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PubMed1989

Development of the inhibitory guanine nucleotide-binding regulatory protein in the rat testis.

The functional development of the inhibitory guanine nucleotide-binding regulatory protein (Gi) and anti-diuretic hormone (ADH) activity was investigated in rat testes. Adult (90-day-old), adolescent (40-day-old), prepubertal (23-day-old), and fetal (20.5 days of gestation) testis cells were cultured with 100 ng/ml pertussis toxin for 24 h. The cells were then cultured with human chorionic gonadotropin (hCG), the ADH agonist arginine vasotocin (AVT), or a combination of the two. Testis cells from rats 23, 40, and 90 days of age that were incubated with hCG increased testosterone production when compared with controls. Preincubation of the cells from postnatal rats with pertussis toxin significantly increased hCG-stimulated testosterone secretion when compared to cells preincubated in medium only at all three ages. AVT suppressed hCG-stimulated testosterone secretion, but this suppression was partially reversed in cells from all postnatal ages preincubated with pertussis toxin. Fetal testis cells showed no response to preincubation with pertussis toxin, even when levels were increased to 400 ng/ml or when pertussis toxin treatment was continued throughout the culture period. AVT also had no effect on fetal testis cells. These results indicate that the Gi protein and AVT are not functional in fetal testes but are active from prepubertal stages of development through maturity.

Animals↗
PubMed1989

Quantitative evaluation of the maintenance and development of spermatocytes and round spermatids in cultured tubule fragments from immature rat testis.

Maintenance and development of spermatocytes and round spermatids was studied in an in-vitro incubation system. This system consisted of open tubule fragments from 26-day-old rat testes, obtained after collagenase treatment. The tubule fragments contained Sertoli cells and spermatogenic cells up to and including a small number of early round spermatids. The number of primary spermatocytes and round spermatids in the tubule fragments was estimated using flow-cytometric analysis, immediately after isolation and after 72 h of incubation. In addition, the activity of LDH-C4 in the tubule fragments was measured. After 72 h of incubation, the percentage of spermatocytes was reduced by 70-80%, but the percentage of spermatids was doubled. The total LDH-C4 activity per well was increased 2-3-fold during 72 h of incubation of the fragments. A modest improvement of the culture results was observed when a combination of FSH, insulin, retinol and testosterone was added to the medium. LDH-C4 activity was investigated to see whether it could be used as a quantitative marker of isolated and cultured spermatocytes and spermatids. It was observed that LDH-C4 activity per cell was decreased when spermatocytes and spermatids were isolated and/or incubated at 4 degrees C. However, the cellular enzyme activity returned to control values during subsequent incubation of the cells at 32 degrees C, either in the absence or presence of a protein synthesis inhibitor. Cellular LDH-C4 activity may be influenced not only by temperature, but possibly also by other cell isolation conditions. It is concluded that LDH-C4 activity may not be a reliable quantitative marker for the presence of spermatocytes and spermatids in culture, but should be used in combination with other analytical methods such as DNA estimation and DNA flow cytometry.

Adenosine Triphosphate↗
PubMedDate not supplied

A comparative study of the effects of non-union of testis and epididymis and cryptorchidism on testicular development and spermatogenesis in the rat.

Malunion of testis and epididymis is a congenital anomaly of human undescended testicles which may be of considerable clinical importance. In an experimental model in rat the effects of early disconnection of testis and epididymis (16 days) on testicular development have been testis and puberty. The results are compared with corresponding effects of cryptorchidism and sham-operation. The testes were weighed, and the relative proportions of haploid, diploid and tetraploid cells were quantified by DNA flow cytometry. The histology was evaluated by light and electron microscopy. The non-union operated testes showed a close to normal weight development until about 40 days of age, whereafter a decline occurred. The progression of the maturation division was only slightly reduced at 30 and 37 days of age, but later the spermatogenesis was significantly inhibited. At 58 days of age the maturation division had practically ceased in non-union operated testes. In cryptorchid animals the onset and progression of maturation divisions were almost totally depressed, and only traces of haploid cells were seen in some specimens. The transient increase in tetraploid cells, reflecting primary spermatocytes seen at about 30 days during normal testicular development, was neither depressed in non-union operated nor in cryptorchid animals. This indicates that the major block in spermatogenesis in both situations is at the stage of the primary spermatocytes. At 58 days of age the testicular histology was similar in non-union operated and cryptorchid testicles with many tubular sections showing only Sertoli cells. Scattered and partly degenerated germinal cells were seen in some sections. No histological signs of increased intratubular fluid pressure were detected, suggesting that pressure atrophy is not decisive for the reduced spermatogenesis in non-union operated gonads.

