Accessory Reproductive Organs Research Articles

Preclinical evaluation for noninvasive reversal following long-term vas occlusion with styrene maleic anhydride in langur monkeys

A preclinical evaluation for reversal through a noninvasive approach following long-term vas occlusion with styrene maleic anhydride (SMA) has been attempted in langur monkeys at the level of semen parameters, sperm functional tests, semen biochemistry, histology and ultrastructure of reproductive organs, hematology and serum clinical biochemistry including antisperm antibodies (ASA), prostate-specific antigen (PSA) and testosterone. Noninvasive reversal through palpation, percutaneous squeezing and electrical stimulation, forced vibratory movements and suprapubic percussion in the inguinal segments and per-rectal digital massage was attempted in seven langur monkeys after 540 days following vas occlusion. The results revealed instant azoospermia reversal on the same day of reversal with impaired sperm quality, which showed gradual improvement and normospermia with normal motility and viability after 60–90 days of reversal. Sperm functional tests, including ultrastructure of spermatozoa, indicative of sterility in the initial ejaculations, reached normalcy after 90–120 days of reversal. The seminal plasma biochemistry indicative of obstructive azoospermia regained a normal pattern after 90–120 days of reversal. The morphology of testes that showed focal degeneration during 540 days of vas occlusion and that of vasa deferentia that showed exfoliation of epithelial cells resumed to normal morphology comparable with control animals after 150 days of reversal. The morphology of the epididymis, seminal vesicle and prostate did not show appreciable changes following vas occlusion and after noninvasive reversal compared with those of control animals. Hematology, serum clinical chemistry, ASA, PSA and testosterone fluctuated within control limits, indicating safety of the procedure at the level of accessory reproductive organs. The results suggest that noninvasive reversal is feasible even after long-term vas occlusion with SMA and is safe without adverse side effects.

Progesterone receptors: various forms and functions in reproductive tissues

The unequivocal role of progesterone in a variety of events like ovulation, mammary gland development, establishment and maintenance of pregnancy etc are well established. Also the data are accumulating on its role in male reproductive events. In vertebrates and humans, the biological activity of progesterone is mediated by two progesterone receptor proteins PR-A and PR-B, that arise from the same gene and are the members of nuclear receptor superfamily of transcriptional factors. Several studies have demonstrated that the blockage of progesterone receptor using antiprogestins impairs folliculogenesis, ovulation, implantation and pregnancy. Progesterone receptor (PR), have also been detected in human spermatozoa. However, unlike the conventional PR, sperm PR was localized on the membrane and showed distinct characteristics in terms of its size. There are data to demonstrate the inhibition of progesterone driven functions such as hyperactive motility, acrosome reaction on neutralization of sperm membrane PR with specific antibodies against PR. Further significant decrease in the % of PR positive spermatozoa was observed in infertile cases as compared to the fertile men. This indicated that PR can serve as the marker to define the fertilizing potential of the spermatozoa. Recently we have also shown that the PR is expressed in human testis. This reinforced that this PR protein is an inherent testicular protein and not a secretion of accessory reproductive organs. This review compiles the major observations on the forms of the progesterone receptor in various reproductive tissues.

Adverse Effects of Environmental Toxicants, Octylphenol and Bisphenol A, on Male Reproductive Functions in Pubertal Rats

