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Cadherin-mediated cell-cell adhesion in the testis.

During spermatogenesis, the differentiating germ cells migrate across the seminiferous epithelium while maintaining close contact with the surrounding Sertoli cells via specialized actin-based adherens junctions (ectoplasmic specializations) and intermediate filament-based anchoring junctions (desmosome-like junctions). Although this migration is essential for correct completion of spermatogenesis, the mechanisms that regulate these anchoring junctions are largely unknown, and most of our knowledge of cell-cell adhesion in testis is based on earlier studies in epithelial tissues. In most epithelia, members of the cadherin superfamily play key roles in intercellular adhesion. Cadherins are calcium-dependent cell-cell adhesion molecules mediating numerous homotypic cell-cell interactions. Until recently there has been controversy about the presence and localization of cadherins in the testis, but now there is increasing evidence that various types of cadherins are expressed in this organ, which underscores their importance in testicular functions. Here, we review the expression patterns, regulatory mechanisms and possible roles of the cadherin-mediated cell-cell adhesion in the testis, and particularly during spermatogenesis. Moreover, attention is paid to additional molecular adhesion complexes in the testis, to associated signaling pathways and to cell adhesion-related innovative ways for male contraception.

Adherens Junctions↗

DNA and RNA synthesis by postpubertal undescended testis in vitro.

To study the spermatogenic potential of postpubertal undescended testis, the 3H-thymidine and 14C-uridine incorporations into normally descended and inguinal undescended testes were examined. The results suggest that DNA synthesis by the inguinal undescended testis is remarkably inhibited but, on the contrary, RNA synthesis is not. The cell-proliferative ability of postpubertal undescended testis may be impaired in spite of the retention of cell viability in the testis.

Adult↗

Acute effect of prolactin on ornithine decarboxylase activity in the rat testis.

A study was conducted to evaluate the effect of the acute treatment with prolactin (PRL) on ornithine decarboxylase (ODC) activity in the rat testis. Injection of a single SC dose of ovine PRL to puberal rats resulted in the activation of ODC from whole testis. This effect was maximal at 4 h after injection, and statistically significant at the dose of 500 micrograms. The effect of PRL was confined to the interstitial space; no change was observed in seminiferous tubules. PRL was unable to further increase testicular ODC activity when injected together with a stimulatory dose of human chorionic gonadotropin (hCG). The effect of PRL was mimicked by injection of a single dose of the dopamine antagonist sulpiride, which provoked a ninefold increase in serum PRL levels. In contrast, PRL did not stimulate testicular ODC activity in hypophysectomized rats, either under basal conditions or during treatment with PRL-hCG, indicating the requirement of a functional hypophysis for the expression of PRL action. These results suggest that the stimulation of testicular ODC activity by PRL is a marker of the trophic response of the testis to this hormone, different from the stimulation of steroidogenesis. This activity could be useful for the study of PRL action on the testis as well as of the interaction between PRL and LH at the testicular level.

Animals↗

Symptoms and early management of germinal tumours of the testis.

The symptomatology and delay in diagnosis in a series of 502 germinal tumours of the testis were surveyed. Pain is much more commonly a feature than is usually appreciated and often leads to the condition being treated as inflammatory for much longer than it should be. Most surgical textbooks are not helpful in this regard. Delay in exploring the testis, and, particularly, delay in initiating follow-up therapy after orchidectomy, were associated with a higher stage of disease and poorer results, most clearly seen in nonseminomatous tumours. As the incidence of testicular tumours seems to be rising, a high index of suspicion in any abnormality of the testis in a post-pubertal male is advocated. Exploration through an inguinal incision for any mass or enlargement of the body of the testis not associated with dysuria and pyuria is essential. If a tumour is found, supplementary treatment must be given without delay. Judicious X-ray therapy, surgery and cytotoxic drugs can save most patients. Seminoma, in particular, can be cured, even in the advanced case.

Adult↗

[Management of the impalpable testis; the value of laparoscopy for subsequent treatment].

