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PubMed1994

Bovine testis and human erythrocytes contain different subtypes of membrane-associated Ins(1,4,5)P3/Ins(1,3,4,5)P4 5-phosphomonoesterases.

1. We have purified membrane-associated Ins(1,4,5)P3/Ins(1,3,4,5)P4 5-phosphatases from bovine testis and human erythrocytes by chromatography on several media, including a novel 2,3-bisphosphoglycerate affinity column. 2. The enzymes have apparent molecular masses of 42 kDa (testis) and 70 kDa (erythrocyte), as determined by SDS/PAGE, and affinities for Ins(1,4,5)P3 of 14 microM and 22 microM respectively. 3. The two enzymes hydrolyse both Ins(1,4,5)P3 and Ins(1,3,4,5)P4 and are therefore type I Ins(1,4,5)P3 5-phosphatases [nomenclature of Hansen, Johanson, Williamson and Williamson (1987) J. Biol. Chem. 262, 17319-17326]. 4. On chromatofocusing, the partially purified testicular enzyme migrates as two peaks of activity, with pI values of about 5.8 and 5.5. The erythrocyte enzyme exhibits only the latter peak. 5. The testis 5-phosphatase is labile at 37 degrees C, but its activity can be maintained in the presence of 50 mM phorbol dibutyrate (PdBu). After PdBu treatment, a third form of the enzyme, with pI about 6.2, appears on chromatofocusing, but without change in its Km or Vmax. 6. Consideration of the properties of these enzymes and of the 5-phosphatases from other tissues suggests that type I Ins(1,4,5)P3 5-phosphatases are of two well-defined subtypes. We propose that these be termed type Ia [typified by the testis enzyme: approximately 40 kDa, higher affinity for Ins(1,4,5)P3] and Type Ib [typified by the erythrocyte enzyme: approximately 70 kDa, lower affinity for Ins(1,4,5)P3].

Animals↗
PubMed1997

Cloning of the monocarboxylate transporter isoform MCT2 from rat testis provides evidence that expression in tissues is species-specific and may involve post-transcriptional regulation.

The cDNA for the monocarboxylate transporter MCT2 from rat testis has been cloned and sequenced. The derived protein sequence shows 82% identity with that from hamster. Rat MCT2 has a relative insertion of five amino acids in the N-terminal sequence preceding the first predicted transmembrane segment. MCT2 appears to be less highly conserved between species than MCT1. Using Northern blotting of RNA from rat and mouse tissues, MCT2 message was demonstrated to be abundant in the testis where a smaller, less abundant MCT2 transcript was also present. Low levels of a slightly different-sized transcript were found in rat and mouse liver, and mouse kidney. In hamster, only one-size transcript was detected at relatively high abundance in all the tissues examined. Antibodies were raised against a peptide derived from the extreme C-terminus of rat MCT2, and Western blotting with these detected MCT2 in membrane fractions prepared from rat testis, liver and brain but not those from heart or skeletal muscle. In hamster, MCT2 was detected in liver, heart and testis but not in brain [Garcia, Brown, Pathak, and Goldstein (1995) J. Biol. Chem. 270, 1843-1849]. For both rat MCT1 and MCT2 there were marked differences between the relative abundance of their respective messages and the amount of protein in membrane fractions from different tissues. This suggests that expression of both of these transporters in different tissues may be species-specific and regulated post-transcriptionally. The different-sized MCT2 transcripts may arise from alternative splicing. Starvation of rats for up to 48 h did not lead to any change in MCT1 or MCT2 expression in the liver, as determined by either Northern or Western blotting.

Amino Acid Sequence↗
PubMed1999

Emerging trends in the management of the impalpable testis.

