Role of GERM cell apoptosis and serum inhibin b as prognostic markers of fertility in children with undescended testis

Undescended testis is a common congenital abnormality in children that contributes to impaired fertility in adulthood. This work aims at evaluating germ cell apoptosis as well as serum inhibin B as prognostic markers of fertility potential in cryptorchid children aged less than 2 years. Methods: The study included 32 children with unilateral undescended testis, divided into 3 age groups: group 1 [3<6 months], group 2 [6 - < 12 months] and group 3 [12 - 24 months]. Control group included 22 healthy age-matched boys. Serum inhibin B, testosterone, follicle stimulating hormone [FSH] and luteinizing hormone [LH] were measured in all subjects. Testicular biopsies from cryptorchid boys were examined histologically and by immunohistochemistry for Fas expression. Serum FSH and LH were significantly lower in cryptorchid infants in age group 1 compared to controls [p = 0.03 and 0.04 respectively]. This was accompanied by defective transformation of gonocytes into adult spermatogonia. In age groups 2 and 3, inhibin B was significantly lower in cryptorchid boys than their age-matched controls [p = 0.014 and 0.01 respectively], while the very low serum testosterone was not significantly different between patients and controls. An age-related decrease in inhibin B was paralleled with delayed Sertoli cell development in age groups 2 and 3. Fas expression in germ cells increased with increasing age until the age of 18 months, while in older children, it was markedly reduced. Serum Inhibin B may be used as a marker for the integrity of the seminiferous epithelium in unilaterally cryptorchid boys and may be considered a predictive marker for future spermato gene sis, especially when supplemented with histological data on germ cell development and apoptosis

Maternal Serum and Amniotic Fluid Inhibin A Levels in Women who Subsequently Develop Severe Preeclampsia

The purpose of this study was to evaluate whether maternal serum (MS) and amniotic fluid (AF) inhibin A levels are elevated in patients who subsequently develop severe preecalmpsia, and to investigate the correlation between MS and AF inhibin A levels in the second trimester. The study included 40 patients who subsequently developed severe preecalmpsia and 80 normal pregnant women. Inhibin A levels in MS and AF were measured with enzyme-linked immunosorbent assay (ELISA). The MS and AF inhibin A levels in patients who developed severe preeclampsia were significantly higher than those in the control group (both for p<0.001). There was a positive correlation between MS and AF inhibin A levels in patients who developed severe preeclampsia (r=0.397, p=0.011), but not in the control group (r=0.185, p=0.126). The best cutoff values of MS and AF inhibin A levels for the prediction of severe preeclampsia were 427 pg/mL and 599 pg/mL, respectively; the estimated ORs that were associated with these cut-off values were 9.95 (95% CI 3.8-25.9, p<0.001) and 6.0 (95% CI 2.3-15.8, p<0.001). An elevated level of inhibin A in MS and AF at the time of second trimester amniocentesis may be a risk factor for the subsequent development of severe preeclampsia.

Inhibin binding sites in bovine pituitary membranes

Membranes derived from bovine pituitary glands free of the neural lobe were used to investigate the presence of binding sites for inhibin, a glycoprotein produced by the ovarian granulosa cells capable of selectively suppressing FSH secretion from the pituitary gland. Optimal concentration of membranes (400 micrograms prot) and 125I-bovine inhibin (2 nM) were incubated in a medium containing 50 mM Tris-HCl pH 7.4, 0.01 M MgCl2 and BSA 0.01 percent in a final assay volume of 200 microliters at 37 degrees C for different time intervals. Non-specific binding was estimated using unlabelled inhibin in excess. The time course of specific 125I-bovine inhibin (2 nM) binding to bovine pituitary membranes is slow with 50 percent binding at approximately 20 min of incubation and reaching equilibrium at 90 min of incubation. The kinetic analysis shows an apparent pseudo first order association rate constant (Kob) equivalent to 4 x 10(-2) min-1. Following equilibrium with the tracer, a large excess of unlabelled inhibin (1.27 microM) was able to displace 84 percent of the specific binding within 120 min of incubation and 50 percent of the binding at approximately 40 min. The analysis under displacing conditions showed an apparent dissociation rate constant (K2) equals to 1.5 x 10(-2) min-1 and an apparent association rate constant (K1) equals to 1.3 x 10(9) M min-1. Thus, the estimation of the apparent kinetic equilibrium dissociation constant (Kd = K2/K2) of the binding of inhibin to bovine pituitary membranes was 1.2 nM. These results show for the first time the existence of bovine inhibin specific binding sites in bovine pituitary, and also that such a binding can take place in the absence of either gonadal and/or hypothalamic influences. They also contribute to the better understanding of the role of non-steroidal hormones such as inhibin, in the regulation of gonadotrophin secretion

