Testicular responsiveness to long-term administration of hCG and hMG in patients with hypogonadotrophic hypogonadism

Steroidogenic responsiveness and amelioration of sperm number and motility following long-term intramuscular hCG and hMG administration were evaluated in 18 males with hypogonadotrophic hypogonadism (HH). The patients consisted of 13 patients with isolated gonadotrophin deficiency (IGD) and 5 patients hypophysectomized at an early or middle pubertal period. Basal serum levels of testosterone and 17 beta-estradiol were within prepubertal range in all patients before the treatment. Serum testosterone levels reached the normal adult male levels within 12-24 months of the treatment in only 2 of 7 younger patients and 1 of 6 older patients with IGD, whereas in all hypophysectomized patients serum levels of both testosterone and 17 beta-estradiol increased to the levels found in normal adult males within 6 months of the treatment. The mean peak levels of serum testosterone and 17 beta-estradiol, respectively, during the treatment were 2.1 +/- 0.8 (SD) ng/ml and 10.8 +/- 4.9 (SD) pg/ml in younger patients with IGD, 1.4 +/- 0.9 ng/ml and 9.7 +/- 5.1 pg/ml in older patients with IGD and 6.0 +/- 1.2 ng/ml and 34.2 +/- 14.8 pg/ml in hypophysectomized patients. Quantitative improvement in both sperm density and sperm motility were found in 4 of 7 younger patients, 1 of 6 older patients with IGD and all hypophysectomized patients, but only 3 of hypophysectomized patients (3 of 18 patients) could become fertile.     

Induction of puberty in a patient with hypogonadotropic hypogonadism: effect of sequentially applied hCG and pulsatile GnRH administration

Pulsatile substitution with GnRH appears to be the therapy of choice in patients with Kallmann's syndrome, a well defined type of hypogonadotropic hypogonadism. We tried to simplify the treatment and to limit the subcutaneous GnRH therapy to the period absolutely necessary to induce spermatogenesis. Therefore we applied in sequence first hCG to stimulate testicular growth and second pulsatile GnRH application to induce spermatogenesis. We herein report that with this mode of therapy testicular growth from infantile to adult size and normal spermatogenesis could be achieved. We conclude that pulsatile GnRH application is a new effective therapy of hypogonadotropic hypogonadism which can be simplified considerably by pretreatment with hCG.     

Testicular responsiveness to hCG of the ovine fetus in the last third of gestation

The in vivo and in vitro testicular responsiveness to hCG of hemicastrated lamb fetuses 95-99, 110-118 and 130-141 days of gestational age was studied. Basal plasma testosterone (T) levels were similar at all ages (less than 0.25 ng/ml), while the mean testicular concentrations of dehydroepiandrosterone sulfate (DHA-S), 17 alpha-hydroxyprogesterone (17-OHP) and T were higher in 95- to 99-day-fold fetuses. Plasma T levels and the concentration of T, DHA-S, 17-OHP, androstenedione (A) and cyclic adenosine 3'5'-monophosphate (cAMP) were increased by hCG in the hemicastrated animal at all ages. cAMP and T production by enriched preparations of dispersed interstitial cells from control testes was increased by hCG in all groups. In fetuses pretreated with hCG in vivo the addition of hCG in vitro failed to modify cAMP and T production. 100 micrograms of LHRH to a 130-day-old fetus increased plasma LH and T levels. From these experiments, it is suggested that the low plasma LH and T levels found throughout the last trimester of fetal life reflect a relative lack of endogenous LHRH synthesis and/or release, rather than reduced testicular steroidogenic capacity.     

Small supernumerary marker chromosome (sSMC) derived from chromosome 22 in an infertile man with hypogonadotropic hypogonadism

We describe the first case of a supernumerary inv dup(22)(q11.1) in an infertile male with hypogonadotropic hypogonadism. This case supports the opinion that supernumerary inv dup(22)(q11.1) could play a role in male infertility. We suggest that the breakpoint in the region 22q11.1 and/or fourfold dosage of centromeric/pericentromeric sequences of the chromosome 22 may be the cause of hypogonadotropic hypogonadism resulting in impaired spermatogenesis and infertility in our patient.     

Testicular changes in adult rat following bilateral partial (caput) epididymectomy

Adult Wistar rats were either partial (caput) and bilaterally epididymectomized or bilaterally efferentectomized, as controls of duct obstruction. The effects on testicular germinal epithelium were studied at 7, 9, 11, 13, 15, 17, 19, 21, 23 and 25 days after surgery. No abnormalities were detected in sham-operated animals. Epididymectomized animals showed different levels of alterations with progressive disruption of the seminiferous epithelium, emergence of multinucleated bodies and some tubules obliterated by degenerated cells and cellular debris. Half way through the experiment there were tubules lacking their epithelia, as well as the Sertoli cells. On the 25th day degeneration was so important that is affected not only the epithelium (missing in almost all tubules) but also the tubular morphology. Eventually efferentectomized animals showed a progressive alteration, but its level was much lower than that observed after partial epididymectomy, indicating a possible specific function of the caput epididymidis in the control of testicular function.     

