Effects of progesterone treatment during adulthood on consummatory and motivational aspects of sexual behavior in male rats.

In males of a variety of species, administration of progesterone during adulthood has been shown to decrease the expression of consummatory sexual behaviors and androgen receptors. However, it remains to be determined if the progesterone-induced decrease in androgen-receptor signaling and consummatory sexual behaviors correspond with less of a preference for a sexually receptive female relative to another male, a behavioral phenotype indicative of sexual motivation. Consistent with the effects of progesterone reported in males of other species, progesterone-treated rats, relative to vehicle-treated rats, exhibited fewer intromissions and ejaculations. Correspondingly, the weights of the androgen sensitive bulbourethral glands were lighter in progesterone-treated rats. In addition, unlike vehicle-treated rats, progesterone-treated rats did not exhibit a preference for a female rat during the early stages of testing. However, across the entire test, both treatment groups exhibited a preference for a female rat, and consequently, there were no differences between the conditions in overall sexual motivation. Progesterone treatment did not alter activity or anxiety-like behaviors. The results of the current study suggest that the lower levels of androgen-receptor signaling and consummatory sexual behaviors in males following progesterone treatment are associated with a transient deficit in the preference for a female sexual incentive.

Hydrolysable tannin-based diet rich in gallotannins has a minimal impact on pig performance but significantly reduces salivary and bulbourethral gland size.

Tannins have long been considered 'anti-nutritional' factors in monogastric nutrition, shown to reduce feed intake and palatability. However, recent studies revealed that compared with condensed tannins, hydrolysable tannins (HT) appear to have far less impact on growth performance, but may be inhibitory to the total activity of caecal bacteria. This in turn could reduce microbial synthesis of skatole and indole in the hindgut of entire male pigs (EM). Thus, the objective of this study was to determine the impact of a group of dietary HT on growth performance, carcass traits and boar taint compounds of group housed EM. For the study, 36 Swiss Large White boars were assigned within litter to three treatment groups. Boars were offered ad libitum one of three finisher diets supplemented with 0 (C), 15 (T15) or 30 g/kg (T30) of HT from day 105 to 165 of age. Growth performance, carcass characteristics, boar taint compounds in the adipose tissue and cytochrome P450 (CYP) isoenzymes CYP2E1, CYP1A2 and CYP2A19 gene expression in the liver was assessed. Compared with C, feed efficiency but not daily gain and daily feed intake was lower (P<0.05) in T15 and T30 boars. Except for the percentage carcass weight loss during cooling, which tended (P<0.10) to be greater in T30 than C and T15, carcass characteristics were not affected by the diets. In line with the numerically lower androstenone level, bulbourethral and salivary glands of T30 boars were lighter (P<0.05) than of T15 with intermediate values for C. Indole level was lower (P<0.05) in the adipose tissue of T30 than C pigs with intermediate levels in T15. Skatole levels tended (P<0.10) to be lower in T30 and C than T15 pigs. Hepatic gene expression of CYP isoenzymes did not differ between-treatment groups, but was negatively correlated (P<0.05) with androstenone (CYP2E1 and CYP1A2), skatole (CYP2E1, CYP2A) and indole (CYP2A) level. In line with the numerically highest androstenone and skatole concentrations, boar taint odour but not flavour was detected by the panellists in loins from T15 compared with loins from C and T30 boars. These results provide evidence that HT affected metabolism of indolic compounds and androstenone and that they affected the development of accessory sex glands. However, the effects were too small to be detected by sensory evaluation.

Regucalcin Expression as a Diagnostic Tool for the Illicit Use of Steroids in Veal Calves.

It has been previously demonstrated that sex steroid hormone treatment down-regulates regucalcin gene expression in the accessory sex glands and testis of prepubertal and adult male bovines. The aim of this study was to investigate whether low doses of sex steroid hormones combined with other drugs significantly affect regucalcin gene expression in the accessory sex glands and testis of veal calves. The regucalcin expression was down-regulated in the bulbo-urethral glands of estrogen-treated calves, whereas it was up-regulated in the prostate of estrogen-treated calves. Only the testis of androgen-treated calves showed a down-regulation of the regucalcin expression. Thus, the administration of sex steroid hormones, even in low doses and combined with other molecules, could affect regucalcin expression in target organs. Particularly, the specific response in the testis suggests regucalcin expression in this organ as a first molecular biomarker of illicit androgen administration in bovine husbandry.