It has been proposed that a global decline in sperm counts, semen quality, and several male reproductive disorders are associated with exposure to environmental chemicals. Thus, the present study examined the effects of two estrogenic chemicals, octylphenol (OP) and bisphenol A (BPA), on epididymal sperm counts and sperm motility, luteinizing hormone (LH)-releasing hormone (LHRH)-stimulated plasma LH and steroid hormones, insulin-like growth factor I (IGF-I), and accessory reproductive organs in pubertal male Wistar rats. Fifty-day-old rats in the OP group (n=11) and BPA group (n=11) received daily sc injections of the respective chemical at a dose of 3 mg/kg bw dissolved in 0.2 mL DMSO. Rats in the control group (DMSO group; n=10) received 0.2 mL DMSO alone. After 2 wk of treatment, a jugular blood sample was taken, and, on the next day, a second blood sample was taken 1 h after an sc injection of LHRH (250 ng). After 5 wk of treatment, rats were deeply anesthetized and heart blood was collected. Epididymal sperm motility and sperm head counts were determined. LHRH increased plasma LH to higher levels in all groups, but the increases were significant (p<0.01) in the BPA and OP groups. However, despite higher LH levels after LHRH injection, the incremental responses of testosterone and pro-gesterone in the OP and BPA groups were small compared to those in the DMSO group, which showed a small LH response. After 5 wk of treatment, plasma testosterone levels were significantly (p<0.01) reduced in the OP and BPA groups and this was accompanied by reduced (p<0.05) epididymal sperm counts. However, the chemical-treated groups had high basal progesterone levels. No significant effects of chemicals on sperm motility parameters were noted. The chemical-induced increases (p<0.05) of the weight of ventral prostate gland were coincided with elevated plasma IGF-I levels in the BPA (p<0.05) and OP (p<0.01) groups. The present results demonstrated that OP and BPA can reduce sperm counts resulting from lowered plasma testosterone in male rats just after puberty. The enlarged ventral prostate gland may possibly be associated with increased plasma IGF-I, raising the possibility of a link between these chemicals and prostate diseases because IGF-I has been implicated in the pathogenesis of human prostate cancers.

EFFECTS OF FLUTAMIDE ON SEX MATURATION AND BEHAVIOR OF OFFSPRING BORN TO FEMALE RATS TREATED DURING LATE PREGNANCY

Flutamide, when administered subcutaneously to female rats at doses of 3, 10, or 30 mg/kg/day during late pregnancy (gestational days 16-21), significantly and dose-dependently decreased anogenital distance (AGD) of the male offspring in each dose group compared to controls. Significant delays in preputial separation were found in males at a dose of 30 mg/kg, but body weight gain was not inhibited. Cryptorchidism and absence of the prostate gland and seminal vesicles were found in males at doses > or = 10 mg/kg, and testicular hypoplasia at a dose of 30 mg/kg. Hypospadias was noted in all dose groups and vaginal pouches at doses of > or =10 mg/kg. The effects on the accessory reproductive organs were severe, although the effects on the testes themselves were mild. However, those effects appeared to become more pronounced with growth, as evaluated on Days 30 and 42 and Weeks 16 to 18. Most of these affected animals displayed cryptorchidism. Male offspring exposed to flutamide in utero showed impairments of sexual behavior as adults in a dose-related manner. Number and frequency of mounts with intromissions was markedly decreased in all treated groups as compared to controls. At 10 mg/kg, no mounting with ejaculation was observed, and at a dose of 30 mg/kg, no mounting with intromission or ejaculation was observed. These changes in sexual behavior were closely associated with abnormalities of the external genitalia. Animals with hypospadias did not display mounts with ejaculation. However, F1 males that copulated at a dose of 3 mg/kg had a normal reproductive function. Histological examination of the reproductive organs revealed degeneration of the seminiferous tubules, hypospermatogenesis, and hypoplasia and inflammation of the seminal vesicles and prostate. Serum levels of FSH, LH, and testosterone in these animals were comparable between control and all dose groups. Therefore, the male reproductive dysfunction seen in the present study could not be attributed to abnormal sex hormone levels during maturation, but to possible demasculinization of the brain and progressively delayed dysmorphology of the male genitalia caused by fetal exposure to flutamide.

