Decreased sperm number and motile activity on the F1 offspring maternally exposed to butyl p-hydroxybenzoic acid (butyl paraben)

Butyl p-hydroxybenzoic acid (butyl paraben, BP) is widely used as a preservative in food and cosmetic products. Routledge et al showed that BP is weakly estrogenic in both in vitro and in vivo (rat uterotrophic) analyses. We investigated whether maternal exposures to BP during gestation and lactation periods affected the development of the reproductive organs of the F1 offspring. Pregnant Sprague-Dawley rats were injected subcutaneously with 100 or 200 mg/kg of BP from gestation day (GD) 6 to postnatal day (PND) 20. In the group exposed to 200 mg/kg of BP, the proportion of pups born alive and the proportion of pups surviving to weaning were decreased. The body weights of female offspring were significantly decreased at PND 49. The weights of testes, seminal vesicles and prostate glands were significantly decreased in rats exposed to 100 mg/kg of BP on PND 49. In contrast, the weights of female reproductive organs were not affected by BP. The sperm count and the sperm motile activity in the epididymis were significantly decreased at doses of 100 and 200 mg/kg of BP. In accordance with the sperm count in the epididymis, the number of round spermatids and elongated spermatids in the seminiferous tubule (stage VII) were significantly decreased by BP. Testicular expression of estrogen receptor (ER)-alpha and ER-beta mRNA was significantly increased in 200 mg/kg of BP treated group at PND 90. Taken together, these results indicated that maternal exposure of BP might have adverse effects on the F1 male offspring.     

Treatment of fibroadenomatous hyperplasia in cats with aglépristone

Fibroadenomatous hyperplasia (FAH) is characterized by a rapid proliferation of mammary stroma and duct epithelium of 1 or more glands and predominantly affects younger female cats. Endogenous progesterone and exogenous progestogens play an important role in the genesis of FAH. The presence of progesterone receptors in fibroadenomatous tissue allows for targeted endocrine therapy with progesterone receptor blockers. We report on 22 young cats with FAH, none of which had responded to the withdrawal of progestogens or ovariectomy. The common signs were tachycardia (11 cats); skin ulceration, painful mammary glands, or both (16 cats); lethargy (8 cats); and anorexia (4 cats). The cats were treated with subcutaneous injections of the progesterone receptor blocker aglépristone on 1 (7 cats, 20 mg/kg) or 2 consecutive days (15 cats, 10 mg/kg/d) once weekly. All but 1 cat responded with a complete and lasting remission of signs after 1-4 weeks of treatment. Two cats had a short-term skin irritation at the site of the aglépristone injection. Two pregnant cats with FAH aborted after treatment with aglépristone and subsequently developed endometritis. In conclusion, the results of this study demonstrate that FAH in cats can be treated successfully with the progesterone receptor blocker aglépristone.     

Neonatal estradiol exposure alters uterine morphology and endometrial transcriptional activity in prepubertal gilts

Porcine endometrial development between birth (postnatal day = PND 0) and PND 56 involves differentiation of glandular epithelium (GE) from luminal epithelium (LE) and estrogen receptor-alpha (ER) expression. Juvenile ER architecture evolves after birth, as stroma and nascent GE first express ER. Mature ER architecture is evident after PND 30, when stroma, GE and LE are ER-positive. When administered during discrete periods between PND 0 and 56, effects of estradiol-17beta valerate (EV) on the neonatal porcine uterus relate to endometrial ER architecture. Transient EV exposure from birth reduces embryo survival in pregnant adult gilts. Effects of EV, administered as juvenile endometrial ER architecture develops (P1, PND 0-13), or after mature ER architecture is established (P2, PND 42-55), were evaluated in uteri from gilts treated with corn oil or EV in P1 or P2 and hysterectomized on PND 100 without additional steroids (NSt), on PND 102 after EV on PND100-101 (EV2), or on PND 117 after EV2 followed by progesterone on PND 102-116 (EP). Neonatal EV reduced uterine weight (P < 0.02), size (P < 0.01), luminal protein content (P < 0.07), and percent incorporation of 3H-leucine into nondialyzable endometrial products in vitro (P < 0.01). Group (NSt, EV2, EP) -specific treatment effects detected for endometrial ER, progesterone receptor, uteroferrin, and/or retinol binding protein mRNA levels were frequently related to period (P1,P2). Results support the idea that estrogen-sensitive postnatal organizational events, including those defined, in part, by endometrial ER architecture, are likely components of genetic and epigenetic programs governing uterine morphogenesis and ontogeny of endometrial function in the pig.     

