bluuebunny/crossref_metadata_2025_split huggingface dataset:
Influence of thyroid hormone on androgen metabolism in peripuberal rat Sertoli cells
Authors: Panno M L , Beraldi E , Pezzi V , Salerno M , De Luca G , Lanzino M , Le Pera M , Sisci D , Prati M , Palmero S , Bolla E , Fugassa E , Andò S 
Published: December 2008

Abstract
The aim of the present study was to investigate the influence of thyroid hormones on androgen metabolism in Sertoli cells isolated from 3- and 4- week-old rats.
Hypothyroidism was induced by the oral administration of 0·025% methimazole (MMI) from birth until the rats were killed at 3 and 4 weeks of age. Half of the MMI-treated animals were injected i.p. with l-tri-iodothyronine (T3 3 μg/100 g body weight) during the last week before death. Sertoli cells from all groups were initially cultured under basal conditions for the first 24 h and subsequently in the presence of testosterone with or without T3 for an additional 24 h. Hypothyroidism was associated with severe impairment of body as well as testicular growth. Indeed, body and testicular weights were similar in 4-week-old hypothyroid animals to those in 3-week-old control rats.
Testosterone metabolism in Sertoli cells isolated from 3- and 4-week-old hypothyroid rats was mainly expressed by the lowering of 5α-dihydrotestosterone + androstane 3α, 17β–diol and an enhanced formation of 5α-reduced steroids with poor androgenic properties (e.g. 5α–androstane, 3, 17α-dione (androstanedione), 5α–androstan, 3-ol-17-one (androsterone)). Treatment of the same group of animals with T3 in vivo and in vitro did not influence the pattern of 5α–reductase steroids substantially.
The most striking finding in the Sertoli cells of 3-week-old hypothyroid rats was the dramatic enhancement of oestradiol formation which persisted to a lesser extent 1 week later. Oestradiol formation was greatly decreased by the addition of T3 in vivo and in vitro in hypothyroid animals.
These results suggest that T3 might influence androgen metabolism during the functional maturation of Sertoli cells.
Journal of Endocrinology (1994) 140, 349–355
Effects of oestradiol-17β and 5α-dihydrotestosterone on guinea-pig prostate smooth muscle cell proliferation and steroid receptor expression in vitro
Authors: Ricciardelli C , Horsfall D J , Sykes P J , Marshall V R , Tilley W D 
Published: December 2008

Abstract
Smooth muscle cells (SMCs) are the major cellular component of the prostatic stroma. The aim of this study was to examine the effects of oestradiol-17β (OE2) and 5α-dihydrotestosterone (DHT) on the proliferation of guinea-pig prostate SMCs in vitro. OE2 stimulated SMC DNA synthesis at all concentrations examined. At a plating density of 3·0 × 104 cells/cm2, maximal incorporation of [3H]thymidine (136% of control) was observed after 36 h of treatment with 1 nmol OE2/l. At the same plating density, DHT had an inhibitory effect on SMC DNA synthesis, with maximal effects (73% of control) being observed 24 h after treatment with 1 nmol DHT/l. These effects of OE2 and DHT were prevented by co-incubation with specific steroid receptor antagonists. At a threefold lower plating density (1·0 × 104 cells/cm2), the maximal stimulatory and inhibitory effects of OE2 and DHT were delayed by approximately 24 and 12 h respectively. At the lower plating density, a biphasic effect of DHT was observed on DNA synthesis; DHT was both inhibitory and stimulatory. Maximal inhibition (71% of control) and maximal stimulation (168% of control) were observed after 36 and 134 h treatment with DHT respectively. At the lower plating density, longer term treatment of SMC cultures with OE2 and DHT also resulted in an increase in cell number. After 7 days of treatment with OE2 and DHT, cell number increased by 13% and 12% respectively. When OE, and DHT were added in combination, the short-term inhibitory effect of DHT on SMC DNA synthesis was dominant over the stimulatory effect of OE2. Treatment with DHT for 24 h significantly inhibited OE2-induced stimulation of [3H] thymidine incorporation, irrespective of the prior duration of OE2 treatment. At the lower plating density, OE2 also decreased oestrogen receptor (ER) mRNA levels to 38% of control levels after 24 h of treatment. ER mRNA levels remained repressed until 72 h after treatment with OE2, and returned to control values following 96 h of treatment. Both the androgen-induced inhibition and stimulation of DNA synthesis observed following treatment of SMCs with 1 nmol DHT/l were associated with a reduction in androgen receptor (AR) mRNA levels. At an intermediate time (i.e. 48 h after commencement of treatment with DHT) AR mRNA levels were increased more than twofold over control levels. The increase in AR mRNA levels at 48 h after DHT treatment only occurred in cells plated at the lower density, suggesting that this is an essential requirement for the longer term stimulation of prostatic SMC proliferation by DHT.
Journal of Endocrinology (1994) 140, 373–383
Expression of 17β-hydroxysteroid dehydrogenase in the rat ovary during follicular development and luteinization induced with pregnant mare serum gonadotrophin and human chorionic gonadotrophin
Authors: Ghersevich S A , Poutanen M H , Rajaniemi H J , Vihko R K 
Published: December 2008