Temporal effects of mancozeb on testes, accessory reproductive organs and biochemical constituents in albino mice

Mancozeb, a fungicide of ethylenebisdithiocarbamate group was orally administered at 800 mg/kg body weight to male Swiss albino mice for 5, 10, 20 and 30 days. Daily body weight of the mice were recorded. The mice were sacrificed by cervical dislocation after 24 h of terminal exposure of mancozeb. Testes weight decreased significantly in 20 and 30 days mancozeb treated mice. However, there was a significant decrease in the number of spermatogonia, diameter of spermatocytes and spermatids in 20 days and number of spermatids in 10 days mancozeb treated mice. Histologic studies of the testis of the mice treated with mancozeb for long duration revealed spermatogenesis inhibition reflected by significant decrease in the number of spermatogenic cells and sperms, when compared with that of controls. In the mice treated with mancozeb for 20 and 30 days showed significant decrease in the weight of the prostate gland. However weight of Cowper's glands decreased significantly in 30 days mancozeb treated mice. There was significant decrease in the kidney, spleen and liver weight, where as thyroid weight increased significantly in mice treated with mancozeb for 30 days. However, thymus weight increased significantly only in the mice treated with mancozeb for 10, 20 and 30 days. In mice treated with mancozeb for 20 days caused significant decrease in the level of protein and a significant increase in the level of total lipids in the testis. However, there was significant decrease in the level of glycogen in the kidney. In mice treated with mancozeb for 30 days caused significant decrease in the levels of protein and glycogen and significant increase in the level of total lipids in the testis and liver and a significant decrease in the protein, glycogen and total lipids in the kidney. These observed effects of mancozeb on testis and biochemical constituents may be due to hormonal imbalance in any of the stages in the hypothalamo–hypophysial–testicular axis.

Dicofol formulation induced toxicity on testes and accessory reproductive organs in albino rats.

Dicofol formulation induced toxicity on testes and accessory reproductive organs in albino rats.

Puberty is delayed in male growth hormone receptor gene-disrupted mice.

The role of insulin-like growth factor-I (IGF-I) in the initiation of puberty and testicular function is poorly understood. Growth hormone (GH) receptor (R) gene-disrupted mice or GHR gene "knockouts" (GHR-KO) are GH resistant and IGF-I deficient. To assess whether the age of sexual maturation is affected by the absence of IGF-I, various parameters of sexual development including testicular and accessory reproductive organ weights, balanopreputial separation, germ cell development, and intratesticular testosterone levels were determined in normal and GHR-KO mice between the ages of 25 and 60 days. In addition, at 36 days of age, the testosterone response to luteinizing hormone (LH) treatment was assessed in these mice. The results indicate that the balanopreputial separation was delayed 5 days, and a significant increase in the weights of the seminal vesicles (SV) occurred later in GHR-KO mice than in normal animals (between 30 and 35 days and between 35 and 40 days, respectively). Also, the weights of testes and epididymii were significantly reduced in GHR-KO mice. The intratesticular testosterone levels and the testosterone response to LH treatment were attenuated in GHR gene-disrupted mice. Furthermore, elongated spermatids appeared later in the testes of GHR-KO mice than in the testes of normal mice. These results suggest that the absence of IGF-I secretion delays the normal course of sexual maturation in male GHR-KO mice, indicating that IGF-I plays an important role in the initiation of puberty in male mice.

Assessment of reversible contraceptive efficacy of methanol extract of Mentha arvensis L. leaves in male albino mice

The present study was undertaken to assess the reversible contraceptive efficacy of methanolic extract of Mentha arvensis leaves. Aqueous solution of the extract (10 mg per day per mouse) when administered orally to male mice of proven fertility for 20, 40 and 60 days caused inhibition of fertility while maintaining their normal sexual behaviour. With the increase in treatment duration, there occurred a corresponding decrease in the mean weight of testis and accessory organs of reproduction. Sperm concentration, motility and viability in the cauda epididymis were also decreased. Spermatozoa with coiled tails also appeared in the epididymal smear. However, all the induced effects returned to normalcy within 30 days following withdrawal of 60-day treatment. Oral administration of the extract also did not affect the body weight of the mice and their blood cells count, packed cell volume, haemoglobin and blood/serum biochemistry.