During a 5-year period between April 1984 and March 1989, we were unable to palpate 62 testes (56 child patients) at the time of diagnosis. An average of 10 months later, 51 testes (48 patients) still remained impalpable preoperatively. Of the two imaging techniques, computed tomography and ultrasonography, the former is rather superior to the latter for verifying the existence of the impalpable testis. However, careful palpation under anaesthesia results in the accurate location of the testis more often than either of those two imaging techniques. On the other hand, as an aid in management of the impalpable testis, laparoscopy offers the most useful information for use in later surgery. This is because we are able to see the spermatic vessel and the vas deferens intraabdominally, in addition to the abdominal testis. Thirty-six patients (39 testes), whose testes remained impalpable even under anaesthesia, underwent laparoscopy. In two cases, we were unable to perform laparoscopy successfully due to failed pneumoperitoneum. However, in all the other cases, the information which could be obtained was fully utilized during subsequent management. Of a total of 37 instances, 8 testes which were abdominal or just canalicular (pendulous) could be recognized. In addition, 4 more were found to have no spermatic vessel in the visual field and a further 4 had a vessel disappearing before reaching the internal ring. In the remaining 21, we were able to detect the spermatic vessel and the vas deferens. At 59 instances of exploratory surgery, a mere 11 testes, among 27 testes, could be fixed to the bottom of the scrotum, resulting in scrotal dimples lasting for several months in 2 cases. Another 14 testes were placed in the upper scrotal region, while 2 were left in the groin region subcutaneously for lack of any alternative site.

Child↗

[Dose outside the treatment field for external irradiation with high-energy X-ray reduction of the dose of the remaining testis in postoperative irradiation of seminoma].

UNLABELLED: This study aimed to decrease the radiation dose to the disease-free testis in postoperative irradiation for seminoma patients. We consider the factors influencing the peripheral dose (PD) of 10MV X-ray radiotherapy to be the distance between the caudal edge of the irradiation field and the measuring point, the size of the therapeutic irradiation field, the thickness of the lead shield laid above and lateral to the disease-free testis, and the thickness of the lateral absorber. MATERIALS AND METHODS: We measured the scattering radiation dose coming from the accelerator head and that due to irradiation volume. We measured these doses using a testicular phantom as the non-diseased testis. RESULTS: Scattering radiation from the accelerator head mainly contributes to PD, whereas the larger the size of the irradiation field the more the scattering radiation from the irradiation volume contributed to PD. PD changed more at the surface of the phantom than at its center. PD at the testicular phantom could be reduced to less than 1% of the therapeutic dose when it was situated more than 5cm distant from the caudal limit of the irradiation field, the lead shield above the testicular phantom was 7.5cm thick, and the lateral lead shield was 2mm thick. CONCLUSIONS: PD is influenced by many factors. It is necessary to clarify the change in PD at the testicular phantom, and it is important to limit the caudal edge of the irradiation field and to lay the lead shield for the attenuation of radiation on the disease-free testis.

Humans↗

The cellular distribution and kinase activity of the Cdk family member Pctaire1 in the adult mouse brain and testis suggest functions in differentiation.

Pctaire1, a member of the family of cyclin-dependent kinases, has been shown to be particularly abundantly expressed in differentiated tissues such as testis and brain. However, very little is known about the cellular and subcellular distribution and function of Pctaire1 protein(s), which is the focus of this study. We show that Pctaire1 encoded two major proteins of M(r) approximately 62,000 and approximately 68,000, found predominantly in testis and brain. Within these two tissues, Pctaire1 was most abundant in the cytoplasm of terminally differentiated cells, notably, the pyramidal neurons in brain and elongated spermatids in testis. Immunoprecipitation experiments further showed that a kinase activity toward myelin basic protein was associated with Pctaire1 in the adult testis and brain and that its activity was potentially regulated through association with regulatory partner(s). These results suggest that Pctaire1 kinase might have an important role in differentiated cells such as postmitotic neurons and spermatogenic cells.

Animals↗

Arterial supply of the human fetal testis during its migration.