Parturition in beef cows following administration of porcine relaxin at ten days prepartum

Administration of procine relaxin (pRLX) to heifers 5 d prepartum has been reported to expedite parturition. Thirty-eight mature crossbred beef cows were randomly assigned to one of three treatment groups. Control animals (C; n = 13) received an intramuscular (i.m.) injection of 2 ml corn oil and 2 ml i.m. phosphate-buffered saline (PBS) 24 h later; relaxin treated animals (RLX; n = 13) received 2 ml i.m. corn oil and 1.0 mg i.m. pRLX 24 h later; estradiol-relaxin treated animals (E-RLX; n = 12) received 20 mg i.m. estradiol benzoate (EB) and 1.0 mg i.m. pRLX 24 h later. Treatment with pRLX occurred at 272.6+/-0.14 d of gestation. The pRLX had been purified to homogeneity from porcine ovaries collected during late pregnancy and was determined to have >/=3000 U/mg by the mouse interpubic ligament bioassay. Peripheral blood samples were collected from all cows at 0, 4, 8 and 24 h, respective to corn oil or EB administration, and assayed for plasma estradiol-17beta. At 24 h post administration of EB, plasma estradiol-17beta concentrations were 48.0+/-10.5 pg/ml for C and RLX cows and 178.5+/-14.8 pg/ml for E-RLX cows. There were no treatment effects (P>/=0.10) for elapsed time from treatment to parturition (304.2+/-22.4 h), gestation length (285.2+/-0.9 d), calving difficulty score (1.05+/-0.04), calf vigor score (1.05+/-0.04) or calf birth weight (38.0+/-0.88 kg). Additionally, there were no retained placental membranes in any cows. Administration of pRLX intramuscularly to beef cows at 10 d before expected parturition was not effective in inducing premature parturition. Furthermore, the effectiveness of pRLX in inducing parturition was not enhanced by pretreatment with estradiol benzoate.     

Gene expression changes in GABA(A) receptors and cognition following chronic ketamine administration in mice

Ketamine is a well-known anesthetic agent and a drug of abuse. Despite its widespread use and abuse, little is known about its long-term effects on the central nervous system. The present study was designed to evaluate the effect of long-term (1- and 3-month) ketamine administration on learning and memory and associated gene expression levels in the brain. The Morris water maze was used to assess spatial memory and gene expression changes were assayed using Affymetrix Genechips; a focus on the expression of GABA(A) receptors that mediate a tonic inhibition in the brain, was confirmed by quantitative real-time PCR and western blot. Compared with saline controls, there was a decline in learning and memory performance in the ketamine-treated mice. Genechip results showed that 110 genes were up-regulated and 136 genes were down-regulated. An ontology analysis revealed the most significant effects of ketamine were on GABA(A) receptors. In particular, there was a significant up-regulation of both mRNA and protein levels of the alpha 5 subunit (Gabra5) of the GABA(A) receptors in the prefrontal cortex. In conclusion, chronic exposure to ketamine impairs working memory in mice, which may be explained at least partly by up-regulation of Gabra5 subunits in the prefrontal cortex.     

Testicular endocrine function in hamadryas baboons (Papio hamadryas) during chronic emotional stress

Radioimmunoassay was applied to examine the time course of blood plasma testosterone content in 10 adult male hamadryas baboons under repeated immobilization stress (IS). In the first series of experiments, the monkeys (5) were subjected to daily IS for 2 h during 6 days. In the second series, 5 animals were exposed to 2-hour immobilization 3 days prior to the analogous stressful cycle. In response to intermittent IS the first series males demonstrated profound and long-term suppression of blood testosterone that persisted for 2 days after IS cessation. Preliminary application of a stressful stimulus substantially reduced the degree of blood testosterone suppression. Transitory decrease in blood testosterone was observed 2 hours after the outset of each stressful exposure, followed by complete recovery of testosterone blood level after 6 hours. The differences in the time course of testosterone indicate that the response of the pituitary-gonadal system is dependent on the regimen of stressful stimulus application.     