Effect of long-term testosterone propionate or human chorionic gonadotrophin administration on reproductive glands in adult male rabbits.

The study was aimed to investigate the effect of testosterone propionate (TP) or human chorionic gonadotrophin (hCG) treatment on reproductive glands in sexually mature male rabbits. A total 36 adult male rabbits were randomly distributed to six equal groups. The first control group (CON), the second treated with low-dose TP (TPL), the third treated with high-dose TP (TPH), the fourth treated with low-dose hCG (CGL), the fifth treated with medium-dose hCG (CGM) and sixth treated with high-dose hCG (CGH). At the 16th post-treatment week, the animals were sacrificed, and the testes and accessory sex glands dissected, weighted and stored at -20 °C until assay. Testosterone propionate treatment in both doses resulted in reduction (P < 0.01) in testicular weight and increase (P < 0.01) in weight of vesicular gland, paraprostate and proprostate glands. High-dose TP increased the weight of prostate and bulbouretheral gland (BUG). Testosterone propionate increased total androgen (P < 0.01) with Testosterone (T) predominating in serum, dihydrotestosterone (DHT) predominating in testes and most accessory sex glands. High dose of hCG increased the weight of proprostate and paraprostate glands. Androgen level in serum, testes and accessory sex glands increased (P < 0.01) after hCG treatment.

Application of absolute qPCR as a screening method to detect illicit 17ß-oestradiol administration in male cattle.

It has been previously demonstrated that the progesterone receptor gene is up-regulated in the sex accessory glands of pre-pubertal and adult male bovines after 17ß-oestradiol treatment. In the present study, a qualitative screening method was optimised to detect 17ß-oestradiol treatment using absolute quantification by qPCR of the progesterone receptor gene to determine the amount of gene expression in bulbo-urethral glands. An external standard curve was generated and developed with TaqMan® technology. Based on two in vivo experiments, the decision limit CCα, sensitivity and specificity of this screening method were established. Trial 1 consisted of 32 Friesian veal calves divided into two groups: group A (n = 12), consisting of animals treated with four doses of 17ß-oestradiol (5 mg week(-1) per animal); and group B (n = 20), consisting of control animals. Trial 2 was performed on 26 Charolaise beef cattle that either received five doses of 17ß-oestradiol (group C; 20 mg week(-1) per animal; n = 6) or remained untreated (group D; n = 20). Further, progesterone receptor gene expression was evaluated in beef and veal calves for human consumption. A specific CCα on 20 Piedmontese control beef cattle was calculated to include these animals in a field investigation. Five out of 190 beef cattle and 26 out of 177 calves tested expressed the progesterone receptor gene above their respective CCα and they were classified as being suspected of 17ß-oestradiol treatment. Additionally, 58% of veal calves that tested suspect via qPCR exhibited histological lesions of the bulbo-urethral gland tissue, which are typical of oestrogen administration and are consistent with hyperplasia and metaplasia of the glandular epithelium.

Illegal treatment of barrows with nandrolone ester: effect on growth, histology and residue levels in urine and hair.

The effect of 17ß-19-nortestosterone (17ßNT) treatment of barrows on residue levels and growth was evaluated. Five barrows were treated three times during the fattening period with 17ßNT phenylpropionate (Nandrosol, nandrolone phenylpropionate 50 mg/ml,1 mg/kg body weight). Another five barrows were untreated and five boars (untreated) were kept as positive control. Boars and treated barrows showed a 13 and 9% improvement in growth compared to untreated barrows, with mean final body weights of 121.6, 117.8 and 109.0 kg, respectively. The bulbourethral glands of the treated barrows were three times heavier than untreated barrows. The histology of the prostate and bulbourethral gland of the treated barrows was comparable to the boars, whereas the control barrows showed atrophic glands. Levels of 17ßNT ester in hair from treated barrows were high, whereas boars and untreated barrows did not show levels above LLQ. It is concluded that analysis of hair can detect illegal treatment with 17ßNT ester in barrows. The size of the bulbourethral gland can also be used for screening in the slaughterhouse.

Effects of early vaccination with Improvac(®) on the development and function of reproductive organs of male pigs.