BACKGROUND: The management of the impalpable undescended testis is controversial. The study examines emerging trends in the management of this problem. METHODS: Two groups of boys were treated consecutively and recorded prospectively from 1974 to 1984 and from 1990 to 1998 inclusive. A consistent policy of using the preperitoneal approach for impalpable testis was adopted during both time intervals but during the second study period examination under anaesthesia and diagnostic laparoscopy were introduced to ascertain testicular presence and location. RESULTS: Some 919 boys were treated for cryptorchidism during the study period. Ninety boys in the first group (23 per cent) underwent preperitoneal explorations for impalpable testes. Anorchia was present in 18 and orchidectomy was performed in two boys. Thirty boys in the later group (5 per cent) were diagnosed as having impalpable testes. Fifteen boys underwent successful preperitoneal orchidopexy, anorchia was present in 11 and four underwent orchidectomy, carried out for high intra-abdominal testes. CONCLUSION: Examination under anaesthesia and subsequent laparoscopic assessment for all impalpable testes has reduced the need for preperitoneal exploration for the impalpable undescended testis. In this large series, division of the testicular vessels in order to secure scrotal placement of the testis was required in one instance only.

Age Distribution↗
PubMed2001

Effects of long-term treatment with the anti-androgen bicalutamide on human testis: an ultrastructural and morphometric study.

AIMS: To assess the effects of more than 4 years' treatment with the anti-androgen bicalutamide on human testis by clinical, ultrastructural and morphometric analysis. METHODS AND RESULTS: Two patients (aged 74 and 69 years) with prostate cancer were treated for more than 4 years with bicalutamide 50 mg daily. Clinical characterization and follow-up included luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone and prostate-specific antigen (PSA) measurements and clinical response of the tumours. Due to progression of the disease, patients underwent surgical orchidectomy as a further androgen withdrawal therapy. Testis biopsies were studied by light and electron microscopy and analysed by morphometry. Control samples were obtained from the normal testis of two patients with testicular cancer who underwent orchidectomy. Clinical follow-up showed a good response in the control of tumour growth and serum PSA decreased to < 4 ng/mL; concentrations of serum LH, FSH and testosterone were within the normal range. Testicular morphology of treated patients was unexpectedly well preserved; the organization of seminiferous tubules was normal with all the germ line elements and mature spermatozoa present. In some areas, a net increase of peritubular connective tissue was evident which may be a consequence of the age of the patients. CONCLUSIONS: Long-term bicalutamide (50 mg) treatment appears to have very little impact on testis ultrastructure and sperm maturation, while it is effective in the control of androgen-dependent prostatic tumours.

Aged↗
PubMed1998

Higher thyroid hormone levels in neonatal life result in reduced testis volume in postpubertal bulls.

Thyroid hormones appear to determine adult testis size in rodents by regulating the period of Sertoli cell proliferation in the neonatal period. In the present study, the correlation between neonatal thyroid hormone levels (T3 and thyroxin, T4) and postpubertal testis size in Simental bulls was examined. T3 and T4 levels were measured in blood plasma from 35 calves immediately after their arrival at the AI centre at age 3-6 months. Testis size (height and width) was measured at 12 months of age in the same live animals. A significant negative correlation (r = -0.55; p < or = 0.001) was found between T4 and calculated testicular volume using either the Pearson correlation test or linear regression analysis, while the levels of T3 and testis volume showed a negative correlation, although this did not reach statistical significance (r = -0.20, p < or = 0.05). The results of this study suggest, therefore, that neonatal thyroid hormone levels might have the same effect on testicular size in cattle as they do in rodents.

Animals↗
PubMed2002

TestisBank: an internet-based gene sequence database of the testis.

The testis is a highly transcriptionally active organ with hundreds of genes expressed during different stages of spermatogenesis. Scientists working on the testis are restricted to using two sources for further information on testicular genes, GenBank and MEDLINE. However, these two databases are not completely linked and give only very little information on the cellular type of expression or whether a gene is cloned in other tissues but is also expressed in the testis. We have generated an organ-specific database named TestisBank, which is capable of retrieving a more complete set of genes expressed in the different cell types of the testis. Furthermore, it extends to the epididymis which plays an important role in germ cell maturation. TestisBank is automatically updated to match the current status of sequence entries in the genome databases and it provides the user with an interface convenient to handle and provides useful links related to male reproduction. The TestisBank is publicly available at http://medweb.uni-muenster.de/TestisBank/. The design of the TestisBank may provide a simple model for the development of databases in which molecular and literature data are merged, thereby allowing detailed insight into the contribution of different cells/genes to the complex process of spermatogenesis.

