Arginase in bull testis

Bull testis arginase was about 1,000 times purified. The molecular weight was estimated to 120,000 (+/- 250). The enzyme was an anionic protein. Km was determined to be 3.4 mM. According to electrophoretic mobility and affinity to DEAE-cellulose as well as to the immunological properties, the arginase isolated from bull testis may be identified as arginase A1.     

Maldescendus testis

Maldescendus testis is a common congenital abnormality occurring in 2-5% of full-term boys at birth in the Western countries. By 3 months of age, the incidence rate spontaneously reduces to 1-2% in this group. The etiology of the disorder is not known, but normal hypothalamo-pituitary-gonadal axis is usually a prerequisite for normal descent of the testes. Abnormal sexual differentiation is associated with maldescent. However, the majority of boys with maldescended testes show no endocrine abnormalities after birth. Several defects in developmental genes, such as homeobox genes and Insl3, have been described to cause cryptorchidism in mice, and disturbances in the regulation of these genes or their mutations may explain etiology of a large part of human testicular maldescent in the future. Increased degeneration of germ cells can be observed in undescended testes after the first year, and therefore early treatment is recommended. Surgical treatment is the most effective and reliable method to bring testes into the scrotum, but hormone treatment with either hCG or GnRH analogues can be considered, particularly in cases where testes can be palpated in high scrotal position. The efficacy of hormone treatment is less than 20% and depends on the initial location of the testis. Nonpalpable testes rarely descend with hormone treatment. Both surgery and hormone treatment can have untoward effects. Treatment with hCG has been associated with an inflammation-like reaction in the testes and an increased rate of apoptosis of germ cells leading to a reduced adult size of the testes. Vascular complications can occur during surgery, particularly in staged orchidopexies. Men with a history of undescended testis have an increased risk of testicular cancer. Impaired fertility is another long-term risk associated to maldescended testes. Fertility potential may be improved by early treatment. Although our knowledge on cryptorchidism has increased considerably during the last decades, many questions remain to be answered: Is the incidence rate increasing? What is causing maldescent? Do hormones have any role in the treatment?     

Accuracy of estimation of testis weight from in situ testis measures in ram lambs

At a mean age of 93 +/- 5 days and a mean weight of 29 +/- 5 kg, 44 crossbred ram lambs were castrated to evaluate the accuracy of the estimation of testis weight from in situ testis measures (scrotal circumference and testis diameter). Means and standard deviations for testis weight (both testes), scrotal circumference and average in situ testis diameter were 134 +/- 57 g, 20.2 +/- 3.1 cm and 3.7 +/- .7 cm, respectively. Testis weight (W) was predicted from scrotal circumference (C) and average in situ diameter (D(o)) as W=.131C(1.90) D(o)(.88) (R(2)=.949). When adjusted to the same scrotal circumference and in situ diameter, testes of 3/4-Finnish Landrace rams were heavier (P<.05) than testes of 7/8-Dorset rams. However, the additional accuracy obtained by using equations specific to each crossbred group was small.     

Androgen receptors in rat testis

Testosterone (T) and 5α-dihydrotestosterone (17β-hydroxy-5α-androstan-3-one; DHT) are bound to specific cytoplasmic receptors (CR) in 105, 000 × g supernatant fractions of seminiferous tubules from hypophysectomized rats following the intravenous injection of [1, 2-³h]testosterone. CR is clearly different from the testicular androgen binding protein (ABP) by electrophoretic mobility, temperature stability and rate of dissociation of steroid-CR complex, but very similar to the cytoplasmic receptors of epididymis and ventral prostate. Under these labeling conditions, the nuclei of seminiferous tubules also contain radioactive T and DHT bound to protein. These androgen-protein complexes, which can be extracted with 0.4 M ? 1 M KC1, have a sedimentation coefficient of 3–4 S. Binding of radioactive T and DHT to both cytoplasmic and nuclear receptors $\text{in vivo}$ is specific for androgen target tissues and abolished by simultaneous injection of unlabeled T, DHT and cyproterone acetate (1, 2-α-methylene-6-chloro-pregn-4, 6-diene-17α-o1–3, 20-diene-17-acetate), but not by cortisol. It is suggested that receptors for testosterone and DHT in the seminiferous tubules are involved in the mediation of the androgenic stimulus to the germ cells.     