Abstract
Antibodies against human placental 17β-hydroxysteroid dehydrogenase (17-HSD) and 17-HSD cDNA were used to study the expression of the corresponding enzyme in the immature rat ovary during follicular development and luteinization, which were induced by treating the animals with pregnant mare serum gonadotrophin (PMSG) or with PMSG followed by human chorionic gonadotrophin (hCG). Immuno-blot analysis indicated that the Mr of the 17-HSD expressed in rat granulosa cells was 35 000, as previously shown for the human placental enzyme. In immunohistochemical studies of untreated immature rat ovaries, only the granulosa cells from small antral follicles were stained. One day after PMSG treatment, strong expression of 17-HSD was observed in the granulosa cells of growing Graafian follicles. A marked decrease in enzyme expression was observed in preovulatory follicles on day 2 of PMSG treatment, starting from the basal layers of granulosa cells and progressing toward the luminal cells. No 17-HSD expression was detected in luteinized follicles or corpora lutea 22 h after hCG injection. The stroma and theca cells were negative for 17-HSD staining. In Northern hybridization analyses, two 17-HSD mRNAs were detected (1·4 and 1·7 kb). The strongest expression for both mRNAs was detected after 1 day of PMSG treatment, coinciding with maximal immunostaining of the enzyme protein. Down-regulation of 17-HSD observed by immunohistochemistry was reflected in a similar decrease in mRNA expression and the signals were almost undetectable 22 h after hCG injection. Our data suggest that 17-HSD expression in rat granulosa cells is up-regulated during follicular development and, thereafter, the enzyme expression is down-regulated during luteinization.
Journal of Endocrinology (1994) 140, 409–417
Effect of thyroid hormones and other iodinated compounds on the transition of monocytes into veiled/dendritic cells: role of granulocyte-macrophage colony-stimulating factor, tumour-necrosis factor-α and interleukin-6
Authors: Mooij P , Simons P J , de Haan-Meulman M , de Wit H J , Drexhage H A 
Published: December 2008