Databases, Bibliographic↗
PubMed1997

The intermediate filament protein nestin occurs transiently in differentiating testis of rat and mouse.

Nestin is an intermediate filament (IF) protein (IFP) which occurs during early developmental stages and during regenerative processes in muscle and neuronal cells. The spatial and temporal localization of nestin in the developing testis of rat and mouse was studied by immunolabeling light and electron microscopy and by immunoblotting. Nestin localization was related to the localization of the other major IFPs specific for this tissue, i.e. cytokeratins, vimentin and desmin. Laminin immunocytochemistry and conventional microscopy were used to identify tissues and cells. With the incipient differentiation of the gonadal anlage, the reaction for nestin was weak in the gonadal ridge, whereas the cells of the mesonephric mesenchyme showed a prominent reaction for this IFP. The nestin-specific reaction in the epithelial mesonephric duct and tubules was weak and disappeared at an early phase of differentiation. With the development of the testis proper, nestin was transiently found in several cell types. Nestin was found as well as vimentin and cytokeratins in the Sertoli cells. In the interstitial cells nestin was found together with vimentin and desmin IFPs, and was most prominent in the differentiating myoid cells. After birth, nestin gradually disappeared from the testicular cells and in the rat at puberty was found only in the endothelial cells of some blood vessels. The abolished nestin synthesis in the testis was confirmed by immunoblotting. These results suggest that nestin is required transiently during the development of the testis and mesonephros. The temporary presence of nestin, and several other IFPs during these phases, coincides with key phases of urogenital sex differentiation. This may imply that the orchestrated synthesis of the IFPs nestin, cytokeratins, vimentin and desmin is likely to be linked with the genes regulating sex differentiation.

Animals↗
PubMed1998

Differential distribution of type IV collagen chains in the developing rat testis and ovary.

The localization of type IV collagen alpha 1-alpha 5 chains in the differentiating rat testis and ovary was studied by immunocytochemistry. The initial formation of the testis and ovary included the appearance of collagen alpha 1/alpha 2(IV) chains in the gonadal blastemas. Upon further differentiation of the epithelia of the gonads alpha 1/alpha 2(IV) chains became localized in all of the respective basement membranes (BMs). The alpha 3, alpha 4 and alpha 5 chains of type IV collagen were not detectable in the prenatal rat testis and ovary. With the postnatal differentiation of the rat testis the alpha 3-alpha 5(IV) chains gradually appeared, and were localized in BMs of the testicular cords and seminiferous tubules, rete cords, myoid cells, surface epithelium, Leydig cells, and in some blood vessels. In the postnatal rat ovary, the alpha 3(IV) chain appeared in the BMs of small cortical follicles whereas the BMs of secondary and more deeply localized follicles were devoid of this chain. The alpha 1/alpha 2(IV) chains were abundant in the theca. A reaction for alpha 3-alpha 5(IV) chains also appeared in the BM of the ovarian surface epithelium and of some blood vessels after birth. The present results show that the alpha 3-alpha 5(IV) chains are not only less widely distributed than the alpha 1/alpha 2(IV) chains but are also synthesized much later in development. The late appearance of the alpha 3-alpha 5(IV) chains shows that the development of the mature testicular and ovarian BMs is a long process and that the time schedule for the synthesis of these chains is different from that of many other extracellular matrix proteins. A careful analysis of the expression of alpha 3(IV) chain may be useful in the further study of the kinetics and regulation of ovarian follicular growth.

Animals↗
PubMed1999

Sex-specific localization of laminin alpha 5 chain in the differentiating rat testis and ovary.