Feasibility and potential gains of enhancing the subacute rat study protocol (OECD test guideline no. 407) by additional parameters selected to determine endocrine modulation. A pre-validation study to determine endocrine-mediated effects of the antiandrogenic drug flutamide.

Groups of five male and five female Wistar rats were treated by gavage with 0, 1, 10, and 100 mg flutamide/kg body weight for at least 28 days to investigate whether proposed enhancements to the current subacute rodent OECD test guideline no. 407 could be included into the testing routine, which of the current and/or additional parameters would detect endocrine-mediated effects of flutamide reliably and sensitively, and to provide information on intra-laboratory variability. Two identical studies were performed concurrently. The enhanced protocol requests the additional determination of the specific hormones triiodothyronine, thyroxine, thyroid stimulating hormone, follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol, prolactin, testosterone, corticosterone; of oestrus cyclicity and necropsy of all females in the dioestrus stage; of the number of homogenization-resistant testicular spermatids and the number, motility, viability, and morphology of cauda epididymal spermatozoa; of additional organ weights (pituitary, ovaries, uterus, thyroid, male accessory reproductive organs); and of the histopathology of additional organs (pituitary, epididymides, coagulation glands, pancreas, vagina). From a technical standpoint, it was possible to conduct a study according to the enhanced protocol, however, with substantial additional effort, an increase in costs by some 67%, and logistic problems. In line with the specific pharmacological effect of flutamide, treatment-related changes were mainly found in male rats, while females were hardly affected by 100 mg/kg. In male rats, 100 mg/kg was the maximal tolerated dose resulting in reduced body weight gain, but no or little other effects on clinical, haematological, clinico-chemical, or behavioral parameters, and 1 mg/kg was the no-observed-adverse-effect level. Antagonism of peripheral androgen receptors by flutamide resulted in decreased relative organ weights of male accessory reproductive organs, changes that were reliably detected in both studies at 100 mg/kg, but only in one of both studies at 10 mg/kg. Corresponding histopathological changes were also detected reliably at 100 mg/kg. Antagonism of central androgen receptors by flutamide increased LH and FSH levels. LH stimulation of testicular Leydig cells in turn increased testosterone and estradiol levels. Again, all these changes were detected reliably at 100 mg/kg, but only in one of both studies at 10 mg/kg. Corresponding histopathological alterations (increase of LH- and FSH-secreting cells, Leydig cell hypertrophy) were detected reliably and sensitively at 10 mg/kg. Studies on liver enzymes performed outside the scope of the enhanced protocol showed that flutamide at 100 mg/kg generally induced hepatic enzyme activities, but decreased the activity of the sex-specific testosterone-dependent liver enzyme CYP2C11 in male rats. The laboratory methods employed yielded reliable results, i.e., 93.6% of the quantitative measurements obtained in both studies were in agreement. Doubling the animal number from five to ten per sex and dose does not increase the sensitivity of detection of endocrine-mediated effects above the level already provided by histopathological examination of groups of five animals. Some of the proposed enhancements evaluated (additional organgravimetry and histopathology) were helpful in detecting the endocrine-mediated effects of flutamide reliably, while others did not contribute towards this aim (spermatology resulted in doubtful effects, female cyclicity was not affected, hormone determinations provided mechanistic information). Ongoing testing according to the revised version of the enhanced OECD test guideline no. 407 protocol and using ten compounds interfering with the endocrine system by different mechanisms will result in the identification of the most appropriate enhancements.

Detection of progesterone receptor transcript in human spermatozoa.