Aromatase in the human testis

Low levels of testicular estrogen synthesis have been reported in a number of species, but the cellular localization has not been unequivocally established. To study aromatase in the human testis, we have combined immunocytochemistry with direct measurement of enzyme activity in the testicular 6μm cryosections. Thus, the functionality of the immunoreaction and its sensitivity can be assessed in quantitative terms. Testes were obtained from immediate autopsy from men aged 18–53 years, from surgery from two patients with prostatic cancer (67 and 74 years) and from two normal children aged 8 months and 3 years at autopsy. Benign testicular sex cord tumors were also examined from two unrelated patients aged 5 and 8 years with gynecomastia and diagnosed with Peutz-Jeghers syndrome. Our results consistently showed low to moderate staining intensity of immunoreactive aromatase in comparison to that of normal human placental cryosections. Immunoreactive aromatase was only present in the interstitial Leydig cells and absent from the Sertoli cells of all normal adult testes showing spermatogenesis. Aromatase activity correlated well with the intensity of the immunostain. However, there was no obvious relationship between the level of aromatase activity and increasing age. Generally higher levels were present in testes of young men (18–22 years). No immunostain in any cell type was detected in one 33-year-old patient with testicular cancer. In the testes of the two normal prepubertal boys, no immunostaining was observed. However, intensely stained Sertoli cells as well as high aromatase activity were observed in the testicular tumors of the patients with Peutz-Jeghers syndrome. Our results suggest that Leydig cells are the source of aromatase in normal men but that Sertoli cells may express this enzyme under abnormal conditions. The combined methods for measuring enzyme activity and immunoreactive aromatase are suitable for application to tissues expressing low levels of aromatase.     

Androgen receptors in rat testis

Testosterone (T) and 5α-dihydrotestosterone (17β-hydroxy-5α-androstan-3-one; DHT) are bound to specific cytoplasmic receptors (CR) in 105,000 × g supernatant fractions of seminiferous tubules from hypophysectomized rats following the intravenous injection of [1,2-³H]testosterone. CR is clearly different from the testicular androgen binding protein (ABP) by electrophoretic mobility, temperature stability and rate of dissociation of steroid-CR complex, but very similar to the cytoplasmic receptors of epididymis and ventral prostate. Under these labeling conditions, the nuclei of seminiferous tubules also contain radioactive T and DHT bound to protein. These androgen-protein complexes, which can be extracted with 0.4 M — 1 M KC1, have a sedimentation coefficient of 3–4 S. Binding of radioactive T and DHT to both cytoplasmic and nuclear receptors $\text{in vivo}$ is specific for androgen target tissues and abolished by simultaneous injection of unlabeled T, DHT and cyproterone acetate (1,2-α-methylene-6-chloro-pregn-4, 6-diene-17α-ol-3,20-diene-17-acetate), but not by cortisol. It is suggested that receptors for testosterone and DHT in the seminiferous tubules are involved in the mediation of the androgenic stimulus to the germ cells.     

Rabbit testis proacrosin: Immunological similarities between testis,sperm proacrosins and the initially formed testis acrosin

Anti-rabbit proacrosin IgG was prepared from goat serum following immunization with a homogeneous preparation of rabbit testis proacrosin. The “auto-activation” products of purified testis proacrosin were separated into 68,000 and 34,000 molecular weight (mol wt) acrosins by Sephadex G-100 column chromatography. Immunodiffusion analysis of testis and epididymal sperm proacrosins and acrosins on agarose gel against goat anti-rabbit testis proacrosin showed immunological identity between rabbit testis and sperm proacrosins and the initial testis acrosin (mol wt 68,000). However, the 34,000 mol wt form of testis acrosin showed weaker reaction with the antibody and only partial identity with the proacrosin and the 68,000 mol wt form of acrosin. These results suggest that there is no major structural difference between testis and sperm proacrosins and between proacrosin and the 68,000 mol wt acrosin, but such a structual change occurs when the 34,000 mol wt acrosin is formed.     

Prepubertal testis tumors

Because prepubertal testis tumors are rare, their management has historically been based on experience with the more common adult testis tumors. However, recent studies highlighting the natural history of these tumors and their response to therapy have resulted in a modern management algorithm that optimizes testicular preservation and minimizes the morbidity of adjuvant therapies. Many prepubertal testis tumors are benign and can be managed with testis-sparing tumor excision. Localized malignant tumors (yolk sac tumors) may be managed with excision alone. Recurrent tumors and metastatic disease can almost always be treated successfully with platinum-based chemotherapy.     

Serum and testis selenium concentrations and testis morphology in ram lambs of varying ages

Serum and testis selenium (Se) concentrations, body and testes weights, seminiferous tubule height and width measurements and percent of tubules containing luminal spermatozoa were determined in Se-treated (SSe) and control (NSe) crossbred ram lambs at 60, 90, 120, 150 and 180 days of age. With IM injections, SSe lambs received 3 mg of Se as selenite and NSe lambs received 0.9% saline at 30-day intervals throughout the study. For each age group, lambs were weighed, jugular vein blood collected and testes removed at the designated age. Serum and testis tissue samples for each lamb were assayed for Se, and testis tissue was also evaluated for histological parameters. For all parameters, only serum Se concentrations were affected (P<0.0001) by Se treatment; however, all other parameters were affected (P<0.0001) by age. For combined groups, mean testis Se concentration (0.33 ppm), testes weights, seminiferous tubule measurements and percent of tubules (82.2) containing luminal spermatozoa were greatest (P<0.05) at 180 days of age, and mean testis Se concentrations were significantly correlated with these testicular parameters. These data lend support to the hypothesis that the increase in concentration of testicular Se to adult concentrations (>0.3 ppm) around the time of puberty is associated with rapid testicular development and production of spermatozoa.     