Animals↗
PubMed1986

Morphometric studies of development of the rat testis under physiological conditions and after CdCl2 poisoning.

The morphometrical data of the rat testis were studied between the 1st day and 1.5 year of life under physiological conditions and after intraperitoneal injection of a single dose of cadmium chloride (1.5 mg CdCl2/kg body weight). Under physiological conditions three main period of development occur: between the 1st and 30th day of life, between the 30th and 45th day of life, and after the 45th day of life. The greatest changes after the injection of cadmium were noticed in the period between the 15th and 30th day of life. The periphery of the organ was the first to suffer severe damage.

Animals↗
PubMed1992

Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and adult testis.

BSAP has been identified previously as a transcription factor that is expressed at early, but not late, stages of B-cell differentiation. Biochemical purification and cDNA cloning has now revealed that BSAP belongs to the family of paired domain proteins. BSAP is encoded by the Pax-5 gene and has been highly conserved between human and mouse. An intact paired domain was shown to be both necessary and sufficient for DNA binding of BSAP. Binding studies with several BSAP recognition sequences demonstrated that the sequence specificity of BSAP differs from that of the distantly related paired domain protein Pax-1. During embryogenesis, the BSAP gene is transiently expressed in the mesencephalon and spinal cord with a spatial and temporal expression pattern that is distinct from that of other Pax genes in the developing central nervous system (CNS). Later, the expression of the BSAP gene shifts to the fetal liver where it correlates with the onset of B lymphopoiesis. BSAP expression persists in B lymphocytes and is also seen in the testis of the adult mouse. All of this evidence indicates that the transcription factor BSAP may not only play an important role in B-cell differentiation but also in neural development and spermatogenesis.

Amino Acid Sequence↗
PubMed1987

Effect of embryonic and/or neonatal diethylstilbestrol and allylestrenol treatment on postnatal development of the chick testis.

The synthetic steroid diethylstilbestrol (DES) and the steroid-like allylestrenol (AE) have been used for years in human medicine for the protection of pregnancy. The hazards to the fetus of gestational DES treatment are well established [2, 17, 18]. Knowledge of a similar effect of AE is still fragmentary. Therefore, further studies are required of the after-effects of embryonic and perinatal AE exposure. In our earlier experiments with polypeptide hormones [4, 5, 6] we have observed that perinatal age is a critical period in the maturation of hormone receptors. In this period the presence of hormone induces the development of its specific receptors. The phenomenon is termed hormonal imprinting [4, 5, 6]. During its maturation the receptor is flexible and the presence of non-specific hormones capable of binding to it may alter its normal development Accordingly, even a single hormone injection in the perinatal period may alter the hormone-sensitivity of the target organ.

Allylestrenol↗
PubMed1989

Insulin-like growth factor I in the developing and mature rat testis: immunohistochemical aspects.

The distribution of insulin-like growth factor I (IGF-I; somatomedin C) was mapped in testes of different aged rats by using immunohistochemical techniques. The antiserum used, K 624, has been demonstrated to be specific for human IGF-I, as defined by several criteria. Antibodies to the M1 subunit of ribonucleotide reductase, a key enzyme in DNA synthesis, were used to visualize meiotic and mitotic cells. Cytoplasmic IGF-I-like immunoreactivity as demonstrable during the first two postnatal weeks in spermatogenic cells, in Sertoli cells, and in Leydig cells. The IGF-I-like immunoreactivity decreased in the Sertoli and Leydig cells during the third and fourth postnatal weeks, and in adult rats, only spermatogenic cells showed IGF-I-like immunoreactivity. In mature rat testes, the spermatocytes were strongly immunoreactive. During puberty and adulthood, the spermatogonia expressed subunit M1 ribonucleotide reductase immunoreactivity, whereas no IGF-I-like immunoreactivity could be detected. No extracellular immunoreactivity was observed. We propose that IGF-I and/or IGF-I-like substances, possibly formed by primary spermatocytes, are likely to be involved in differentiation processes, but not in the initiation of cell proliferation in adult testes. The autocrine and/or paracrine action of IGF-I and/or IGF-I-like substances may thus have different action in developing testes than in adult testes. Our results do, however, not allow firm statements about whether IGF-I and related substances exert their actions on Sertoli cells or spermatogenic cells.

Aging↗
PubMed1992

False-negative biopsy for testicular intraepithelial neoplasia.

