (rapid amplification of cDNA ends) analysis of polyadenylated RNA from the testis of a 22-week-old chicken. The predominant truncated cDNA sequence contains the highly conserved box 1 motif [(+)box 1 cDNA] and diverges (at nucleotide 1396) from that of the cPRLR cDNA, just downstream of the transmembrane domain. The other truncated cDNA lacks the box 1 motif [(-)box 1 cDNA], which is replaced by 39 bases that could encode a hydrophobic N-terminus with a putative 5'-untranslated region of 131 bases. Young chickens predominately express the full-length cPRLR messenger RNA (4.6 kb) in the testis. At the onset of sexual maturity, there is a dramatic increase in abundance of the testis-specific (+)box 1 transcript, whereas expression of the full-length cPRLR is depressed. The presence of truncated [(+) or (-)box 1] cPRLR transcripts in the sexually mature chicken testis suggests a complex mechanism of PRL action on gonadal function.

Amino Acid Sequence↗

Action of retinoids on embryonic and early postnatal testis development.

The current study investigates the hypothesis that retinoids have a role in embryonic testis development. The action of retinoids on testis development and the expression of retinoic acid receptors (RAR alpha, RAR beta, RAR gamma) were examined. In embryonic day 13 (E13; plug date = E0) testis organ cultures an RAR-selective agonist and all-trans retinoic acid completely inhibited seminiferous cord formation. In contrast, an RAR alpha-selective antagonist had no effect. RT-PCR demonstrated that RAR alpha messenger RNA (mRNA) was expressed at all developmental time points evaluated, which included embryonic day 14 (E14) through postnatal day 30 (P30). Expression of RAR beta mRNA was present at E15 through P2, whereas RAR gamma mRNA was expressed at E18 through P2. Cellular localization of receptors by immunohistochemistry indicated that RAR alpha was localized to the interstitium at E18 and to the seminiferous cords by P0. RAR beta and RAR gamma were detected in both interstitium and cords at E16 and by E18 were mainly expressed in the cords. At P0 RAR beta and RAR gamma were localized to the germ cell populations. To examine retinoid actions, the growth of P0 testis cultures were investigated. Interestingly, retinol and retinoic acid did not inhibit growth of P0 testis cultures but did inhibit the action of growth stimulators. Retinoic acid inhibited FSH, EGF, and 10% calf serum stimulated growth in P0 testis cultures. The hypothesis tested was that the inhibitory effects of retinoids on P0 testis growth may be mediated through the growth inhibitor, transforming growth factor-beta (TGF beta). The action of retinoids on TGF beta mRNA expression was examined in P0 testis cultures. Retinoic acid stimulated TGFbeta3 mRNA expression within 24 h and increased expression of TGFbeta1 and TGFbeta2 after 72 h. Retinol increased expression of TGFbeta1 and TGFbeta2 but not TGFbeta3 after 72 h of treatment. These observations indicate that retinoic acid can influence seminiferous cord formation and testis growth. The inhibitory actions of retinoids may in part be mediated through increased expression of TGFbeta isoforms.

Animals↗

Pituitary adenylate cyclase-activating polypeptide gene expression regulated by a testis-specific promoter in germ cells during spermatogenesis.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a member of the glucagon-related family of hormones that is widely expressed in various tissues. The PACAP messenger RNA (mRNA) and protein is expressed at high levels in the germ cells of the testis, where it locally activates cAMP-coupled receptors located in the somatic Sertoli cells. The PACAP mRNA expressed specifically in the testis is shorter than the mRNA expressed in hypothalamus and includes 127 nucleotides of novel sequence at the 5'-end, suggesting a different start site of transcription in the testes and the utilization of a tissue-specific promoter. Here we present evidence that a single PACAP gene uses a testis-specific promoter to express a mRNA containing a unique exon located 13.5 kb upstream from the first coding exon. As determined by RT-PCR analysis of testis mRNA, the expression of the first testis-specific exon is relatively specific for the testis, as no PACAP mRNA containing the testis-specific first exon was detected in hypothalamic mRNAs. The promoter for the testis-specific PACAP gene was cloned, and a start site for transcription was mapped by primer extension. The testis-specific promoter sequence directs germ cell-specific expression upon transfection of promoter-transcriptional reporter plasmids to populations of testicular cells in vitro and upon expression of a promoter-reporter transgene in mice. Analyses of PACAP gene expression during the spermatogenic cycle, accomplished by RT-PCR of segments of isolated seminiferous tubules, identified intense expression in the postmeiotic round spermatids during developmental stages I-VIII. These observations establish the existence of a specialized PACAP gene promoter whose activity is highly regulated during the spermatogenic cycle.

Animals↗

Expression of prostaglandin D synthetase during development in the mouse testis.

Prostaglandin D synthetase is expressed relatively highly in the testis and reproductive tract of a number of species, including the mouse. In adult mouse testis, expression is confined largely to the Leydig cells and in this study changes in the expression and localization of prostaglandin D synthetase mRNA during testis development were examined. Initial studies using RT-PCR and isolated testicular compartments indicated that prostaglandin D synthetase expression in the neonatal testis was predominantly within the seminiferous tubules. In situ hybridization studies confirmed that prostaglandin D synthetase mRNA appears to be expressed only in the tubules of neonatal mouse testes and only in the interstitial tissue of the adult testis. TaqMan real-time PCR was used to quantify prostaglandin D synthetase mRNA content during development using an exogenous mRNA as a control standard. Expression per testis decreased after birth to < 10% at day 15 before recovering again by days 25-30. After day 30, expression per testis increased 40-fold during final development to adulthood. Studies using RT-PCR showed that early expression before day 15 was restricted to the tubular compartment, whereas the subsequent increase in expression after day 30 was restricted to the interstitial compartment. Database analysis showed that the 3' end of the prostaglandin D synthetase transcript was subject to alternate splicing. Both splice isoforms were shown by RT-PCR to be present throughout development and without a major change in expression pattern. These results indicate that expression of prostaglandin D synthetase mRNA shifts during development from the tubular compartment of the fetal or neonatal testis to the developing adult Leydig cells, with expression in the Leydig cells increasing markedly after puberty. These changes are similar to those observed for 17beta-hydroxysteroid dehydrogenase type III and may indicate that this developmental process is not uncommon in the testis.