PURPOSE: Irrespective of the surgical technique chosen to treat high undescended testis, preservation of an adequate arterial supply for the testis is crucial for successful orchiopexy with maintenance of normal testicular size and texture. To provide an anatomical background for such a procedure, we performed a systematic study on the number and origin of arteries supplying the fetal testis during its migration from the abdomen to the scrotum. MATERIALS AND METHODS: We studied bilaterally 64 testes from 32 fresh human fetuses 13 to 33 weeks after conception. The fetuses were injected through the thoracic aorta with a microvascular silicone rubber red resin to fill in the arterial tree, thereby enabling identification and dissection of all arteries supplying the testes. RESULTS: Of the 64 testes 3 arteries (testicular, deferential and cremasteric) were found in 46 (71.9%), 2 (testicular and deferential) in 15 (23.4%) and 4 in 3 (4.7%). CONCLUSIONS: The fetal testis is always supplied by at least 2 arteries (testicular and deferential arteries) and by 3 or 4 arteries in nearly 80% of the cases.

Arteries↗

The effects of aging on the seminiferous epithelium and the blood-testis barrier of the Brown Norway rat.

Steroidogenesis and spermatogenesis decrease in aging Brown Norway rats. We therefore hypothesized that there must be accompanying morphological changes taking place in the seminiferous tubules of the aging testis. The testes of Brown Norway rats ranging in age from 3 to 24 months were prepared for light and electron microscopy. To assess the integrity of the blood-testis barrier with age, a lanthanum nitrate study was done. The normal seminiferous tubules present in rats at 3 and 12 months of age were largely replaced at 24 months by fully regressed tubules that were virtually devoid of germ cells and contained large intercellular spaces. An electron-microscopic study of these regressed tubules showed a complete loss of cyclical variations of the organelles of the Sertoli cells. The nucleus was more irregularly shaped and was present at various levels in the epithelium. The endoplasmic reticulum was a loose, vesiculated network that was unlike the elaborate, tubular, anastomotic network noted in young animals. The lysosomes were large, oddly-shaped, and contained lipidic inclusions, in contrast to the distinct membrane-bound lysosomes and dense core bodies found in the young animals. Adjacent Sertoli cell processes encompassed large, empty intercellular spaces, possibly occupied previously by germ cells. The typical Sertoli-Sertoli junctions of the blood-testis barrier in the young animal were rarely seen at 24 months and were replaced by focal contact points, usually between three Sertoli cell processes. In the aged animals, lanthanum nitrate permeated the basal and adluminal compartments, extending between Sertoli cell processes and entering the intercellular spaces and lumen. In summary, during aging, there is a breakdown of the blood-testis barrier, and there are striking changes in the appearance of Sertoli cells. These results suggest a possible intrinsic limitation that prevents stem cells from renewing themselves, whether because of a degeneration of immunological origin or because of a lack of Sertoli cell support.

Aging↗

The permeability of the microvasculature of the perfused rat testis to small hydrophilic substances.

The permeability-surface area products (PS) for sodium, Cr-EDTA, and cyanocobalamine (CoB12) have been determined in isolated perfused rat testes, using the single-passage multiple tracer technique, with albumin as the reference tracer. The validity of using albumin was established from its recovery in the perfusate leaving the testis, which was 98.73+/-0.48% of that for Cr-labeled red cells. The PS values obtained for Na, Cr-EDTA, and CoB12 were correlated with perfusate flow, both below and above levels that were equivalent to normal rates of blood flow in the testis (0.3 mL/[g x min]). The values found at the highest flow rates obtained (between 2.7 and 3.5 mL/[g x min]) were 2230+/-240 microL/(g x min) (n = 8) for sodium, 1460+/-140 microL/(g x min) (n = 7) for Cr-EDTA, and 850+/-80 microL/(g x min) (n = 7) for CoB12. These values are similar to those reported at equivalent flow rates for heart muscle and greater than those reported for skeletal muscle, both of which have unfenestrated capillaries similar to testis, but are less than the values for pancreas and salivary gland, which have fenestrated capillaries and are similar to most other endocrine tissues. However, the permeability coefficients for these markers in the testis (calculated using published values for the surface area of the testicular microvasculature) appear to be considerably greater than for any other tissue studied thus far. By extrapolating extraction values, either linearly or logarithmically, to obtain maximal values for PS for Cr-EDTA and CoB12, and comparing the ratio of these PS area values with the ratio of the diffusion coefficients of these molecules, it can be calculated that the equivalent pore radius for the testicular endothelium is between 5 and 6 nm, comparable to those calculated for other nonfenestrated endothelia.