Gonadotropin-releasing hormone (GnRH) vaccine (Improvac(®)) is effective at diminishing boar taint by interfering with testis function. Early pre-pubertal vaccination at 10 and 14 weeks-of-age could be desirable if sufficient and sustained effects could be achieved. Crossbred male pigs (n=24) were randomly assigned to three groups each with eight individuals: an unvaccinated control group, one group vaccinated with Improvac(®) early at ages 10 and 14 weeks, and a third group vaccinated with Improvac at the standard ages of 16 and 20 weeks. The average age at slaughter was 25 weeks. At slaughter, reductions in testes weight and bulbourethral gland length of vaccinated pigs compared with controls were observed (P<0.001), accompanied by lowered testosterone concentrations in peripheral blood (P<0.001). The diameter of tubuli seminiferi was affected; being 18% smaller in standard and 38% smaller in early vaccinated males, compared with controls (P<0.01). Leydig cells in vaccinated pigs became pycnotic, and their number decreased in early vaccinated pigs. Spermatogenesis was disrupted, evidenced by spermatocyte loss among standard vaccinated pigs to severe spermatogenic arrest among early vaccinated pigs. This histological picture was reflected in the absence of epididymal spermatozoa in 5 of 8 early vaccinated pigs and a dramatic reduction in the remaining 3 early vaccinated pigs. Among standard vaccinated pigs, 5% of the spermatozoa were morphologically normal (>70% in controls, P<0.01). Early vaccination caused a more severe disruption of testicular structure and function than standard vaccination, thus providing an alternative for immunocastration of male pigs.

Effects of a diet containing fusarium toxins on the fertility of gilts and on bulbourethral gland weight in barrows.

Nine gilts weighing 80 kg at the beginning of the trial were fed a mycotoxin contaminated diet containing 2 mg deoxynivalenol (DON) and 0.4 mg zearalenone (ZON) per kg (Diet M). Their daily weight gain until 103 kg BW was reduced in comparison to the nine control animals fed an uncontaminated diet (Diet C) (763 vs. 912 g; p = 0.02). There was no treatment effect on the age at first observed oestrus. Seven and eight gilts receiving Diet M and C, respectively, became pregnant after being mated once or being again mated three weeks later. The examination of the uteri of gilts slaughtered 35-61 days after mating showed that the exposure to DON and ZON had no effect on the number of foetuses per gilt (p = 0.54), but increased their growth rate (p = 0.003). Thus, low dietary DON and ZON levels had no negative effects on the reproductive parameters examined. The hypothesis that the bulbourethral gland weight of barrows can be used for the bioassay of low dietary ZON levels was rejected since feeding Diet M from 80-103 kg BW did not increase the weight of that accessory sex gland (p = 0.51).

Morphology and morphometry of the reproductive organs in prepubertal and postpubertal male pigs exposed to di(2-ethylhexyl) phthalate before puberty: Precocious development of bulbourethral glands.

In a split-litter design experiment, male piglets were exposed orally three times weekly to 300 mg/kg of di(2-ethylhexyl) phthalate (DEHP) or placebo between three and seven weeks of age. The effects on the reproductive organs were examined immediately after the exposure at seven weeks of age in one sub-group, and postpuberally at nine months of age in the other. Morphological features of testes were unaffected at either age group; there were no differences (p>0.05) between the treatments in number of Sertoli cells (as identified by immunostaining with GATA-4 antibodies), percent area of Leydig cells (as detected by 3beta-hydroxysteroid dehydrogenase histochemistry), or incidence of germinal epithelial lesions (histopathology of H&E-stained (hematoxylin and eosin) sections). Three of the seven DEHP-treated animals in seven-week-old group had bulbourethral glands at a stage of maturation far more advanced than that of controls. While there were no obvious differences in the cellular composition between the treatment groups in nine-month-old animals, the bulbourethral glands were heavier (p<0.05) in DEHP-treated boars. Collectively, these features indicate that adolescent exposure to DEHP induces precocious maturity of bulbourethral glands in pigs with persistent effects lasting into adulthood.

Pathology of the testicle and sex accessory glands following the administration of boldenone and boldione as growth promoters in veal calves.