Alternative Splicing↗

Comparative study on glutathione transferases of rat brain and testis under the stress of phenobarbitol and beta-methylcholanthrene.

A comparative study was made on the tissue specific expression of glutathione transferases (GST) in brain and testis after exposure of rat to phenobarbitol (PB) and b-methylcholanthrene (MC). Glutathione transferases, a family of multifunctional proteins are involved in intracellular transport processes and in detoxication of electrophilic xenobiotics by catalyzing reactions such as conjugation, isomerization, reduction and thiolysis. On purification, the yield of GST proteins by affinity chromatography was 39% in testis and 32% in brain. The affinity purified testis GSTs were resolved by chromatofocusing into six anionic and four cationic isozymes, and in brain glutathione transferases were resolved into four anionic and three cationic isozymes, suggesting the presence of multiple isozymes with Yc, Yb, Ybeta and Ydelta in both of them. In testis and brain, these isozymes at identical pI values showed variable functions with a battery of substrates and the cationic isozymes of brain and testis showed identical properties in CHP (cumene hydroperoxide) at pH values of above 7.0. Substrate specificity studies and immunoblot analysis of testis and brain proteins revealed that they play a predominant role in the detoxication of phenobarbitol or beta-methylcholanthrene. Expression of the isozymes in testis and brain on exposure to PB and MC indicated elevated subunit variation. In both testis and brain, Ydelta of pi class was expressed on PB treatment and Yc of alpha class and Ybeta of mu class was expressed in MC treated testis and only Yc was predominantly expressed in MC treated brain. Thus these subunits expression is considered as markers for carcinogenesis and specific to chemical toxicity under phenobarbitol and beta-methylcholanthrene stress.

Animals↗

Society of Uroradiology Award paper. The pars infravaginalis gubernaculi: importance in the identification of the undescended testis.

The gubernaculum, a cordlike structure that extends from the testis to the scrotum and guides the testis in its descent, has a bulbous termination, the pars infravaginalis gubernaculi. We reviewed seven surgically proved cases in which the pars infravaginalis gubernaculi was mistaken for an undescended testis on imaging studies. The studies included sonography (five cases), CT (two cases), and MR imaging (two cases). Identification of the mediastinum testis on sonograms or MR images confirms that a structure is testis and not the pars infravaginalis gubernaculi. In two of these patients, testicular venography was used to further identify the undescended testis. In these two patients, the pampiniform plexus, which is used to identify the presence and position of testis, was located adjacent to the pars infravaginalis gubernaculi. Our experience indicates that the pars infravaginalis gubernaculi can be similar in appearance to the undescended testis on any imaging study. The finding of an apparent cord leading to a testislike structure on caudad sectional imaging does not obviate searching for the testis as far as the renal hila.

Adolescent↗

Effects of unilateral castration and unilateral cryptorchidism of the Holstein bull on plasma gonadotropins, testosterone and testis anatomy.

The effects of unilateral castration (UC) and induced unilateral cryptorchidism (CR) on plasma hormones and testis anatomy were studied in 36 Holstein bulls altered at either 3, 6 or 9 mo of age (n = 12). Plasma hormone concentrations were determined in six samples collected at hourly intervals on d 0, 1, 3, 7, 14 and 30, and then at monthly intervals through 6 mo after gonadal manipulation. Although plasma testosterone (T) showed a transient decrease (P less than .05) immediately after treatment, mean plasma concentrations of luteinizing hormone (LH) and T were unaffected by UC or CR over the 6-mo period (P greater than .05). Both hormones increased (P less than .05) in concentration with advancing age. Plasma follicle stimulating hormone (FSH) concentration was greater (P less than .05) in UC than in intact (IN) bulls overall, while FSH in CR bulls did not differ (P greater than .05) from either group. At slaughter, 11 mo after gonadal alteration, mean testis weight, ratio of testis weight to body weight and mean testis sperm cell numbers were increased (P less than .05) in UC bulls compared with mean testis values in intact (IN) bulls. Unilateral castration increased (P less than .05) seminiferous tubuler diameter and seminiferous epithelial cell height from basement membrane to the border of the lumen, but did not alter the ratio of tubuler to interstitial space within the testis. Seminiferous tubuler diameter and epithelial cell height were increased (P less than .05) in CR compared with IN bulls. Unilateral gonadal alteration at 3 mo of age caused a greater (P less than .05) hypertrophy of the scrotal testis in both UC and CR bulls than alteration at 6 or 9 mo of age. Results indicate that unilateral gonadal disruption is followed by rapid compensation in testis T production, little change in systemic LH and a rapid increase in secretion of FSH in the bull within those ages investigated. Further, UC elicits a greater compensatory hypertrophy than CR and the pituitary-testis endocrine axis is more responsive to alteration at 3 mo than at 6 or 9 mo of age in the bull.

Animals↗