Albumins↗

Expression of betaB(2)-crystallin mRNA and protein in retina, brain, and testis.

PURPOSE: To evaluate the expression of betaB(2)-crystallin mRNA and protein in rat, bovine, and human nonlens and nonocular tissues. METHODS: betaB(2)-crystallin mRNA levels were detected by RT-PCR. betaB(2)-crystallin protein was purified from rat and bovine tissues by FPLC chromatography. FPLC fractions were analyzed by immunoblotting. The identity of betaB(2)-crystallin protein, isolated from the retina, was confirmed by protein microsequencing. RESULTS: betaB(2)-crystallin transcript was detected in rat brain, rat testis, and human retina by RT-PCR. betaB(2)-crystallin transcript was not found in rat lung, heart, ovary, spleen, thymus, kidney, and liver or in human brain and testis. betaB(2)-crystallin protein was partially purified from and its identity confirmed in rat brain, rat testis, and bovine retina. The bovine retinal protein was further confirmed to be authentic betaB(2)-crystallin by protein microsequencing. CONCLUSIONS: These results establish that betaB(2)-crystallin mRNA and protein are expressed in tissues outside of the lens and outside of the eye including retina, brain, and testis. Extralenticular and extraocular expression of betaB(2)-crystallin, coupled with its participation in phosphorylation pathways, suggests that it has nonrefractive functions in these tissues.

Animals↗

Morphology of testis and epididymis in an ethanol-drinking rat strain (UChA and UChB).

The objective of the present study was to analyze the prospective alterations of the testis and epididymis in a defined strain of alcoholic rats in order to contribute to our understanding of the effects of chronic alcoholism on reproduction. The testis and epididymis of the animals were submitted to morphological analysis by macroscopy, light microscopy and electron microscopy and to morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of testis and epididymis weight, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of the testis and epididymis and in the hypothalamus-pituitary axis of the UCh rats.

Alcoholism↗

Studies on relationship between testicular capsule and sperm transport in rat testis.

AIM: In SD rats, histological changes in the testis were observed after bilateral capsulotomy (of the tunica albuginea) in order to investigate the physiological role of the testicular capsule on sperm transport. METHODS: Bilateral longitudinal capsulotomy was devised to disrupt the capsular contractile function. With this technique, only the tunica vaginalis and tunica albuginea were slit open, leaving the tunica vasculosa intact to embrace the underlying testicular parenchyma. After capsulotomy, the structural changes in the seminiferous tubules, the transitional distal seminiferous segment, and the rete testis were observed. RESULTS: In the capsulotomized testis, there was sperm retention at the transitional seminiferous segment and progressive degenerative changes in seminiferous tubules. CONCLUSION: The results clearly indicated that an intact testicular capsule was required for normal sperm transport from the seminiferous tubules into the rete testis. This is the first attempt to study the physiological role of the testicular capsule in intact animals.

Animals↗

The effect of cardiac arrest on the blood-testis barrier to albumin, tumor necrosis factor-alpha, pituitary adenylate cyclase activating polypeptide, sucrose, and verapamil in the mouse.

Impotence commonly occurs after events such as acute myocardial infarction, coronary bypass, head trauma, and cerebral bleeding, including subarachnoid hemorrhage. We hypothesize that the hypoxia accompanying these events could damage the blood-testis barrier (BTB) and so cause testicular dysfunction, a possible cause of impotence. We examined the effect of cardiac arrest in mice on testis weight and various aspects of BTB function. Testis weight was decreased by about 24% 12 hours after cardiac arrest but had recovered fully by day 7. The testis/serum ratio for albumin was increased 12 hours after arrest, showing a disruption in the vascular BTB with recovery by 24 hours. The testis/serum ratio for sucrose was not consistently elevated, showing that the Sertoli cell BTB remained intact. The testis/serum ratio for verapamil was increased on day 3 of cardiac arrest, suggesting impaired function of the BTB's p-glycoprotein efflux transporter. Transporters for pituitary adenylate cyclase activating polypeptide and tumor necrosis factor-alpha were not affected by cardiac arrest. These results show that cardiac arrest affects testis weight and some aspects of BTB function. Such changes might have long-term effects on testicular function.