The localization of laminin (Ln) alpha 5, beta 1 and beta 2 chains in the differentiating rat testis and ovary was studied by immunolabeling light and electron microscopy. The initial formation of the male and female gonadal blastemas included an emergence of Ln alpha 5 and beta 1 chains, but not of Ln beta 2 chain. The sexual differentiation of the embryonic male gonadal cords included rapid sex-specific disappearance of the incipient Ln alpha 5 chain. The rete testis cords, in contrast, remained positive for Ln alpha 5 chain. In the postnatal testis, the Ln alpha 5 chain reappeared in Ln beta 1 chain-positive cord basement membranes, which also became positive for Ln beta 2 chain. The differentiating myoid cells also gradually became positive for both Ln alpha 5 and Ln beta 1 chains. In the ovary Ln alpha 5 chain persisted in BMs of the cords throughout the fetal phase. Small and newly formed follicles in the early postnatal rat ovary were also positive for Ln alpha 5 chain, whereas growing and large follicles were negative. During the early postnatal phase, Ln beta 1-chain positive follicular BMs became also positive for the Ln beta 2 chain. Basement membranes of testicular and ovarian surface epithelia contained the Ln alpha 5 chain throughout the study. The blood vessels of the male and female gonad showed differentiation-dependent variation in their reactivity for the Ln alpha 5 and beta 2 chains. The present results show that the Ln alpha 5 chain is an early molecular marker for sexual differentiation, which therefore may be regulated by the testis-determining factors. The results also show that in the early postnatal rat ovary, the follicular basement membranes are heterogeneous in their Ln content, which may offer a means to distinguish different follicular populations from each other and to identify the different stages of follicular growth.

Animals↗
PubMed2000

Differential localization of immunoreactive alpha- and beta-subunits of S-100 protein in feline testis.

This study investigates the differential localization of the alpha-subunit (S100-alpha) and the beta-subunit (S100-beta) of the S-100 protein in the feline testis, using immunohistochemistry with polyclonal antibodies to bovine S-100 protein (S-100) and monoclonal antibodies to bovine S100-alpha and S100-beta. Appreciable differences were observed in the cellular localization of the immunoreactivity of each subunit. S-100 was observed in the Sertoli cells, the epithelial cells of the transitional segment of the seminiferous tubules, Leydig cells and the peritubular cells of the seminiferous tubules, but was not observed in the epithelial cells of straight tubules and the rete testis or in the endothelial cells of blood and lymph vessels. S100-alpha immunoreactivity was localized in Sertoli cells, peritubular cells and the epithelial cells of the terminal segment of the tubules, whereas S100-beta immunoreactivity was localized in Leydig cells. The differential localization of the alpha- and beta-subunits of the S-100 protein in the feline testis suggests that this protein is multifunctional and be useful as an investigative tool in studying feline testis function.

Animals↗
PubMed2002

Development of the blood-testis barrier in the mouse is delayed by neonatally administered diethylstilbestrol but not by beta-estradiol 3-benzoate.

A group of newborn mice were treated with 1 micro g dose-1 individual-1 of diethylstilbestrol (DES) on alternate days, from days 1 to 11 postnatally. Another group of mice were treated similarly with 125 ng dose-1 individual-1 of beta-estradiol 3-benzoate (E2B). The testes were sequentially examined up to 84 days of age using light and electron microscopy. Spermatogenic cells in the DES-treated mice differentiated normally from birth until 17 days of age, when they differentiated into pachytene spermatocytes and remained at this meiotic prophase for the next 10 days approximately. The cells then began to differentiate further, ultimately forming spermatozoa by 49 days of age. Confocal and electron microscopy showed that the blood-testis barrier did not form until 28 days of age in the DES-treated mice, and a delay in the functional maturation of this structure, as the blood-testis barrier, was confirmed by intercellular tracer experiments. The arrest of spermatogenesis at the meiotic prophase may have been attributable to the DES-induced defective formation of the blood-testis barrier. No delay of the blood-testis barrier formation was detected in the E2B-treated mice. Thus, DES and E2B, both of which are known as potent oestrogenic compounds, had different effects on the Sertoli cells.