A routine biopsy of the contralateral testis obtained during orchiectomy for embryonal carcinoma in a 26-year-old patient was negative for testicular intraepithelial neoplasia (TIN; carcinoma in situ of the testis). However, a rebiopsy that was taken because of unexplained elevation of alpha-fetoprotein 15 months later proved to be positive for TIN. Six previously reported cases of false-negative testicular biopsies obtained during a search for TIN are reviewed. In the light of several thousands of biopsies performed world-wide to date, the number of false-negative biopsies is probably very low. Although TIN is obviously not randomly dispersed throughout the testis in all patients, a routine biopsy of the contralateral testicle in patients with testis cancer remains a valuable tool for early detection of bilateral testicular tumors.-cal distribution of TIN in testes removed for this lesion. Their results suggested that after puberty TIN is usually randomly dispersed throughout the testicle. Support for this concept was recently given by Mumperow et al. (1992). These authors examined tumor-bearing testes and they did not find differences in the presence of TIN in biopsies taken from a location close to the tumor and taken from a location distant from the tumor. Thus, one single biopsy is regard to be representative for the entire testis and one biopsy taken after puberty is also assumed to be reliable for predicting whether the testis will ever develop cancer (Berthelsen and Skakkebaek 1981 a). Conversely, if the biopsy is negative for TIN, a future tumor manifestation in the testicle examined is not expected according to this theory (Skakkebaek et al. 1987). Taken together, the concept of TIN would constitute an ideal avenue for the early detection of testis cancer in high-risk populations with the biopsy being a safe means of discriminating between individuals who will or who will not develop testis cancer.

Adult↗
PubMed1978

The development of Schistosoma haematobium in the hamster.

The in vivo development of Schistosoma haematobium in the hamster was studied. Six stages of development were distinguished on the basis of morphological and histochemical criteria. Schistosomula reached the lung (stage 1) on day three post-infection, with maximum concentrations on day nine. Gut formation occurred in the second stage at day 18. In stage 3 'Organogeny' (day 24) males developed one testis and females a narrow uterus. Pairing and the development of males with sperm-containing testes and females with an ovary characterize stage 4 (day 28). In stage 5 (day 53) vitelline follicles begin to develop in the females. The final stage, oviposition, occurred on days 61-63 and was characterized by the appearance of fully developed eggs in the uterus.

Animals↗
PubMed1989

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↗
PubMed1985

Canine cryptorchism and subsequent testicular neoplasia: case-control study with epidemiologic update.

A retrospective study of 2,912 cryptorchid dogs identified 14 breeds with significantly high risk. Among six distinct closely interrelated breed groups (e.g., toy, miniature, and standard poodles), the risk in the smaller breed was always greater than that in the larger relative, suggesting that genetically influenced maldescent could be, in part, related to physical size or the rate of growth of the involved structures. Testicular tumors were diagnosed in 5.7% of the cryptorchid dogs; half had only Sertoli cell tumors, one-third had only seminomas. The relative risk for Sertoli cell tumor or seminoma was not directly related to a familial risk for cryptorchism. Using the health experience of a control population composed of male dogs with anal sac disease (N = 4,184), there is an estimated relative risk of 9.2 in cryptorchid dogs to develop a testis tumor (95% confidence interval, 5.9-14.3) and 4.2 in dogs with inguinal hernia (95% confidence interval, 1.8-9.5). Considering that the anatomical development of the genital tract, testis descent, and tunic relationships in dog are very similar to that in man, and that the associations of cryptorchism and inguinal hernia with testis neoplasms are also similar, the dog should be an excellent model system to further investigate the causes of human cryptorchism.

Animals↗
PubMed1987

Hyperplasia of spermatic cord nerves: a sign of testicular absence.

The comparative histologic study of the spermatic cord in the absence of testis, epididymis-testis separation, and normal development of both testis and epididymis, revealed that there is nerve trunk hyperplasia and hypertrophy in absence of the testis. This finding may greatly aid the diagnosis of testicular absence in the management of impalpable testes.

Cryptorchidism↗
PubMed1988

Autosomal genes involved in mammalian primary sex determination.

Beginning with findings made during the late 1950s and early 1960s, evidence continues to accumulate in support of the hypothesis that the mammalian Y chromosome carries a gene that induces the undifferentiated foetal gonad in XY individuals to develop as a testis. Recently a DNA sequence has been isolated from the human Y chromosome that appears to be the hypothesized Y-linked testis-determining gene, and advances have also been made toward identifying genes that interact with the Y-linked testis-determining (Tdy) gene to initiate testis formation. These loci have been identified in specific stocks of mice carrying the mutant Thp or TOrl allele at the T locus located on chromosome 17, and in crosses involving the transfer of a Y chromosome from two populations of Mus domesticus into the genomes of specific inbred strains of mice. The data in both cases support the hypothesis that there are several loci involved in testis determination and that abnormal interaction of these loci disrupts initiation of testis determination, resulting in development of ovarian tissue in XY individuals.