Boldenone and its precursor Boldione are illegally used for anabolic purposes in humans, horses and cattle. To develop more effective policies and programs to maximize food security, Italian Public Health Services investigate all indicators capable of assisting the recognition of treated animals, and prioritize research and the formulation of action strategies for the promotion of healthy eating. Thus, an experimental administration of boldenone and boldione at anabolic dosages in veal calves was carried out to evaluate the changes in target organs by qualitative and semi-quantitative morphological analysis. The lesions resembled the effects already observed after the administration of androgen hormones to cattle. Main findings were represented by prostate hypersecretion, increased rate of apoptotic cells and decreased rate of Ki67 positive cells in the germ cell line of treated animals, particularly in boldione group and finally some new features like hypertrophy of the prostate urothelial cells.

Potential estrogenic and antiandrogenic effects of permethrin in rats.

Many environmental chemicals including pesticides have been reported to possess hormonal activities, and thus are classified as endocrine disruptors. Permethrin, a synthetic pyrethroid insecticide, is used worldwide, which provides potential environmental exposure. However, relatively few studies have reported on hormonal activities, particularly estrogenic and androgenic activities of permethrin, and the results of these studies are in some respects contradictory. Therefore, this study investigated the potential estrogenic and androgenic activities of permethrin in vitro and in vivo. We conducted an uterine Calbindin-D9k (CaBP-9k) gene expression assay and an uterotrophic assay for estrogenic activity, and a Hershberger assay for androgenic activity. The CaBP-9k gene, one of the intracellular calcium binding proteins, is estrogen-responsive in the uterus. The rat uterotrophic and Hershberger assays are generally used as in vivo short-term screening assays for detecting the estrogenic and androgenic activities of chemicals, although these assays are still being validated by the Organization for Economic Cooperation and Development (OECD). Northern blot analysis showed the induction of uterine CaBP-9k mRNA level in response to permethrin as well as co-administration of permethrin with E2. In the uterotrophic assay using 18-day-old female rats, subcutaneous treatments with permethrin (10 to 800 mg/kg) for three days increased relative uterine wet weights, and E2-induced uterine weights. These effects were statistically significant at 800 and 200 mg/kg, respectively. Moreover, permethrin-induced uterine weights were inhibited by the co-administration of ICI 182,780, an antiestrogen. In the Hershberger assay, the administration of permethrin orally to testosterone propionate-treated castrated male rats led to statistically significant reductions in androgen-dependent sex accessory tissue (ventral prostate, seminal vesicles, levator ani and bulbocavernosus muscles, Cowper's gland and glans penis) weights at all doses tested (10, 50 and 100 mg/kg). These results suggest that permethrin might have estrogen-like effects on female rats, but antiandrogen-like effects on males.

Effects of transforming growth factor beta-1 and all-trans-retinoic acid on androgen-induced development of neonatal mouse bulbourethral glands in vitro.

Effects of transforming growth factor beta-1 (TGF-beta1) and all-trans-retinoic acid (All-trans-RA) on development of bulbourethral glands (BUGs) of neonatal mice were investigated in vitro. BUGs from 0-day-old male mice were cultured for 6 days in serum-free, chemically defined medium containing transferrin and bovine serum albumin, supplemented with 5alpha-dihydrotestosterone (DHT; 10-8 M) and insulin (10 microg/mL) alone or in combination. Prior to culture, BUGs from 0-day-old mice consisted of a simple epithelial rudiment encapsulated by mesenchyme. Epithelial growth and ductal branching occurred in BUGs cultured in medium containing DHT and insulin or DHT alone, but epithelial branching did not occur in BUGs cultured in the presence of insulin alone. Addition of TGF-beta1 at concentrations of > 5 ng/mL (0.2 x 10-9 M) to medium containing both insulin and DHT, inhibited the expected increase in overall size of BUGs, epithelial area and ductal branching in a dose-dependent manner. TGF-beta1 also decreased [3H]-thymidine labelling indices of both epithelium and mesenchyme. TGF-beta1 at 10 ng/mL elicited these inhibitory effects on BUGs cultured in medium containing DHT alone. Addition of All-trans-RA (10-8 to 10-6 M) to the medium containing DHT plus insulin, or DHT alone did not exert significant effects on either overall size of BUGs or epithelial growth and ductal branching. All-trans-RA at 10-6 M decreased the [3H]-thymidine labelling index of mesenchyme of BUGs cultured in medium with DHT plus insulin or DHT alone, but did not decrease the [3H]-thymidine labelling index of epithelium. The present results indicate that TGF-beta1 inhibits androgen-induced epithelial and mesenchymal growth as well as epithelial morphogenesis of BUGs from neonatal mice. Such an inhibitory effect of TGF-beta1 is not mimicked by All-trans-RA at physiological concentrations.