Inhibition of prolactin in the last trimester of gestation decreases mammary gland development in gilts

Prolactin is required for mammary development in various species but its possible role for mammogenesis in pigs is not known. The goal of the present study was therefore to determine the effect of prolactin inhibition by bromocriptine during the last third of gestation on mammary gland development in gilts. Twenty-eight primigravid gilts were assigned as controls (n = 15) or received 10 mg of bromocriptine orally thrice daily (n = 13) from d 70 to 110 of gestation. Jugular blood samples were collected on d 70 of gestation and every 8 d thereafter and were assayed for prolactin, IGF-I, estradiol, and progesterone. Gilts were slaughtered on d 110 of gestation and fetuses were counted and weighed. One row of mammary glands was used for dissection of parenchymal and extraparenchymal tissues and for determination of DNA, RNA, dry matter, protein, and fat contents. Tissue from the other row was used for measures of prolactin receptor number and affinity. Concentrations of prolactin were drastically reduced throughout the bromocriptine treatment period (P < .001), whereas there was no overall treatment effect on progesterone and IGF-I levels (P > .10). Total weight and extraparenchymal tissue weight of the mammary glands were unaffected by treatment (P > or = .1), but weight of parenchymal tissue, total DNA, and total RNA decreased (P < .01) with bromocriptine treatment. Percentages of fat and dry matter in parenchymal tissue increased with bromocriptine treatment (P < .01) and the percentage of protein decreased (P < .01). Number of prolactin receptors in parenchymal tissue decreased with bromocriptine treatment (P < .001) and receptor affinity increased (P < .001). Average fetal weight was lower in gilts receiving bromocriptine than in control gilts (P = .05), but fetal number did not differ (P > .1). These results clearly demonstrate that prolactin is essential for normal mammary gland development and can affect fetal growth during the last third of gestation in gilts.     

Histomorphometric and Progesterone Receptor Immunohistochemical Analysis in the Oviduct of Newly Hatched Chicks Treated with Follicle-Stimulating Hormone during Embryonic Development

In this study we evaluated the histomorphology and ultra-structure of the oviduct of newly hatched chicks, as well as the immunohistochemical expression of progesterone receptor (PR) in this tissue after follicle-stimulating hormone (FSH) treatment on days 13, 15 and 17 of embryonic development. Results indicated a marked difference in the histology of the oviduct of newly hatched chicks treated with FSH. Magnum mucosa from these animals presented a pseudostratified epithelium with evaginations from the lumen into the epithelium and from the latter into the stroma beneath where tubular glands are formed. In contrast magnum mucosa from control animals presented columnar epithelium with no evaginations. In magnum epithelium FSH also induced the formation of cilia and microvilli projections into the lumen as well as an increase in the wall and lumen areas and in the density of nuclei per unit-area. PR immunoreactivity was only observed in the oviduct of FSH treated animals. PR was located in the nucleus of epithelial luminal cells, mucosal stromal cells and smooth muscle cells. These findings suggest that FSH induces an adequate hormonal milieu for the cytodifferentiation and PR gene expression in the chick oviduct.     

A study of the immunohistochemical localization of the progesterone and oestrogen receptors in the magnum of the immature ostrich,Struthio camelus

The immunolocalization of the progesterone (PR) and oestrogen receptors (OR), in the magnum of the immature ostrich, was investigated during periods of ovarian activity and inactivity. In the immature ostrich, with an active ovary, numerous well-developed tubular glands were present in the lamina propria. Significantly, PR immunostaining was strong in the surface epithelium and tubular glands of these birds. In contrast, weak staining for the PR was observed in the surface epithelium of birds with inactive ovaries. Tubular gland formation, in these birds, was indicated by bud-like invaginations of the surface epithelium. Oestrogen receptor immunoreactivity was negligible in both birds with active and inactive ovaries. These findings suggest that steroid hormones, produced by the active ovary of the immature ostrich, influence the differentiation of the magnum. Furthermore, the action of these steroid hormones appears to be mediated through the PR.     