Immunolocalization of clusterin in the ram testis, rete testis, and excurrent ducts

Clusterin, a glycoprotein that elicits cell aggregation, has previously been isolated from ram rete testis fluid, and has been partially characterized. In experiments reported, we have used monoclonal antibodies against clusterin in combination with indirect immunofluorescence microscopy to investigate the distribution of clusterin in the adult ram testis, rete testis, and excurrent ducts. Tissue blocks (5 mm3) were fixed in periodate/lysine/paraformaldehyde containing 0.1% glutaraldehyde and, after embedding, 5-microM sections were prepared for immunolocalization. In the testis, 2 basic patterns were observed: 1) strong to moderate staining for clusterin in the adluminal region with little staining in the basal region of the seminiferous epithelium and germinal cells; and 2) moderate staining throughout the seminiferous epithelium between germinal cells. In the rete testis, strong clusterin staining was localized intracellularly in the rete epithelial cells, most often associated with the luminal surface. In the epididymis, intracellular clusterin was localized in some principal cells of the caput epididymidis. The luminal surfaces and spermatozoa within the lumen were strongly positive. In the vas deferens, clusterin staining was associated with the luminal surface only. The presence of clusterin was clearly detected in unwashed isolated epididymal spermatozoa, but not in spermatozoa washed with phosphate-buffered saline containing 0.05% Tween 20.     

Local control mechanisms in the testis

The gonads are unique organs in that they harbor the cells of the germline and consequently provide the local environment necessary for the normal development and differentiation of gametes. Since the local requirements for germ differentiation differ considerably from those of somatic cells the structural and physiological organization of the gonad is complex and compartmentalized. An elaborate network of local paracrine interactions between the somatic and gametogenic elements appears to be essential for normal germ cell development in mammals. This is especially true for the testis where meiosis is continuous throughout adulthood and where the spermatogenic cycle of the seminiferous epithelium is strictly controlled in time and space. The present paper reviews briefly the rapidly expanding field of testicular paracrinology. Special emphasis is given to the role of intra- and intercompartmental paracrine communication in the development of the male gamete.     

16-ene-steroids in the human testis

Incubation of human testicular homogenates with [4-¹⁴C]pregnenolone gave substantial amounts of an unknown metabolite within 1 min, reaching plateau values of 17–23% of total radioactivity added within 5 min. Mass spectrometry of the metabolite showed it to be identical to the boar sex pheromone precursor androsta-5,16-diene-3β-ol (ADL). In cell cultures the major source of ADL and its dehydrogenated metabolite androsta-4,16-diene-3-one (ADN) was the Leydig cell. In rat and monkey testicular homogenates 16-ene-synthetase activity, a prerequisite for the synthesis of ADL and ADN, was completely lacking, limiting the presence of 16-androstenes to boars and men. In contrast to boars, however, in the human testis no 5-reductase activity was found and consequently no 5-reduced-16-androstenes, e.g. androstenol (AL, musk like) and androstenone (AN, urine like), known sex pheromones in pigs. As both sex pheromones have been identified in urine, plasma, sweat and saliva of men and (especially hirsute) women we hypothesize that AL and AN are synthesized from ADL via ADN peripherically in tissues rich in 5-reductase, i.e. skin, axillary sweat glands and probably also the salivary glands. So far, there is some evidence that both sex pheromones may have similar functions in humans as in boars.     

Teratoma generation in the testis capsule

Pluripotent stem cells (PSCs) have the unique characteristic that they can differentiate into cells from all three germ layers. This makes them a potentially valuable tool for the treatment of many different diseases. With the advent of induced pluripotent stem cells (iPSCs) and continuing research with human embryonic stem cells (hESCs) there is a need for assays that can demonstrate that a particular cell line is pluripotent. Germline transmission has been the gold standard for demonstrating the pluripotence of mouse embryonic stem cell (mESC) lines(1,2,3). Using this assay, researchers can show that a mESC line can make all cell types in the embryo including germ cells(4). With the generation of human ESC lines(5,6), the appropriate assay to prove pluripotence of these cells was unclear since human ESCs cannot be tested for germline transmission. As a surrogate, the teratoma assay is currently used to demonstrate the pluripotency of human pluripotent stem cells (hPSCs)(7,8,9). Though this assay has recently come under scrutiny and new technologies are being actively explored, the teratoma assay is the current gold standard(7). In this assay, the cells in question are injected into an immune compromised mouse. If the cells are pluripotent, a teratoma will eventually develop and sections of the tumor will show tissues from all 3 germ layers(10). In the teratoma assay, hPSCs can be injected into different areas of the mouse. The most common injection sites include the testis capsule, the kidney capsule, the liver; or into the leg either subcutaneously or intramuscularly(11). Here we describe a robust protocol for the generation of teratomas from hPSCs using the testis capsule as the site for tumor growth.     

S-100 protein in the testis

Summary S-100, a protein originally believed to be unique to the nervous system, has recently been found in extraneural cell types. We report here on the presence of S-100 in the testis, namely in Leydig cells and in lymphatic endothelial cells, using immunohistochemical and immunochemical methods. We show that the protein in the testis is immunologically identical to brain S-100. The S-100-labelled cells in the testis exhibit morphological similarities with other cell types in different tissues known to contain S-100.