Animals↗
PubMed2003

Immunohistochemical distribution of S-100 protein and subunits (S100-alpha and S100-beta) in the swamp-type water buffalo (Bubalus bubalis) testis.

The distribution and localization of S-100 protein (S-100) and its subunits (S100-alpha and S100-beta) in the testis of swamp-type water buffalo were investigated using immunohistochemistry. S-100 was detected in the Sertoli cells in the convoluted seminiferous tubules, modified Sertoli cells lining the terminal segment of the seminiferous tubules and in the intratesticular excurrent ducts (straight tubules and rete testis). S100-beta showed the same distribution and localization with that of S-100. However, the cytoplasmic extension of the Sertoli cells in S100-beta staining showed less staining intensity compared with that of S-100. S100-alpha showed a positive staining only in the modified Sertoli cells of the terminal segment of the seminiferous tubule. Endothelial cells of blood vessels were also positive with the proteins while the Leydig and spermatogenic cells showed a negative reaction. The localization of S-100 in the testis of the water buffalo was in parallel with that of other artiodactyls which supports the hypothesis that this protein is a multifunctional protein. S100-beta in the Sertoli cells suggests that this protein is involved in establishing blood-testis barrier. Its presence in the modified Sertoli cells and in the epithelium of the excurrent ducts suggest secretory and absorptive function, respectively. Meanwhile, S100-alpha, which was detected only in the modified Sertoli cells, is involved in the secretory activity of these cells that are related to exocrine function.

Animals↗
PubMed2003

Postnatal hypothyroidism does not affect prepubertal testis development in boars.

Young boars were treated with propiothiouracil to induce hypothyroidism to examine its effects on postnatal testicular development. Treatments with 0.1% 4-propyl-2-thiouracil (PTU) in drinking water started after weaning, at 3 weeks of age and all boars were severely hypothyroid at 6 weeks of age as determined by measuring T3 and T4 in blood plasma. Boars were castrated at different ages up to 20 weeks and their testes used for histological and immunohistochemical analyses. Although small but significant reduction in testis weight was observed from 8 to 12 weeks of age, this was not accompanied by significant difference in testicular volume. By 20 weeks of age, at the beginning of puberty, the differences in testis weights between control and treated groups of boars disappeared suggesting there is no lasting effect of hypothyroidism on postnatal development of boar testis. Immunohistochemical staining was used to determine the presence of molecular markers in both Sertoli and Leydig cells. Again, there were no differences between testes from control and treated boars in the pattern or intensity of immunostaining using antibodies against 3beta-hydroxysteroid dehydrogenase, antimullerian hormone or proliferating cell nuclear antigen (PCNA). Immunostaining with antibodies against PCNA showed interesting results as it was observed that Sertoli cells still express this marker of proliferating cells at 14 weeks of age, later than previously suggested cessation of Sertoli cell proliferation. This study suggests that hypothyroidism in boars does not have similar effects on postnatal testis development as reported in some other species.

Animals↗
PubMed1989

Steroid sulfatase activity in homogenates, microsomes and purified Leydig cells from adult rat testis.

Steroid sulfatase (STS) activity was studied in the Long-Evans rat testis. The rate of dehydroepiandrosterone sulfate (DHA-S) hydrolysis determined in whole testis homogenates was low compared to that of the corresponding microsomal fractions, which was, in contrast, as high as that expressed in homogenates from purified Leydig cells. Such an increment in STS activity between total homogenates and the corresponding microsomes was not observed for the seminiferous tubules. The STS affinity reported for total testicular microsomes (Km = 3.47 +/- 0.54 microM; mean +/- SEM) was of the same magnitude as that previously reported for Leydig cells, but was about 3 times higher than that measured for whole testis homogenate (Km = 10.11 +/- 0.92 microM). In vivo hCG treatment decreased the STS affinity in total testicular microsomes without affecting this kinetic parameter in whole testis homogenate. These data suggest that the steroid sulfatase expressed in total testicular microsomes (activity and regulation by hCG) could be considered as a good index of Leydig cell STS activity.