Effect of testosterone and cortisol administration on the reproductive tract of male Antechinus stuartii (Marsupialia).

The life history of Antechinus stuartii, a marsupial, is highly synchronized and culminates in a brief mating period that is followed by complete male mortality. The accessory reproductive tracts of male A. stuartii enlarge in association with testosterone and cortisol hormone concentrations, but this appears to be unrelated to the spermatogenic cycle. The present study examined the effects of testosterone and cortisol on the male reproductive tract. Four groups of adult males from May (when plasma testosterone and cortisol concentrations are low) were given depot injections of testosterone esters or synthetic cortisol in doses that mimic concentrations found in males in the breeding period (August). Males were given either saline, testosterone only, cortisol only, or testosterone plus cortisol. Experimental groups did not differ in the seminiferous tubule morphology. However, the cells from the caudal end of the epididymides of both testosterone groups were considerably hypertrophied compared with males treated with saline or cortisol only. Testosterone treatment significantly increased prostate and bulbourethral gland mass, although addition of cortisol to the testosterone administration diminished this effect. The morphology of the accessory reproductive tract of males treated with either saline or cortisol only was similar to that of untreated males at the same time of year, and the morphology of the accessory reproductive tract of males treated with testosterone plus cortisol was similar to that of untreated males in the breeding season. Like some other marsupials, the spermatogenic cycle in A. stuartii is apparently not correlated with androgen activity, while the accessory reproductive tract is affected by androgens.

Ultrastructure and morphometry of the urethral glands in normal, castrated, and testosterone-treated castrated male mice.

Recent studies of the urethral glands in the male mouse and rat have suggested that they are testosterone-dependent glands that may be potential sites for secretory immunity in the male genital tract. In the present study we describe the ultrastructural features of these glands in normal mice and provide quantitative data on the sizes of the acinar cells and their organelles in sham-, oil-, and testosterone-treated castrated mice. Acinar cells in urethral glands from normal mice contain numerous secretory granules, prominent Golgi complexes, elongated mitochondria, and an abundance of rough endoplasmic reticulum (RER) with large and dilated cisternae, all of which are features characteristic of secretory cells. In some acinar cells the cisternae of the RER were filled with closely packed, unbranched, straight, tubular structures that were oriented parallel to one another, that radiated from aggregates of dense material, or that were randomly arranged. In other acinar cells the cisternae of the RER showed a network of branching and anastomosing vesicular-like structures whose limiting membranes were occasionally seen in continuity with the membranes of the RER. Secretory acini showed large, unbranched tubules in the acinar lumen. When cut at right angles the large tubules exhibited a distinct fuzzy outer coat with fine projections radiating outwards. The ultrastructure of the acinar cells and the presence of tubules in the lumen suggests that they are engaged in secretion of a tubular protein. Morphometric analysis of acinar cells in the urethral glands showed that the mean volumes of nuclei, cytoplasm, secretory granules, vacuoles, and mitochondria were significantly reduced in castrated mice in comparison to either normal or testosterone-treated castrated mice. This confirms earlier observations that the urethral glands are targets of testosterone.

Androgens lower prostaglandin E2 levels in neonatal mouse bulbourethral gland in in vitro cultures.