Abstract
Stimulation of human peripheral blood monocytes with the thyroid hormones tri-iodothyronine (T3) and thyroxine (T4) enhanced their ability to mature into cytologically and functionally characteristic veiled/dendritic cells. Veiled/dendritic cell transition induced by T3 and T4 was dependent on the production of granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor-α (TNFα) and interleukin-6 (IL-6) in the culture, since the addition of antibodies specific for GM-CSF, TNFα and IL-6 to the culture system had blocking effects. The addition of antibodies to macrophage colony-stimulating factor and IL-1 had no effects. Contaminating T cells and B cells did not contribute to the transition of monocytes to veiled/dendritic cells, and it is therefore likely that the GM-CSF, TNFα and IL-6 produced in the culture system were derived from the monocytes themselves.
Stimulation of the blood monocytes with an optimal concentration of metrizamide (14·5%), reverse T3 (rT3; 2 × 10−10 m) or highly iodinated thyroglobulin (Tg; 2 × 10−11 m) also resulted in an increased transition of monocytes to veiled/dendritic cells, but to a lesser extent in comparison with the thyroid hormones (T3, 31 ±6% and T4, 25 ±5% vs rT3, 22 ± 8% and Tg with an iodination grade of 0·37%: 20 ± 4% veiled/dendritic cells). Administration of anti-GM-CSF, anti-TNFα and anti-IL-6 to the culture system also had blocking effects on the transition from monocytes to veiled/dendritic cells induced by the iodinated compounds. The mechanisms by which such iodinated compounds act on the monocyte to veiled/dendritic cell transition can only be speculated on (interference H2O2-generating system?).
Journal of Endocrinology (1994) 140, 503–512
The distribution of hypothalamic nitric oxide synthase mRNA in relation to gonadotrophin-releasing hormone neurons
Authors: Grossman A.B. , Rossmanith W.G. , Kabigting E.B. , Cadd G. , Clifton D. , Steiner R.A. 
Published: December 2008

ABSTRACT
Using probes for rat neural nitric oxide synthase (NOS) mRNA and GnRH mRNA, we performed in situ hybridization to survey NOS mRNA distribution within the hypothalamus of the male and female rat and sought evidence for its expression in GnRH neurons. The NOS cRNA probe was radiolabelled with 35S, and a digoxigenin-labeled rat GnRH cRNA probe was used for double-label studies. NOS mRNA was localized in discrete hypothalamic areas, in grain clusters suggestive of individual neurons. NOS mRNA-positive cells were located mainly in the supraoptic and paraventricular nucleus, particularly overlying the magnocellular division. Rostrally, cells expressing NOS mRNA were especially prominent in the diagonal band of Broca, in a distribution very similar to GnRH neurons. Nevertheless, only one of 370 cells labeled for GnRH mRNA appeared to be positive for NOS mRNA. We conclude that NOS mRNA is located prominently in regions where CRH, AVP and oxytocin cells are located. NOS mRNA-positive cells are located in close proximity to GnRH neurons, but rarely do such neurons express NOS mRNA.
ACTIVATION OF MAP KINASE ASSOCIATED WITH THE PRIMING EFFECT OF LHRH
Authors: Mitchell R. , Sim P.J. , Leslie T. , Johnson M.S. , Thomson F.J. 
Published: December 2008

ABSTRACT
A MAP kinase activity assay was developed to determine whether the LHRH receptor could activate this enzyme (particularly during LHRH priming). In anterior pituitary tissue from prooestrous rats LHRH caused concentration-dependent activation of MAP kinase after 5-10 min and continued for up to 60 min of incubation. The magnitude of this response correlated with that of LHRH priming on various days of the oestrous cycle but not with the magnitude of 1st hour (unprimed) LHRH-induced LH release. The response to LHRH was mimicked by a phorbol ester but not by ionomycin and was blocked with high potency by GF 109203X but not by H7 (in a similar manner to the PKC species that mediates LHRH priming). Neither the tyrosine kinase inhibitor lavendustin A nor the protein synthesis inhibitor cycloheximide blocked LHRH-induced MAP kinase activation. The possible functional significance of MAP kinase activation in gonadotrophs is considered with respect to LHRH priming.
Induction of ovulation by endothelin-3 in rats
Authors: Furudate S , Masaki Y , Muto T 
Published: December 2008