The present study, to our knowledge, is the first to demonstrate presence of progesterone receptor (PR) transcript in human spermatozoa. The study shows the presence of low copy number PR mRNA in mature human spermatozoa. The PR transcript in spermatozoa was detected by reverse transcriptase-polymerase chain reaction using primers specific for the hormone binding domain and the DNA binding domain of the conventional uterine PR. Further, the cDNA sequence of the partial PR transcript from spermatozoa was found to be identical to the region spanning nucleotides 2694 to 3230 of the conventional PR full-length cDNA sequence. This study also indirectly suggests that the PR protein indeed is an intrinsic sperm protein and is not acquired through proteinaceous secretions of accessory reproductive organs.

Requirement ofwingless signaling andengrailed action in the development and differentiation of reproductive system inDrosophila

The segment polarity geneswingless (wg) andengrailed (en) have been shown to play important roles in pattern formation at different stages ofDrosophila development in the thoracic imaginai discs. We have studied the patterns of expression of these genes in genital discs from wild type larvae, pupae and pharate adults and also from hetero-allelic mutant combinations of these genes. Our results suggest that these genes play vital roles in the normal development and differentiation of genital discs and gonads. In the absence of normalwg oren functions, the flies showed a complete lack of internal accessory reproductive organs and specific defects in the external genitalia. In addition, the testes in such males were small, rounded and with an abnormal cellular organization, although the ovaries in females appeared normal. Temperature shift experiments using the conditional mutant allele ofwg, (wgIL-114) indicated a requirement ofwg signaling from second instar onwards for normal development and differentiation of the accessory reproductive organs. Using a heat-shock allele (Hs-wg) we also show that the spatially regulated expression ofwg as a pre-requisite for normal development and differentiation. Based on the expression patterns ofen andhedgehog (hh) we suggest that even in the genital disc development and differentiation the action ofen is mediated throughhh.

Epididymal sperm motion as a parameter of male reproductive toxicity: sperm motion, fertility, and histopathology in ethinylestradiol-treated rats

The present study was designed to characterize the effect of ethinylestradiol (EE) on epididymal sperm motion using a computer-assisted sperm analysis system (CASA), and to elucidate the correlation between sperm motion endpoints and other measures including fertility, histopathologic, and endocrinologic endpoints. EE was orally given to adult male rats at a daily dosage of 10 mg/kg for 3 and 5 d, and at daily dosages of 1 and 10 mg/kg for 1, 2, 3, and 4 weeks. Changes in sperm motion were first detected after one week of treatment. Of nine sperm motion parameters, the percentage of motile sperm, velocity, and amplitude of the lateral head displacement (ALH) were decreased in the 10 mg/kg dosing group. Accompanying the decreases in those parameters, the male fertility indices in the 10 mg/kg dosing group were reduced after one week of treatment, and no males in this group could impregnate intact females after 2 weeks or more of treatment. The number of sperm heads in the cauda epididymis in the 10 mg/kg dosing group was reduced to about one-half that in the control group after one week of treatment, whereas the total number of homogenization-resistant advanced spermatids in the testis was not altered and only a slight change was detected in the number and morphology of germ cells in the testis. These results suggest that reduction in the number of epididymal sperm and in sperm motion are not secondary to testicular alteration. However, after 3 weeks of treatment, the number of sperm heads in the testis was drastically reduced with severe atrophy of the seminiferous tubules both in the 1 and 10 mg/kg dosing groups. The profiling of epididymal luminal fluid proteins indicated that two major bands that migrated with molecular weights of about 22 and 23 kDa were weakened and their density was reduced to approximately 70% of the control after 5-d and one week treatments in the 10 mg/kg dosing group. Circulating testosterone declined drastically after 3 d of treatment and remained at undetectable levels with a concomitant decline of circulating LH and FSH, suggesting that EE inhibits testosterone secretion immediately via a negative feedback system, and there follow changes in the accessory reproductive organs including the epididymis. These results indicate that EE affects epididymal spermatozoa before testicular germ cells via a testosterone deficiency, when it is administered at extremely high dosages. The reduction in the sperm motion manifested as decreases in the percentage of motile sperm, ALH, and velocity, is considered to be responsible for the onset of infertility. Sperm motion analysis could be particularly useful for detecting the toxic effects of chemicals that act through the endocrinologic system on the epididymis.