Molecular studies on oestrogen α and progesterone receptors and histomorphometric analysis of canine uteri following aglepristone treatment

Aglepristone, a competitive progesterone antagonist, is successfully used in various progesterone-dependent conditions. This study investigated uterine histomorphometric analysis, and expressions of the oestrogen α receptor (ERα) and progesterone receptor (PR) in uteri of bitches following the single dose of aglepristone treatment. Twelve client-owned healthy diestrous bitches were used in the study. The single dose of aglepristone (Alizine^®, 10 mg/kg) was injected subcutaneously 5 days before ovariohysterectomy in the treatment group (n = 6); bitches without treatment served as a control group (n = 6). Uteri were collected for histomorphometric analysis, ERα and PR gene, and protein expressions studies. The mRNA expressions of ERα and PR were determined by RT-qPCR. Immunohistochemical analysis was used to evaluate the ERα and PR protein expressions using an H-score in five parts of the uterus. The results demonstrated glandular epithelium height significantly decreased (p < .05) and ERα mRNA increased (p < .01) in treated dogs. Of the treated bitches, lower expression levels of ERα were observed in the luminal epithelium, crypt and glandular epithelium, with higher expression in the endometrial stroma and myometrium (p < .05); however, PR expression decreased in the luminal epithelium, crypt and glandular epithelium (p < .01). In conclusion, reduction of the uterine glandular epithelium and ERα mRNA upregulation together with changes in ERα and PR expressions were observed in the treated bitches. However, changes in uterine ERα and PR expressions in the treated bitches depended on tissue layers. The treatment had no effect on serum oestradiol and progesterone levels.     

Sex differences in oestrogen receptor levels in adrenal glands of sheep during the breeding season

The concentrations of the oestrogen receptor (ER), and the mRNA levels of ER, progesterone receptor (PR) and insulin-like growth factor I (IGF-I) were characterised in adrenal glands and uterine tissue of adult Corriedale sheep during the breeding season. The sheep were of different sex and gonadal status. Ewes had higher levels of cytosolic ER in the adrenals than the rams (mean±S.E.M.: 7.3±2.0 fmol/mg protein and 2.5±1.0 fmol/mg protein, respectively; P=0.0091) and gonadectomy increased ER (mean±S.E.M.: 2.9±1.2 fmol/mg protein and 8.6±2.3 fmol/mg protein, intact and gonadectomised sheep, respectively; P=0.0071). No differences could be observed in mRNA levels for ER and IGF-I in the adrenal glands of all of the sheep. PR mRNA levels were reduced in ovariectomised ewes and enhanced in castrated rams (sex×gonadal status: P=0.009). PR mRNA levels tended to be higher in ewes in the follicular phase than in ovariectomised ewes and intact rams (P<0.1). All of the animals had positive nuclear staining for ER in the adrenal cortex, but no differences were observed between the groups. In this study, we demonstrated the existence of ER in the adrenal gland of sheep and found varying sensitivity to oestrogens as the ER levels differed among sex and gonadal status. These findings indicate that oestrogens most likely affect steroidogenesis directly at the adrenal cortex and suggest that oestrogens are partly responsible for the sex differences in cortisol secretion in sheep.     

Specific window of prolactin inhibition in late gestation decreases mammary parenchymal tissue development in gilts