Animals↗
PubMedDate not supplied

Effects of the Booroola gene FecBB on somatic and germ cells of the fetal testis.

The present study was conducted to compare in homozygous FecBBFecBB(BB) Booroola and ++ male fetuses, the body and the testicular growths and the tissular and cellular compositions of the testis between 60 and 140 days of gestation. To eliminate differences in growth due to uterine environment, single embryos have been transferred in recipient Mérinos d' Arles ewes. At 60 and 100 days of gestation, the body masses of BB fetuses were significantly lower than those of ++ foetuses (11 and 13%; P = 0.05); but their testis masses of their total contents of somatic (Leydig or Sertoli) cells did not differ significantly whatever the fetal age. At 100 and 140 days of gestation, testis and body masses were significantly correlated without difference between BB and ++ genotypes. In conclusion, the presence or absence of homozygous gene FecBB does not induce significant differences in somatic or germ cell composition of the testis between 60 and 140 days of gestation.

Animals↗
PubMed1984

Depending on the dose 6-OHDA stimulates or inhibits the testis of immature rats.

The effect of the neurotoxic drug, 6-hydroxy-dopamine (6-OHDA) on the testicular growth, the compensatory testicular hypertrophy, the testicular cytology and the serum testosterone level was studied in immature rats. Unilateral injection of 33 micrograms of 6-OHDA beneath the testicular capsule in animals with two testes did not alter the weight of the gonads but promoted the development of both testes. Unilateral injection of 333 micrograms of 6-OHDA resulted in a decreased weight and severe atrophy of the treated gonad, while a significant weight gain of the contralateral testis occurred. In hemicastrated rats 33 micrograms of 6-OHDA into the remaining testis caused an enhanced compensatory testicular hypertrophy. When 333 micrograms of 6-OHDA was administered beneath the capsule of the remaining testis a diminished compensatory testicular hypertrophy occurred but no signs of degeneration could be observed. Data indicate that the testicular adrenergic elements play a role in the testicular growth, in the development of compensatory testicular hypertrophy and in the cytodifferentiation of the immature testis.

Animals↗
PubMed1986

Effect of estradiol benzoate on rat testis and adrenal.

The effects of in vivo administration of estradiol benzoate at a dose of 50 micrograms/rat/day for 15 days on testosterone production of testis and adrenal in vitro were investigated in rats. The data revealed that the androgen synthesis of both the steroidogenic organs were reduced by the treatment. The decrease in androgen production by testis was related to its reduced sperm count and weight. On the contrary, accumulation of cholesterol occurred in treated rat testis indicating inhibition of its steroidogenesis. This data confirm our earlier reports on the direct effect of estrogen on testis.

Adrenal Glands↗
PubMed2001

From brain determination to testis determination: evolution of the mammalian sex-determining gene.

In mammals, sex is determined by an XY male:XX female sex chromosome system in which a male-dominant gene on the Y chromosome (SRY) determines testis formation. Sex chromosomes evolved from an ordinary autosome pair as the Y chromosome was progressively degraded. The Y chromosome has lost nearly all of its 1500 original genes, and those that survived did so because they evolved a critical role in male determination or differentiation. SRY is typical of Y-borne genes. Comparative gene mapping and sequencing shows that SRY arose quite recently as a degraded version of the SOX3 gene on the X chromosome. SOX3 is expressed predominantly in brain, and so is more likely to be a brain-determining than a testis-determining gene. The male-dominant action of SRYmay be an illusion, as its structure suggests that it works by interfering with the action of a related gene, which in turn inhibits testis development. This hypothesis can give a good account of how a brain-determining gene acquired a role in testis determination via differential dosage of SOX3. SRYhas no central role in sex determination and it can be replaced as a trigger and loft, as have many other Y-borne genes in recent evolutionary history. The absence of SRY in two species of the mole vole (Ellobius) suggests that its useful life is already running out.

Animals↗