Differential distribution of inhibin in the anatomical regions of monkey stomach.

Inhibin, a 10.7 kD FSH (follicle-stimulating hormone) suppressing prostatic peptide has been shown to be synthesized and localized in stomach specimen of monkey. In vitro incorporation of labelled amino acid (3H-leucine) into inhibin followed by specific immunoprecipitation by antiserum to inhibin demonstrated an in vitro de novo biosynthesis of inhibin by monkey stomach. Moreover, the synthesis of inhibin was found to be maximum in fundic zone of gastric mucosa compared to cardiac and antral zone. This was supported by immunohistochemical study of three anatomically different regions, especially wherein fundic zone showed intense positive staining for inhibin. Furthermore, the above data was supplemented by quantitative study of tissue inhibin content by RIA which revealed that the fundic zone of gastric mucosa has a much higher concentration of inhibin than cardiac and antral region. The relationship of zonal concentration of inhibin to gastric anatomy appears to be a noteworthy observation and may serve as an useful tool in our understanding of gastric metabolism and activity.

Hormonal modulation of biosynthesis of prostatic specific antigen, prostate specific acid phosphatase and prostatic inhibin peptide.

Hormonal modulation of in vitro biosynthesis of three prostatic secretory proteins, prostate specific acid phosphatase (PSAP), prostate specific antigen (PSA) and prostatic inhibin peptide (PIP) by human benign hyperplasia (BPH) tissue was studied. LH and inhibins caused increase in the synthesis of all three proteins whereas FSH enhanced the synthesis of PIP and PSA only but decreased PSAP synthesis. Prolactin and thyroid releasing hormone decreased synthesis of PIP and PSAP. However, PSA synthesis was enhanced by TRH and was decreased by prolactin. Estradiol caused significant increase in PSA and PSAP but no discernible changes in PIP synthesis were noticed. Testosterone caused an increase in PIP, PSA and PSAP. These data indicate that biosynthesis of PIP, PSA and PSAP by BPH tissue is under multihormonal regulation.

Endometrium--an extragonadal source of inhibin.

Using polyclonal antibodies generated against human seminal plasma inhibin (10.5 KDa), immunocytochemical localization was carried out in paraffin embedded tissue sections of human endometrial biopsies obtained at various phases of the menstrual cycle. A positive reaction which indicated the presence of inhibin was characterized by the presence of golden yellow or brown colour in the cytoplasm of epithelial cells that formed the glands as well as the luminal lining. The stromal cells however, showed negative staining. In early proliferative phase, the endometrial glands exhibited weak positive staining for inhibin which gradually increased and was intense in late follicular and early secretory phases. The intensity of the staining although was not diminished in the glandular epithelium in the mid as well as late secretory phases, the number of cells showing positive staining appeared to be reduced. Incubation of endometrial biopsies in vitro with labelled amino acid and immunoprecipitation of newly synthesized protein with specific antibodies to inhibin indicated that endometrium is capable of de novo synthesizing inhibin. The above results suggest that endometrium is an extra ovarian source of inhibin and the possible role of endometrial peptide in sperm fertilizing capabilities as well as in pre and post implantation events is suggested.