Prolactin is required from d 70 to 110 of gestation for normal mammary development of gilts. The goal of the present study was to determine the effect of inhibiting prolactin with bromocriptine during specific time windows during the second half of gestation on mammary gland development in gilts. Crossbred primigravid gilts were assigned as controls (n = 12) or received 10 mg of bromocriptine orally three times daily from d 50 to 69 (BR50, n = 12), d 70 to 89 (BR70, n = 12), or d 90 to 109 (BR90, n = 12) of gestation. Jugular blood samples were collected on d 50, 70, 90, and 109 of gestation and assayed for prolactin and estradiol. Gilts were slaughtered on d 109 of gestation and fetuses were counted and weighed. One row of mammary glands was used for dissection of parenchymal and extraparenchymal tissues, and for biochemical analyses. Tissue from the other row was used for measures of prolactin receptor number and affinity. Concentrations of prolactin were decreased markedly (P < 0.001) at the end of each bromocriptine treatment period compared with controls, but there was no overall treatment effect (P > 0.1) on estradiol concentrations. Extraparenchymal tissue weight of the mammary glands was unaffected by treatments (P > 0.1), but weight of parenchymal tissue, total DNA, and total RNA were lower (P < 0.01) in BR90 than control gilts. The percentage of DM in parenchymal tissue was unaffected by treatments (P > 0.1), but percentage of fat was higher and percentage of protein lower (P < 0.01) in BR90 gilts compared with controls. Cell size, as estimated by the protein:DNA ratio, also was lower (P < 0.01) in the BR90 group. Number and affinity of prolactin receptors in parenchymal tissue were not significantly altered by treatments. In conclusion, there is a specific time period in the second half of gestation, from 90 to 109 d, during which prolactin is essential for normal mammary parenchymal tissue development.     

Cell‐specific Distribution of Progesterone Receptors in the Bovine Ovary

This study describes the localization of progesterone receptors (PR) in the bovine ovary. Ovaries were obtained from 11 non‐pregnant and two pregnant cows. Progesterone receptors were visualized by immunohistochemistry on paraffin sections. Nuclear staining for PR was observed in cells of the follicles, corpora lutea, theca layers, surface epithelium, tunica albuginea, and in superficial and deep stroma cells. No staining was noticed in apoptotic bodies of atretic follicles. Expression of PR in follicle cells indicates an intrafollicular role of progesterone. The higher expression in thecal cells compared with follicle cells indicates that thecal cells mediate some effects of progesterone on the follicular development. Superficial stroma cells showing high expression might have a similar influence on primordial and primary follicles. In general, luteal cells had a lower expression than follicle cells, which may be explained by the down‐regulatory effect of locally produced progesterone. The lower expression in luteal cells during pregnancy can be due to the longer life span of this corpus luteum and concomitant degeneration of its PR. The high and rather constant expression of PR in cells of the surface epithelium remains to be elucidated.     

Erratum

This study describes the localization of progesterone receptors (PR) in the bovine ovary. Ovaries were obtained from 11 non‐pregnant and two pregnant cows. Progesterone receptors were visualized by immunohistochemistry on paraffin sections. Nuclear staining for PR was observed in cells of the follicles, corpora lutea, theca layers, surface epithelium, tunica albuginea, and in superficial and deep stroma cells. No staining was noticed in apoptotic bodies of atretic follicles. Expression of PR in follicle cells indicates an intrafollicular role of progesterone. The higher expression in thecal cells compared with follicle cells indicates that thecal cells mediate some effects of progesterone on the follicular development. Superficial stroma cells showing high expression might have a similar influence on primordial and primary follicles. In general, luteal cells had a lower expression than follicle cells, which may be explained by the down‐regulatory effect of locally produced progesterone. The lower expression in luteal cells during pregnancy can be due to the longer life span of this corpus luteum and concomitant degeneration of its PR. The high and rather constant expression of PR in cells of the surface epithelium remains to be elucidated.     

Changes in oestrogen,progesterone and epidermal growth factor receptor concentrations and affinities during the oestrous cycle in the normal mammary gland and uterus of dogs