Serotonin in Golden Hamster Testes: Testicular Levels, Immunolocalization and Role during Sexual Development and Photoperiodic Regression-Recrudescence Transition

Serotonin (5-HT) is found in the gonads and accessory reproductive organs of several species. The golden (Syrian) hamster is a seasonal breeder. Exposure of male adult hamsters to short days for 14 weeks results in a severe gonadal regression, while after a photoinhibition period of 22 weeks a spontaneous testicular recrudescence occurs. The aim of this study was to investigate the presence of 5-HT and its major metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the gonads of golden hamsters, its immunolocation and its physiological role in the testis. The influence of age and photoperiod was also analyzed. Hamsters of 23, 36, 46, 60 and 90 days of age were kept in long photoperiod (LP: 14:10 h light/dark), and adult animals were exposed either to LP or to short photoperiod (SP: 6:18 h light/dark) for 14 and 22 weeks. Testicular parenchyma and capsule levels of 5-HT and 5-HIAA increased significantly at ages of 36 and 60–90 days, but decreased markedly during the exposure of adult hamsters to SP for 14 and 22 weeks. Mast cells were found exclusively in the testicular capsule. The testicular number of mast cells increased concomitantly with age, but decreased in adult hamsters exposed to SP. Mast and Leydig cells presented 5-HT-positive immunoreactivity. During sexual maturation as well as during the transfer of adult hamsters from LP to SP, the 5-HIAA/5-HT ratio showed the highest values in active adult animals, indicating that the increase in testicular 5-HT levels in adulthood is accompanied by an augment in 5-HT turnover. In vitro basal and hCG-stimulated testosterone production was significantly inhibited in presence of physiological concentrations of 5-HT. In conclusion, the present studies demonstrate the existence of 5-HT in mast cells and Leydig cells of hamster testes, as well as describe an inhibitory action of this neurotransmitter on gonadal testosterone production. Furthermore, the age-dependent and photoperiodic-related changes detected in testicular 5-HT levels suggest that this neurotransmitter might act as an important local modulator of the action of gonadotropins on steroidogenesis during sexual development and during the photoperiodic regression-recrudescence transition in the golden hamster.

Seminal vesicles are novel sites of luteinizing hormone/human chorionic gonadotropin-receptor gene expression.

The hypothesis that rat seminal vesicles may contain luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptors was tested by means of a number of different techniques. Northern blotting demonstrated that rat seminal vesicles contained multiple LH/hCG-receptor transcripts. In situ hybridization revealed that these transcripts were present primarily in the principal epithelial cells lining the lumen. Western immunoblotting detected proteins, two of which disappeared (80 and 46 kDa) and another of which decreased (30 kDa) after preabsorption of the receptor antibody with excess receptor peptide. Ligand blotting showed that 125I-hCG could bind only to an 80-kDa protein and that this binding was inhibited by coincubation with excess unlabeled hCG. Immunocytochemistry demonstrated that LH/hCG-receptor protein was present primarily in the principal epithelial cells. In conclusion, our data demonstrate that seminal vesicles contain LH/hCG receptors, thus making this previously unsuspected male accessory reproductive organ a potential target of direct regulation by LH.

A physiologically based approach to the study of bisphenol A and other estrogenic chemicals on the size of reproductive organs, daily sperm production, and behavior.