Animals↗
PubMed1991

[Arteriographic diagnosis of a tumor arising from an undescended testis].

A case with a tumor developed in an undescended testis diagnosed by arteriography is shown. Radiological diagnostic procedures in the diagnosis of cryptorchism are discussed. It is established that selective testicular arteriography is of great value in the preoperative diagnosis of an intra-abdominally located tumor developed in an undescended testis even in the area of modern high resolution non invasive imaging modalities.

Adult↗
PubMed1991

Transgenic mice demonstrate a testis-specific promoter for angiotensin-converting enzyme.

There are two isozymes of angiotensin-converting enzyme (ACE), one produced by somatic tissues and a smaller protein synthesized by developing spermatozoa (testis ACE). To investigate the molecular control of testis ACE, we generated mice transgenic for a construct containing a putative testis-specific ACE promoter linked to the Escherichia coli reporter gene encoding beta-galactosidase. The transgenic mice express beta-galactosidase protein and RNA only within the testis. Histochemical analysis of the transgenic mice shows co-localization of beta-galactosidase protein and endogenous ACE within elongating spermatozoa. These studies demonstrate that transcription of testis ACE is controlled by a strong intragenic testis-specific promoter that is contained within a 698-base pair fragment immediately upstream from the transcription start site of testis ACE. Characterization of the testis ACE promoter may provide insights into the molecular mechanisms controlling cell stage-specific gene expression in the male germ line.

Animals↗
PubMed1989

Quantitation of testicular and somatic cytochromes c in testis and somatic tissues from developing rats.

By combining chromatographic and spectral procedures, simple and quantitative assays for somatic cytochrome c (cyt cs) and testicular cytochrome c (cyt ct) in crude animal tissue extracts were developed. Using this assay procedure, limited developmental studies of cyt ct and cs were performed with tissue extracts of 27-, 58-, and 85-day-old rats. Specific contents of cyt cs in somatic tissues (i.e., micrograms of cyt c/g of tissue) of these three age groups did not show significant variations. However, the amounts of both cyt ct and cs in testis were markedly increased as the rats approached maturity. Increasing cyt ct/cyt cs ratios as the rat developed to maturity suggest that expression of cyt ct is preferentially required for specific function of testis. Application of both molecular biological techniques and this assay (for holo-cyt ct) should be useful to study the overall regulation of the expression of cyt ct in testis.

Aging↗
PubMed1975

Androgen binding protein as a biochemical marker of formation of the blood-testis barrier.

Androgen binding protein (ABP) was measured during postnatal development in normal and irradiated rats to determine whether development of a blood-testis barrier and formation of a continuous lumen from testis to epididymis is correlated with entry of ABP into the caput epididymis. ABP is found in normal testis as early as 14 days postnatally (0.2 pmol/mg), at which time no blood-testis barrier is observed by the peroxidase perfusion technique. Previous findings have shown a close correlation of blood-testis barrier development and lumen formation. Indeed, ABP is not detectable in the epididymis until 18-20 days of age (1.0 pmol/mg) at which time blood-testis barrier formation and lumen development is complete. Whole body irradiation (125 rads) of pregnant rats at 19-20 days of gestation produces male offspring with seminiferous tubules remarkably free of germinal epithelium and containing essentially only Sertoli cells. These Sertoli cell-enriched (SCE) testes produce normal amounts of ABP between 14 and 21 days postnatally. However, between 21 and 30 days of age the specific activity of ABP is significantly higher in the SCE tests (2.8 pmol/mg protein) than in normal testis (0.5 pmol/mg protein). In the SCE testis neither blood-testis barrier development nor lumen formation are complete until 30 days of age, at which time ABP is first detectable in epididymis of the irradiated rat. Thereafter there is a gradual decline of ABP in the SCE testis-and a dramatic increase in the epididymis. NIH-FSH-S-10 (200 mug/rat) injected SC into 14-day-old normal rats stimulated ABP in the testis from control levels of 0.15 pmol/mg to 1.46 pmol/mg within 4 h after injection. However, no ABP was detectable in the epididymis of either the control or the FSH-stimulated rats. These findings suggest that entry of ABP into caput epididymis is an index of blood-testis barrier formation and lumen development.

Androgens↗