Changes in the concentrations and affinities of receptors for oestrogen (ER), progesterone (PR) and epidermal growth factor (EGF-R) were studied in mammary glands of healthy bitches with regard to age, the location in the mammary chain and the stage of the oestrous cycle. Uterus was used as the reference tissue for the evaluation of steroid receptors. Mammary and uterine samples from 7 healthy bitches were taken at five stages of the oestrous cycle in such a way that all the locations in the mammary chain were represented at each stage of the cycle (10 samples/dog). ER, PR and EGF-R were detected by biochemical assays using increasing concentrations of tritiated (steroids) or iodinated (EGF) ligands. A significant direct correlation was found between the ER and PR concentrations for mammary and uterine samples. No significant correlation was found between the steroid receptors and EGF-R concentrations. Mammary ER concentrations were significantly higher in bitches of 5 years of age or older than in younger ones; in posterior glands (4th and 5th pairs) than in anterior glands; and in the mid-luteal phase. Mammary PR did not vary significantly with age or location but was significantly lower in the early luteal phase than in other phases. A similar decrease in PR concentrations was observed in the uterus during the early luteal phase and uterine ER and PR concentrations were very low in the mid-luteal phase. Mammary EGF-R were not significantly higher in the early or mid-luteal phase than in pro-oestrus or anoestrus.The differences observed between the uterine and mammary steroid receptor concentrations during the oestrous cycle could be due to different mechanisms for regulating steroid receptor expression in the two tissues. Mammary EGF-R concentrations may be linked, as in other species, to cellular proliferation and/or to the serum progesterone concentrations.Abbreviations BSA bovine serum albumin - EGF epidermal growth factor - EGF-R receptor for epidermal growth factor - ER oestrogen receptor - K _d dissociation constant - LH luteinizing hormone - p probability of error - PR progesterone receptor     

Evaluation of the estrogenic effects of dietary perinatal Trifolium pratense

This study was designed to investigate the potential estrogenic effects of perinatal dietary phytoestrogens on the rat uterus. Pregnant rats were divided to three groups provided the following diets: (1) rat chow, (2) rat chow with 7.5% Trifolium (T.) pratense, or (3) rat chow supplemented with 17β-estradiol (0.5 mg/kg). The dams in each group were kept on the same diet during pregnancy and lactation. Female offspring were euthanized on day 21 at which time body and organ weights were recorded and tissue samples were taken for histology. Immunohistochemistry was performed to detect estrogen receptor alpha (ERα) and progesterone receptor (PR) levels. Our results revealed estrogen-like biological effects of perinatal T. pratense exposure. Relative uterus and ovary weights in the experimental groups were increased compared to control. The number of uterine glands and luminal epithelium heights were also increased. However, there were no statistically significant changes detected in the immunostaining intensity of ERα and PR between the groups.     

Immunohistochemical Detection of Receptors for Oestrogen and Progesterone in Endometrial Glands and Stroma during the Oestrous Cycle in Nelore (Bos taurus indicus) Cows

The aim of the present study was to monitor endometrial distribution and concentrations of oestrogen receptors α (ERα) and progesterone receptors (PR) by immunohistochemistry in Nelore cows (Bos taurus indicus) during the oestrous cycle. Blood samples were collected for progesterone measurement and endometrial samples were taken from the uterine horn contra lateral to the corpus luteum in 16 cows at days 0 (ovulation), 5, 9, 13 and 19 of the oestrous cycle. Immunostaining evaluation for ERα and PR in the glandular epithelium and uterine stroma was performed by two methods: positive nuclei counting and staining intensity of the nuclei. Specific positive staining reactions for both receptors were limited to cell nuclei and they were not identified in the cytoplasm. The proportion of ERα positive nuclei had a temporal variation throughout the oestrous cycle in both cell types evaluated and was higher in uterine stroma than the glandular epithelium (p < 0.05). The greatest proportion of ERα stained nuclei was observed at oestrus and during the initial and mid luteal phase (days 5, 9 and 13) (p < 0.05) in the glandular epithelium and at days 0, 5 and 9 in the uterine stroma (p < 0.01). The proportion of PR positive nuclei remained constant throughout the entire oestrous cycle for both cell types evaluated (p > 0.05). A higher proportion of PR positive nuclei was measured in the uterine stroma compared with the glandular epithelium (p < 0.05). Intensity of staining for ERα and PR varied throughout the oestrous cycle (p < 0.01). There was a higher staining intensity at days 0 and 5 in the stroma for ERα (p < 0.01) and PR (p < 0.01) and in the glandular epithelium at days 0, 5, 9 and 13 for ERα (p < 0.01) and at days 0, 5 and 9 for PR (p < 0.01) when compared with the other evaluated days. These data demonstrate that ERα and PR expression varied throughout the oestrous cycle in Nelore cows, in general with highest concentrations at oestrus and the lowest during the luteal phase. This is similar to patterns observed in Bos taurus taurus.     