The neonatal mouse bulbourethral gland (BUG) in vitro culture model is useful to study hormone-induced genitourinary (GU) tract growth and differentiation. Like the prostate, the BUG is a derivative of the urogenital sinus and may have relevance to understanding growth processes involved in normal and pathological GU tract development. Previous studies have reported androgen-induced elevation of prostaglandin E2 (PgE2) levels in mouse GU tract in vivo. PgE2 has been proposed to mediate neonatal GU tract masculinization. In our studies, tissues were obtained from neonatal male mice and cultured in serum-free Dulbecco's Modified Eagle's Medium-Ham's F-12 Medium (1:1) supplemented with varying concentrations of androgen. PgE2 levels were measured by RIA in the medium, and tissue specimens were cultured for 7 days or less. During this period, androgens induced proliferation and glandular morphogenesis in the BUGs. In the absence of androgen, tissue and medium PgE2 levels increased over 7 days. Significant (P < 0.05) PgE2 increases over day 1 control values were observed from days 5-7 in tissues and on day 7 in media. During this same time period, androgen supplementation decreased PgE2 levels. Significant (P < 0.05) PgE2 decreases from day 1 cultures were observed from days 3-7 in tissues and on day 7 in media. PgE2 was decreased significantly (P < 0.05) by androgen compared to control values from days 3-7 in tissues and from days 5-7 in media. On day 7 of culture, PgE2 levels were significantly (P < 0.05) inhibited by androgen in a concentration-dependent fashion in tissues and media. Maximal androgen-induced inhibition of PgE2 levels was 96% and 99% in tissues and media, respectively. Although the addition of indomethacin to control cultures markedly inhibited PgE2 production, BUG morphology was unaffected. In addition, the morphology of androgen-stimulated BUGs does not appear to be affected by the addition of exogenous PgE2. We conclude that although androgens induce development and decrease PgE2 levels, PgE2 does not appear to play a major role in in vitro BUG postnatal growth and morphogenesis. The BUG in vitro culture model may mimic growth and morphogenetic processes occurring in the human GU tract. Further understanding of the role of steroid hormones and PG metabolism may yield additional insight into developmental and proliferative GU tract disorders.

Induction of invasive carcinomas of the seminal vesicles and coagulating glands of F344 rats by administration of N-methylnitrosourea or N-nitrosobis(2-oxopropyl)amine and followed by testosterone propionate with or without high-fat diet.

The potential modifying effects of testosterone propionate (TP) and high-caloric high-fat diet (20% corn oil, HF) on rat accessory sex gland carcinogenesis were investigated. Male F344 rats were treated five times at 4-week intervals with N-methylnitrosourea (MNU) i.v. or N-nitrosobis(2-oxopropyl)amine (BOP) s.c., each injection following 2 weeks pretreatment with dietary ethinyl estradiol. After completion of this carcinogen administration stage, animal groups received subcutaneous implantation of Silastic tubes filled with 40 mg TP with or without HF for 40 weeks. Carcinomas of the seminal vesicles and/or coagulating glands were induced in 5, 39 and 56% of rats given MNU alone, MNU and TP, and MNU and HF plus TP respectively. No equivalent tumors were found in rats given MNU and HF. In the BOP-treated groups, 11% of animals receiving TP but no HF diet demonstrated seminal vesicle carcinomas and 6% of rats receiving TP plus HF diet had coagulating gland carcinoma. Thus while TP exerted a strong enhancing effect on tumor growth in the seminal vesicles and coagulating glands, high caloric HF did not manifest any significant influence.

Induction of invasive carcinomas in the accessory sex organs other than the ventral prostate of rats given 3,2'-dimethyl-4-aminobiphenyl and testosterone propionate.

The promotion effects of testosterone propionate (TP) on prostate carcinogenesis were investigated in F344 rats given the prostatic carcinogen, 3,2'-dimethyl-4-aminobiphenyl (DMAB). One group of animals received s.c. DMAB injections at a dose of 50 mg/kg body weight at 2-week intervals for a total of 10 injections along with s.c. implantations of TP-containing Silastic tubes. A second experimental group of rats was given DMAB at the same dose and intervals but each injection of DMAB was combined with 3 prior consecutive daily 100-mg/kg body weight s.c. injections of TP. After cessation of carcinogen administration, animals in these two groups received TP implants from week 21 to the end of the experiment. All surviving animals were killed at week 56 and accessory sex gland tumor incidences were compared to those in DMAB alone and other appropriate control groups. The groups given TP plus DMAB and subsequent long term administration of TP developed lesions of the dorsolateral prostate, seminal vesicles, and coagulating glands which were all invasive adenocarcinomas. Incidences were 84.2% (16 of 19 rats) and 66.7% (12 of 18 rats), respectively. Macroscopic large tumors were induced in 13 animals among which 8 demonstrated metastasis to the abdominal cavity, liver, or lung. None of the control groups except for the group given TP injections plus DMAB had equivalent tumors. Development of carcinomas of the ventral prostate, which were all of in situ type, were not increased by subsequent treatment with TP. These data thus clearly showed that TP can exert strong enhancing effects on tumor development in the dorsolateral prostate, seminal vesicles, and coagulating glands but not in the ventral prostate.

