Hydrohysteroid Dehydrogenases – Biological Role and Clinical Importance – Review

Communications engineering / telecommunication

Age-related changes in the steroid-producing cells of rat testis

PURPOSE: Leydig cells are the main source of testicular hormones that control spermatogenesis, the male reproductive tract, and male secondary sexual characteristics. It is now well established that testosterone levels are reduced with ageing suggesting that there must be deficit in the steroidogenic capacity of ageing Leydig cells. In this respect, the present work aims at describing some structural and functional alterations in rat Leydig cells upon ageing.MATERIAL AND METHODS: Light and electron microscopic observations and immunohistochemistry for 3І-hydroxysteroid dehydrogenase (3І-HSD) were used to identify some structural and functional features of the Leydig cells of ageing rats.RESULTS: The routine histological analysis revealed that during aging (21 and 24 month- old rats) the Leydig cells undergo atrophic changes in size rather than reduction in their number and as consequence the interstitium of ageing testes appeared expanded. The immunoexpression of 3І-HSD as a marker of LC steroidogenic activity decreased after 18 months of age and the immunostaining considerably reduced in 24-month-old rats when compared to that in 3-month-old control ones. Ultrastructural study of Leydig cells in ageing rat testes revealed the presence of Leydig cells with intact morphology as well as Leydig cells with different degree of degeneration. An age-related progressive increase in the number of Leydig cells nuclei that exhibit apoptotic changes like chromatin fragmentation and compaction into dense masses leading to nuclear shrinkage was found out.CONCLUSION: The results obtained suggest that effects of ageing on steroidogenesis are mainly due to structural and functional alterations in Leydig cells resulting in a decreased testosterone production.Scripta Scientifica Medica 2013; 45(3): 32-35

Expression of testicular angiotensin-converting enzyme in adult spontaneously hypertensive rats.

Recent studies demonstrated that one isoform of angiotensin-converting enzyme named testicular or germinal (tACE) is localized in postmeiotic male germ cells and is essential for fertilizing ability of spermatozoa. Hypertension in spontaneously hypertensive rats (SHR) is androgen-dependent and reduction in male gametes is reported in this experimental conditions. Expression of tACE was not studied under conditions of spontaneous hypertension. The aim of this work is to characterize immuno-expression of tACE in the testis of adult (16-week-old) SHR rats in relation to the changes in blood pressure and serum testosterone level. In 82% of adult SHR, the immuno-expression of tACE followed the normal stage-specific pattern. Destructive testicular changes, germ cells depletion have been observed in 18% of 16-week-old SHR and stronger expression of tACE in stages 8-11 compared to controls was detected. As a result stage specificity in SHR was not as evident as in control. No reaction was found in germ cell depleted tubules in which elongated spermatids were absent. Degenerating germ cells exhibited strong immunostaining comparable to that in residual bodies. The blood pressure was significantly higher in SHR and testosterone levels were more than twice but non-significantly elevated. There was no clear correlation between testicular structural changes, blood pressure level values or serum testosterone levels. Expression of tACE in postmeiotic germ cells, specifically altered by SHR, suggested possible involvement of components of renin-angiotensin system in the process of spermiogenesis. Loss of enzyme expression we found in germ cell depleted tubules in SHR is due to absence of corresponding stages of spermatid differentiation. Therefore, tACE can be used as a marker for germ cell depletion due to hypertension and other pathological conditions

Loss and recovery of androgen receptor protein expression in the adult rat testis following androgen withdrawal by ethane dimethanesulfonate.

Androgens are especially important for the maintenance of spermatogenesis in adulthood and the experimental withdrawal of testosterone (T) production by ethane dimenthanesulfonate (EDS) is a valuable tool for studying androgen-dependent events of spermatogenesis. The aim of the present study was to investigate the specific changes in immunoexpression of androgen receptor (AR) in the testis in relation to degeneration and regeneration of Leydig cell (LC) population and seminiferous epithelium. Immunohistochemistry for AR and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) as well as TUNEL assay for apoptosis were performed on testicular sections of control and EDS-treated rats. Serum LH and T levels were measured by RIA. Our results revealed a total loss of AR immunoexpression from the nuclei of Sertoli (SCs), LCs and peritubular cells during the first week after EDS administration and that coincided with severe drop in T levels. Two weeks after EDS administration, the AR expression was recovered in these cells but normal stage-specificity in SCs was replaced by uniform intensity of AR immunostaining at all the stages of the spermatogenic cycle. The stage-specific pattern of androgen expression in SCs with a maximum at stages VII-VIII appeared 5 weeks after treatment. LC immunoreactivity for 3beta-HSD at different time points after EDS administration correlated with values of T concentration. The maximal germ cell apoptosis on day 7 was followed by total loss of elongated spermatids 2 weeks after EDS treatment. Regeneration of seminiferous epithelium 3 weeks after EDS administration and onwards occurred in tandem with the development of new LC population indicated by the appearance of 3beta-HSD-positive cells and gradual increase in T production. The specific changes in AR after EDS including their loss and recovery in Sertoli cells paralleled with degenerative and regenerative events in Leydig and germ cell populations, confirming close functional relationship between Sertoli, Leydig and germ cells