The effect of pregnancy on visceral growth and energy use in beef heifers

Beef heifers (24 mo; 378 +/- 32 kg of BW; 22 pregnant, PR; 17 nonpregnant, NP) were grouped in common pens and fed corn silage- and hay-based diets formulated to provide an ADG of 0.45 kg in NP heifers. Both PR and NP heifers were slaughtered on d 40, 120, 200, and 270 of the study. Intestinal and hepatic tissues were analyzed for protein, DNA, RNA (mg/g of fresh tissue), and in vitro oxygen use. Jejunal samples were analyzed for cellular proliferation via immunohistochemical analysis. For ileum, DNA, which provides an estimate of cell number per unit of tissue, revealed an interaction (P = 0.06) between pregnancy and slaughter day; both PR and NP decreased with time, but NP increased on d 270 (P = 0.09). Cell number in the ileum was reduced at d 200 and 270 in the PR heifers (P < 0.09). Liver protein concentration was less (P = 0.07) in PR than in NP heifers (NP = 291.1 vs. PR = 210.5 +/- 33.9 mg/g). Hepatic protein:DNA ratio was not affected (P > 0.10) by pregnancy or day. Energy use (kcal/d) of duodenum and jejunum, calculated from in vitro oxygen consumption, increased linearly (P < 0.02) with time for both PR and NP. Pregnant and NP ileal energy use increased linearly (P < 0.01), but ileal energy use by PR was less throughout gestation (P = 0.07) than ileal energy use by NP. Cellular proliferation in the crypt region of the jejunum was decreased on d 120 and 200 (P < 0.02). These data indicate that the small intestine and liver of PR heifers may conserve energy expenditure compared with NP heifers. Energy conservation can partially be explained by differences in growth and cell proliferation and by energy use of the liver and small intestine.     

大鼠乳腺局部免疫的调节机制--泌乳期与静止期乳腺肥大细胞分布的比较

应用改良甲苯胺蓝染色法(MTB)观察了大鼠泌乳期和静止期乳腺肥大细胞的分布、形态、数量变化规律．并用阿尔新蓝一番红鉴别染色法(AB-S)进行了细胞化学分型研究。结果：AB-S染色显示，大鼠乳腺只存在黏膜型肥大细胞；无论是泌乳期还是静止期。乳腺肥大细胞大多分布于腺泡间和小叶问结缔组织中。细胞的形态各异，但细胞数量在静止期和泌乳期有显著差异。泌乳期的乳腺肥大细胞数量明显减少(P＜0．01)。乳腺肥大细胞的动态变化，可能与乳腺泌乳期腺泡上皮的生长和静止期腺泡问结缔组织细胞增生等结构变化有关。     

植物甾醇对去卵巢KM小鼠血清生殖激素及乳腺组织孕激素受体的影响

本试验旨在研究植物甾醇对去卵巢小鼠血清激素及乳腺组织中孕激素受体的影响。50只雌性小鼠,随机选40只进行去卵巢手术,恢复7 d。分组如下：正常组、去卵巢对照组、植物甾醇低、中、高3个剂量组（每日灌胃20、80和320 mg/kg植物甾醇）。连续灌胃3周,第22天采集血样和乳腺组织,对小鼠血清中的雌二醇、孕酮和催乳素水平进行检测,对乳腺组织的孕激素受体基因表达进行荧光定量检测。结果表明,去卵巢对照组小鼠雌二醇水平显著低于正常组（P〈0.05）。植物甾醇组小鼠血清雌二醇水平持续升高,其中,高剂量组小鼠血清雌二醇水平与正常组最接近;与去卵巢对照组相比,去卵巢植物甾醇组小鼠血清孕酮水平均有提高,但差异不显著（P〉0.05）;与去卵巢对照组相比,去卵巢植物甾醇组小鼠催乳素水平先下降后上升,但他们之间无显著差异（P〉0.05）。与去卵巢对照组相比,植物甾醇各剂量组小鼠孕激素受体基因相对表达量呈上升趋势,低、中、高剂量组小鼠孕激素受体基因相对表达量分别是去卵巢对照组的1.05、1.13和2.78倍。植物甾醇可改善去卵巢小鼠内源激素不平衡的状况,促进乳腺组织PR基因的表达。