Clinical, pathological and histopathological findings in lambs implanted with a growth promoting product containing progesterone and oestradiol.

Six live wether lambs and the urogenital tracts of 4 others from 4 separate farms were presented for examination. In each case the wethers had been implanted several weeks previously with a growth promoting product containing progesterone and oestradiol. The affected wethers presented with dorsal retroflexion of a markedly distended bladder which caused persistent straining, bulging of the perineal muscles and rectal prolapse. Enlargement of the bulbo-urethral glands and seminal vesicles, with an increase in the volume and viscosity of the secretion of the bulbourethral glands was evident on gross examination. Histopathological examination of the prostate and bulbourethral glands revealed cystic dilation of the glands and hyperplasia and squamous metaplasia of the glandular epithelium. Desquamated cells and eesinophilic-staining material were present in many of the dilated lumens. In the seminal vesicles, interstitial fibrosis had resulted in the obliteration of large areas of glandular tissue. The enlargement of the prostate and bulbo-urethral glands, and particularly the increased viscosity of the gland secretions, probably hindered urination, resulting in only partial evacuation of the bladder. Attempts to urinate by active contraction of the abdominal muscles may have accounted for the dorsal retroflexion of the distended bladder. The oestrogenic influence of the growth implants is thought to be responsible for these changes.

Inhibitory effects of technical grade methoxychlor on development of neonatal male mouse reproductive organs.

Effects of technical grade methoxychlor (MX), an estrogenic insecticide, and 17 beta-estradiol (E2) were examined on serum testosterone (T) concentrations and growth and histology of neonatal male mouse reproductive organs. Male NIH/Swiss mice received i.p. injections daily from birth to Day 9 with one of the following: 10 micrograms E2 or 0.1 or 1.0 mg MX or sesame oil vehicle. The mice were killed on Day 10. MX or did not affect body weights or mortality. Serum T concentrations in control mice were 0.29 +/- 0.03 ng/ml; T concentrations in males treated with MX or E2 were reduced. The 1.0-mg dose of both MX and E2 significantly decreased DNA contents of the seminal vesicles (SV), bulbourethral glands (BUG), and ventral prostate (VP) compared to controls. In the same animals, DNA contents of testes, epididymides, and efferent ductules were not decreased. The lower dose of MX (0.1 mg) decreased DNA content of only BUG and SV. DNA content of the ductus deferens was not affected by any treatment. E2 and both doses of MX also decreased epithelial morphogenesis in the SV and BUG, and inhibited the onset of mucin production in BUG epithelium and smooth muscle differentiation in the ductus deferens. In summary, our results indicate that technical grade MX, at doses as low as 0.1 mg/day, and E2 inhibit neonatal male reproductive tract development and decrease serum T concentrations.

Anti-androgenic effects of Win 49,596 on development of the neonatal mouse bulbourethral gland in culture.

The effects of the anti-androgen Win 49,596, a steroidal androgen receptor antagonist, on the testosterone (T) dependent development of neonatal mouse bulbourethral glands (BUGs) were examined in vitro. Day of birth (day 0) BUGs were grown in organ culture for 6 days in Dulbecco's modified Eagle's medium:Ham's F-12 media (1:1, volume/volume) with 10% fetal calf serum. Cultures were grown with or without T (10 nM) and various concentrations of Win 49,596 (0-57.6 microns). The DNA content of BUGs grown with T alone increased 4-fold over the culture period and the epithelium grew and branched extensively; without T only minimal growth and epithelial morphogenesis occurred. In the presence of T, Win 49,596 inhibited development in a dose-dependent manner, with an ED50 of 0.8 microM; concentrations at or above 3.6 microM produced maximal inhibition of development. Win 49,596 alone at 14.4 microM did not stimulate BUG growth, demonstrating a lack of agonist activity. BUGs grown for 3 days with Win 49,596 and T, then an additional 6 days with only T, did not resume development. In summary, Win 49,596 produced a dose-dependent suppression of BUG development in vitro, and was not androgenic. Additionally, the inhibitory effects of Win 49,596 persist for at least 6 days following cessation of treatment.