Two chemicals previously shown to have estrogenic activity, bisphenol A and octylphenol, were examined for their effects on accessory reproductive organs and daily sperm production in male offspring of mice fed these chemicals during pregnancy. These chemicals are used in the manufacture of plastics and other products, and have been detected in food and water consumed by animals and people. From gestation day 11-17 female mice were fed an average concentration (dissolved in oil) of bisphenol A or octylphenol of 2 ng/g body weight (2 ppb) and 20 ng/g (20 ppb). The 2 ppb dose of bisphenol A is lower than the amount reported to be swallowed during the first hour after application of a plastic dental sealant (up to 931 micrograms; 13.3 ppb in a 70 kg adult). We found that the 2 ng/g dose of bisphenol A permanently increased the size of the preputial glands, but reduced the size of the epididymides; these organs develop from different embryonic tissues. At 20 ng/g, bisphenol A significantly decreased efficiency of sperm production (daily sperm production per g testis) by 20% relative to control males. The only significant effect of octylphenol was a reduction in daily sperm production and efficiency of sperm production at the 2 ng/g dose. A new approach to studying physiologically relevant doses of environmental endocrine disruptors is discussed, particularly with regard to the development of the reproductive organs, the brain, and behavior.

Chronic administration of 4-tert-octylphenol to adult male rats causes shrinkage of the testes and male accessory sex organs, disrupts spermatogenesis, and increases the incidence of sperm deformities.

The environmental toxicant 4-tert-octylphenol (OP) has been shown to exert estrogenic effects on mammalian cells in culture. Recent findings from our laboratories demonstrate clearly that OP administration disrupts reproductive hormone secretion in the adult male rat, quite likely as a result of estrogenic action. In the present study, we investigated the impact of these or other OP-induced changes on male reproductive tissues. Adult male rats were injected with OP (20 or 80 mg) or estradiol valerate (EV; 0.8 or 8 microg) s.c. in oil three times a week for either 1 or 2 mo. We found that an 80-mg dosage of OP for 2 mo or an 8-microg dosage of EV for 1 or 2 mo greatly reduced sperm numbers and adversely influenced the sizes, weights, and histological structures of the testes, epididymides, ventral prostate glands, seminal vesicles, and coagulating glands. The 80-mg dosage of OP for 1 mo reduced epididymal tubule size to a lesser extent than after 2 mo of treatment. Otherwise, treatment with 80 mg OP for 1 mo, 20 mg OP for 1 or 2 mo, or 0.8 microg EV for 1 mo had little or no effect on the histology of the tissues we examined. Additional evaluation of sperm morphology revealed marked increases in the proportions of head and tail abnormalities from animals that had received 80 mg of OP or 8 microg of EV for 1 mo and 20 mg of OP for 2 mo. The head abnormalities consisted mainly of pin heads, detached heads, and the absence of hooks, while tail abnormalities included mainly broken, coiled, and bent tails. Our results clearly demonstrate that OP can severely reduce the size and/or function of all of the male gametogenic and accessory reproductive organs studied. Moreover, the similarity of these cell and tissue changes between rats treated with OP and those treated with EV further suggests that OP may exert its action in an estrogenic-like manner.

Immunohistolocalization of carbonic anhydrase isozymes (CA-I, CA-II and CA-III) in bovine male reproductive tracts.

Immunohistochemical localizations of carbonic anhydrase isozymes (CA-I, CA-II and CA-III) in bovine male reproductive tracts were studied. In bulls, no immunoreaction was seen after treatment with antibodies to CA-I, -II and CA-III in the testis. Specific staining for CA-III, however, was evident in peritubular cells in interstitial tissue of the testis, epididymis. CA-II activity could be detected in epithelium of the epididymis, ductus deferentis and ampulla ductus deferentis. Especially, a strong reaction for CA-II was seen in apical in epithelium of the epididymis in the initial and middle segment. CA-I activity was only founded in ductus deferentis and ampulla ductus deferentis. No or a weak reaction for CA-I, CA-II and CA-III were seen in the three accessory reproductive glands. Specific immunostaining for CA-II and CA-I could be observed in the organ, suggesting the bicarbonate in bovine semen to derive primarily from the genital tract and not accessory reproductive organs. CA-III-positive peritubular cells in interstitial tissue were also stained for alpha smooth muscle actin, and were very similar to contractile myofibroblast cells (Wrobel et al., 1979).