Testicular secretion of conjugated and unconjugated steroids in normal adults and in patients with varicocele. Baseline levels and time-course response to hCG administration

The effects of a single im injection of 10,000 IU of hCG on testicular steroid secretions were studied in 15 normal men and 54 patients with varicocele. Plasma levels of the following steroids were determined by radioimmunoassay: T, DHT, A, DHA, 5Adiol, E1, E2, 5P, 170H5P, 170HP, E1S, E2S, TE1, TE2, DHAS, 5AdiolS and hCG as well as FSH and LH in baseline samples. In normal men, peripheral venous blood was collected at 0, 1, 2, 4, 6, 8, 24, 36, 48, 72 and 96 h after hCG. Three types of steroid response were observed. The first, biphasic with an early peak within 2 h and a delayed one after 24 h, was shown by T, its unconjugated precursors and DHT. The second with an early rise plateauing for 6 h and followed by a 36 h peak characterized the estrogen pattern. Finally DHAS did not demonstrate a significant increase but 5AdiolS fluctuated at higher levels than the baseline ones. A significant correlation was only observed between hCG maximum value and E2 and TE1 maximum relative responses. In the patient group, peripheral blood was collected at 0, 1, 1.5, 2, 4, 6, 8, 10, 24 and 48 h after hCG. Baseline levels of all steroids were similar in the two groups. After hCG the same delayed pattern was observed as in the controls. The early relative response was low for T while A, DHT, E2 and E1S levels decreased. Conversely 170HP was higher until 24 h. Spermatic vein blood was collected before and at 1.5, 24 and 48 h after hCG. Steroid baseline levels in the varicocele side were similar to the other. After hCG, despite very large variations, general steroid patterns were comparable to those observed in peripheral vein blood. The important fall at 24 h of T/170HP and T/170H5P concomitant with that of T/E2 might be related to an inhibition of C17-20 lyase activity. The testicular secretion of all the measured steroids, including E1S, could be demonstrated except for DHAS. From our findings, a biogenetic defect evidenced by the fast response to hCG administration might exist in varicocele patients. In view of these results, the protocol of the hCG test should be improved using more frequent blood sampling and determining, besides T, its precursors and estrogen conjugates.     

Luteal function in jennies following PGF(2alpha) treatment 3 days after ovulation

Native PGF(2alpha) and its analogs have been used in the horse mare to manipulate ovarian activity, primarily as luteolytic agents to induce estrus. Despite numerous studies on the effects of these luteolysins in the mare, to date only a single investigation has been conducted in the jenny. The aim of this study was to evaluate the response of the corpus luteum (CL) to a single dose of PGF(2alpha) given 3 days (72h) after ovulation and to establish the plasma progesterone (P4) profile from pre-treatment to post-treatment ovulation in the Martina Franca donkey. Twenty-two jennies were ultrasonographically monitored and treated 72h after the detection of ovulation with 0.075 mg i.m. of R-cloprostenol. From the day of ovulation until ovulation post-treatment, blood was collected daily for P4 determination by enhanced luminescence immunoassay. All the jennies except one, exhibited behavioral signs of PGF(2alpha)-induced estrus within 4 days of treatment lasting 5.4+/-1.16 days. Post-treatment ovulation was also hastened, reducing the interovulatory interval (9.6 days). In response to treatment, plasma P4 concentrations fell to estrus levels and then remained constant until the next ovulation in all but the non-responding animal. Our findings indicate that PGF(2alpha) treatment on Day 3 post-ovulation causes the functional regression of the CL in the jenny, reflected both by the rapid induction of estrus and ovulation and by an abrupt drop in circulating P4 concentrations.     

Definitive localization of X-linked Kallman syndrome (hypogonadotropic hypogonadism and anosmia) to Xp22.3: close linkage to the hypervariable repeat sequence CRI-S232.

Kallmann syndrome is a genetically heterogeneous disease characterized by hypogonadotropic hypogonadism and anosmia. Six families in which the disorder followed an X-linked inheritance were investigated by linkage analysis. Diagnostic criteria were uniformly applied and included tests for hypogonadotropic hypogonadism and anosmia. Close linkage was found by using the hypervariable repeated sequence CRI-S232 (DXS278) previously mapped to Xp22.3. At a maximum lod score of 6.5, the recombination fraction was calculated as .03. Of 30 fully informative meioses, one recombination between the disease locus and the loci recognized by probe CRI-S232 was observed. When an independent approach is used, these results confirm the X-linked Kallmann syndrome assignment previously made by deletion mapping, and allow definitive localization of the syndrome assignment previously made by deletion mapping, and allow definitive localization of the syndrome to the Xp22.3 region. This opens the way to carrier detection and to the identification of a gene responsible for this disorder.     

Findings in normal rats following administration of gliadorphin-7 (GD-7)

This paper discusses the effects of gliadorphin-7 (GD-7) infusion in rats and contrasts them with those of beta-casomorphin-7 (betaC-7). Both induce FLI in a dose related fashion. Very strong expression in both geniculate nuclei (GN) and the alveus hippocampus follow GD-7 400 microgram and betaC-7 30 microgram/kg BW. GD-7 affects only these three regions while betaC-7affects 45. FLI is prevented by Naloxone 2mg/kg BW in all regions except the GN where it is diminished 60%. betaC-7 causes bizarre behavior beginning 60s after infusion is started. GD-7 causes no behavioral change. These findings suggest GD-7 gains access to brain cells by diffusion through circumventricular organs while betaC-7 passes the BBB by carrier facilitation.