Androgen action via testicular arteriole smooth muscle cells is important for leydig cell function, vasomotion and testicular fluid dynamics

Regulation of blood flow through the testicular microvasculature by vasomotion is thought to be important for normal testis function as it regulates interstitial fluid (IF) dynamics which is an important intra-testicular transport medium. Androgens control vasomotion, but how they exert these effects remains unclear. One possibility is by signalling via androgen receptors (AR) expressed in testicular arteriole smooth muscle cells. To investigate this and determine the overall importance of this mechanism in testis function, we generated a blood vessel smooth muscle cell-specific AR knockout mouse (SMARKO). Gross reproductive development was normal in SMARKO mice but testis weight was reduced in adulthood compared to control littermates; this reduction was not due to any changes in germ cell volume or to deficits in testosterone, LH or FSH concentrations and did not cause infertility. However, seminiferous tubule lumen volume was reduced in adult SMARKO males while interstitial volume was increased, perhaps indicating altered fluid dynamics; this was associated with compensated Leydig cell failure. Vasomotion was impaired in adult SMARKO males, though overall testis blood flow was normal and there was an increase in the overall blood vessel volume per testis in adult SMARKOs. In conclusion, these results indicate that ablating arteriole smooth muscle AR does not grossly alter spermatogenesis or affect male fertility but does subtly impair Leydig cell function and testicular fluid exchange, possibly by locally regulating microvascular blood flow within the testis

The testicular form of angiotensin converting enzyme as a marker for human sperm quality assessment

Introduction: Spermatozoa are rapidly changing cellular structures that are highly dependent on their interaction with the environment. These interactions cause fundamental changes in the spermatozoa’s cells and membrane. All physiological changes that a spermatozoon goes through are required for fertilization. One of the proteins that are essential for the physiological processes in the spermatozoon membrane is the testicular angiotensin-converting enzyme (tACE). In human ejaculated spermatozoa, tACE is found on sperm plasma membrane in the head, neck, and midpiece of the tail having an active role in the capacitation and acrosome reaction. Aim: Immuno-histochemical and fluorescent testing of the testicular isoform of the angiotensin-converting enzyme during spermiogenesis and acrosome membrane of spermatozoa. Materials and methods: Testis biopsies from infertile males have used immunohistochemical testing and fixed spermatozoa for the immunofluorescence assay of tACE. Results: The immunohistochemical test showed tACE expression during spermiogenesis and its participation in the stages of spermatid differentiation in the testis. The immunofluorescent test follows the manifestation of tACE in untreated, capacitated, and acrosome-reacting spermatozoa. In the process of capacitation and acrosome reaction, we found considerable dynamics accompanied by a change in the expression of tACE on the sperm membrane. Conclusions: tACE expression during spermiogenesis and its visualization in the acrosome region confirms the active role of the enzyme in the processes of maturation, capacitation, and acrosome reaction, which determines the enzyme as a reliable marker for the selection of quality spermatozoa in assisted reproduction

Autocrine androgen action is essential for Leydig cell maturation and function, and protects against late-onset Leydig cell apoptosis in both mice and men

Leydig cell number and function decline as men age, and low testosterone is associated with all “Western” cardio-metabolic disorders. However, whether perturbed androgen action within the adult Leydig cell lineage predisposes individuals to this late-onset degeneration remains unknown. To address this, we generated a novel mouse model in which androgen receptor (AR) is ablated from ∼75% of adult Leydig stem cell/cell progenitors, from fetal life onward (Leydig cell AR knockout mice), permitting interrogation of the specific roles of autocrine Leydig cell AR signaling through comparison to adjacent AR-retaining Leydig cells, testes from littermate controls, and to human testes, including from patients with complete androgen insensitivity syndrome (CAIS). This revealed that autocrine AR signaling is dispensable for the attainment of final Leydig cell number but is essential for Leydig cell maturation and regulation of steroidogenic enzymes in adulthood. Furthermore, these studies reveal that autocrine AR signaling in Leydig cells protects against late-onset degeneration of the seminiferous epithelium in mice and inhibits Leydig cell apoptosis in both adult mice and patients with CAIS, possibly via opposing aberrant estrogen signaling. We conclude that autocrine androgen action within Leydig cells is essential for the lifelong support of spermatogenesis and the development and lifelong health of Leydig cells.—O’Hara, L., McInnes, K., Simitsidellis, I., Morgan, S., Atanassova, N., Slowikowska-Hilczer, J., Kula, K., Szarras-Czapnik, M., Milne, L., Mitchell, R. T., Smith, L. B. Autocrine androgen action is essential for Leydig cell maturation and function, and protects against late-onset Leydig cell apoptosis in both mice and men