Effects of anordiol, an antiestrogen, on the reproductive organs of the male rat.

Anordiol, an antiestrogen, was administered at doses of 2.5 and 10 mg/kg day-1 for 30 and 60 days to adult male rats. A significant decrease in serum LH, FSH, and testosterone levels occurred in the treated animals as compared to controls. Serum testosterone and LH levels decreased to about 15-35% of control value on day 15 of treatment and reached undetected levels thereafter. Serum FSH level also decreased to about 25-50% of control value by day 15 of treatment and remained at this level. Significant decrease in the weights of the tests and accessory reproductive organs occurred following anordiol treatment, while the body weights remained at pretreatment level. Histological examination of the tests revealed significant decrease in the diameter of the seminiferous tubules and in the number of germ cell elements. Spermatogenesis was arrested at the secondary spermatocyte stage. Leydig cells appeared atrophic and contained pyknotic nuclei. The epididymis, prostate, and seminal vesicle showed degenerative changes. The secretory activity of the glandular epithelium of the prostate gland appeared to be markedly reduced. In conclusion, anordiol acts by blocking gonadotropin production and/or release by the pituitary; thereby testosterone production by Leydig cells is not stimulated, causing spermatogenesis arrest.

Physiological and pharmacological activities of simple alkyl- and arylpyrazines

This review describes the physiological and pharmacological effect of simple alkyl- and arylpyrazines. The description is made in the following order 1) the presence of 2,5-dimethylpyrazine in the urine samples of human being and rodents 2) the effects of tetramethyl- and tetraethylpyrazines on the vascular smooth muscles 3) the activity of alkyl- and arylpyrazines on the aggregation of arachidonic acid- and collagen-induced platelets 4) the effects of 2,5-dimethylpyrazine on the reproductive and the accessory reproductive organs of female and male rats 5) the effects of 2,5-dimethylpyrazine on the plasma and on the contents of polyamines and fructose in the accessory reproductive organs of rats.

Influence of photoinhibition on GABA and glutamic acid levels, and on glutamate decarboxylase activity in the testis and epididymis of the golden hamster

Gamma-aminobutyric acid (GABA) is found in the gonads and accessory reproductive organs, and a direct effect on steroidogenesis and sperm viability and motility has been described. The golden (Syrian) hamster is a seasonal breeder, and a pattern of regression-recrudescence in their reproductive organs is observed when adult animals are exposed to less than 12.5 h daylight for an extended period of time. The purpose of this study was to investigate: (1) the presence of GABA in the testis and epididymis of golden hamsters undergoing regression and spontaneous recrudescence; (2) glutamic acid levels and glutamate decarboxylase (GAD) activity in both tissues, and (3) testicular and epididymal testosterone, dihydrotestosterone and 5 alpha-androstane-3 alpha, 17 beta-diol concentrations. Adult golden hamsters were exposed to long (LP 14L:10D) or short (SP 6L:18D) photoperiods for 9, 12, 14, 16, 18 or 22 weeks. When animals were exposed to SP for 14-16 weeks, the testis and epididymis reached maximal involution. Testicular and epididymal androgen levels showed a marked decrease (p < 0.05) during the regression period, and after 18-22 weeks, values began to recover. Between 12 and 18 weeks in SP, the testicular and epididymal content of GABA and glutamic acid was reduced significantly. The concentration of GABA in both tissues showed a sharp rise (p < 0.05), while the concentration of glutamic acid diminished during the period of maximal involution (p < 0.05). In the testis, GAD activity was increased (p < 0.001) after 14 weeks in SP, with no change in the epididymis. In conclusion, glutamic acid via GAD activity could be the main source of GABA in the testis, but not in the epididymis. Furthermore, the presence of GABA in testicular cells and its subsequent photoperiodic variations might act as an important autocrine and/or paracrine modulatory signal in gonadal processes.