Albumins↗

Expression of neurotrophin receptors in the developing and adult testis.

Nerve growth factor (NGF) and the other members of the family of neurotrophic factors (the neurotrophin) are essential for neuronal development and differentiation. Neurotrophins interact with two types of cell surface receptors: a low-affinity receptor (p75 NGF-R) and a high-affinity tyrosine kinase receptor belonging to the trk proto-oncogene family, both expressed in the nervous system and in certain non-neuronal tissues. Recently, NGF immunoreactivity and mRNA have been detected in the testis of the adult mouse, rat and human. In the present report we demonstrate the expression of p75 NGF-R during early gonadal development, by mesenchymal cells of the embryonic mouse and rat testis. In the embryonic testis p75 NGF-R-positive cells are spread through the interstitial compartment; during postnatal development they become organized in a cellular layer that surrounds differentiating myoid cells of the seminiferous tubule. Our results also show the expression in the peripuberal and adult mouse and rat testis, of an abundant and shorter transcript of 3.2 kb that cross-hybridizes to the receptor mRNA (3.7 kb). This new mRNA species, which appears at the beginning of spermatogenesis, is expressed by pachytene spermatocytes and round spermatids.

Animals↗

Glial cell line-derived neurotrophic factor (GDNF) and its receptors GFRalpha-1 and GFRalpha-2 in the human testis.

Glial cell line-derived neurotrophic factor (GDNF) and its receptors GFRalpha-1 and GFRalpha-2 were found in the human testis during fetal development (15-34 weeks of gestation) and in adult men (51-86 years of age) by means of RT-PCR, immunohistochemistry and Western blot techniques. Gene expression of GDNF could be established in the human testis and immunoreactivity (IR) for GDNF was detectable in Leydig cells, Sertoli cells, some spermatocytes and round spermatids as well as in smooth muscle cells of the wall of arterioles and small arteries. In the adult human testis, Sertoli and Leydig cells showed GFRalpha-1-IR, whereas GFRalpha-2-IR was located exclusively in Leydig cells. Different to man, in the rat GDNF-IR in Sertoli cells was detectable only until postnatal day10, providing evidence for species related variability in the expression of GDNF. These findings suggest a critical role for GDNF during the differentiation of testicular structures and provide evidence for an additional important function in the adult human and rodent testis.

Aged↗

Vitamin E ameliorates aflatoxin-induced biochemical changes in the testis of mice.

AIM: To assess the effect of aflatoxin on biochemical changes in the testis of mice and the possibility of amelioration by vitamin E treatment. METHODS: Adult male albino mice were orally administered with 25 or 50 microg of aflatoxin/animal/day (750 or 1500 microg/kg body weight) for 45 days. The testis was isolated and processed for biochemical analysis. RESULTS: There was a significant, dose-dependent reduction in DNA, RNA, protein, sialic acid contents and the activities of succinic dehydrogenase, adenosine triphosphatase and alkaline phosphatase in the testis of aflatoxin-treated mice as compared to the vehicle control. However, the acid phosphatase activity was significantly increased in the aflatoxin-treated mice. Vitamin E (2 mg/animal/day) treatment significantly ameliorated the aflatoxin-induced changes, except the acid and alkaline phosphatase activities in the high dose group. CONCLUSION: Vitamin E treatment ameliorates the aflatoxin-induced changes in the testis of mice.

Acid Phosphatase↗

Cloning and characterization of a novel isoform of calpastatin in human adult testis.

AIM: To clone a new gene related to human spermatogenesis. METHODS: cDNA probes of embryo and adult testis were used to hybridize the cDNA microarray of adult testis, and the clones of differential hybridization were sequenced and analyzed. RESULTS: A novel isoform of calpastatin exclusively and highly expressed in human adult testis was found. CONCLUSION: A novel isoform of calpastatin expresses in human testis and it is related to spermatogenesis.

Adult↗