Abstract
Endothelins vary in their biological activity. We therefore examined the effects of endothelin-3 (ET-3) on ovulation and secretion of LH, FSH and prolactin in rats in which naturally occurring ovulation was blocked by the administration of sodium pentobarbital (40 mg/kg, i.p.) prior to the critical period (1330 h) on the day of pro-oestrus. ET-3 (10 nmol/kg) was given via the jugular vein under pentobarbital anaesthesia from 1600 to 1800 h on the day of pro-oestrous and induced ovulation in all rats whether given by venous injection or by infusion but the number of ova in rats injected with ET-3 was less than that in normally cycling control rats. Infusion of ET-3 stimulated the secretion of LH but caused a lower than expected rate of secretion of FSH. It would therefore appear that ET-3 causes release of the total amount of LH that is required for induction of ovulation. Our findings strongly suggest that ET-3 has a physiologically significant role in the regulation of anterior pituitary hormone secretion.
Journal of Endocrinology (1994) 141, 109–112
Xenopus laevis oocytes, eggs and tadpoles contain immunoactive insulin
Authors: de Pablo F , Dashner R , Shuldiner A R , Roth J 
Published: December 2008

Abstract
Insulin is a multifunctional polypeptide hormone that regulates metabolic processes and promotes mitogenesis and differentiation in vitro in the cells and tissues of several species. Its role in vivo during embryogenesis is still poorly understood. We have previously found insulin mRNA in mature Xenopus laevis oocytes and in embryos during neurulation (before organogenesis of the pancreas takes place). We have now measured insulin immunoactivity in mature oocytes, unfertilized eggs and day-2 tadpoles. Using reversed phase high performance liquid chromatography, we found low levels of insulin in extracts of oocytes (stage VI). Both Xenopus insulin I and II were detected in unfertilized eggs. The day-2 tadpoles (stages 31–33) also contained immunoactive insulin, and in swimming tadpoles (stage 46) a few clusters of cells containing insulin immunoactivity could be identified by indirect immunofluorescence. Immunoblot analysis was relatively insensitive, detecting insulin only in the adult Xenopus pancreas. In summary, insulin (from maternal origin and embryonic expression) appears to be present early enough in Xenopus laevis to influence developmental processes such as neurulation.
Journal of Endocrinology (1994) 141, 123–129
Expression of inhibin subunit mRNAs and inhibin levels in the testes of rats with stage-synchronized spermatogenesis
Authors: Klaij I A , van Pelt A M M , Timmerman M A , Blok L J , de Rooij D G , de Jong F H 
Published: December 2008

Abstract
Inhibin α- and βB-subunit mRNA expression, and levels of bioactive and immunoreactive inhibin were studied in rat testes, synchronized for the stage of the cycle of the seminiferous epithelium by treating vitamin A-deficient rats with vitamin A. Measurement of inhibin subunit mRNA expression and inhibin levels was started directly after the start of vitamin A treatment, and continued for 65 days.
Inhibin subunit mRNA expression, and testicular bioactive and immunoreactive inhibin levels increased after the start of vitamin A treatment, reaching maximum values after 9 days, when B spermatogonia and preleptotene spermatocytes had appeared in the stage-synchronized testes. The ratio between βB- and α-subunit mRNA expression was high at that time-point, whereas the ratio between bioactive and immunoreactive inhibin remained low. These data suggest a relatively high production of activin at that moment, and this may play a role in the development of B spermatogonia into preleptotene spermatocytes during the initiation of spermatogenesis.
Stage-dependency was demonstrated for inhibin subunit mRNA expression, and for the levels of bioactive and immunoreactive inhibin, in rats with complete spermatogenesis. Inhibin α-subunit mRNA expression was relatively high at stages V and XIII of the spermatogenic cycle, whereas βB-subunit mRNA expression was high at stage XIII but not at stage V. This resulted in a high βB/α subunit mRNA ratio at stage XIII. Since it has been shown that expression of the activin receptor is high at stages XIII–I, locally formed activin might play a role in the regulation of meiosis. Bioactive and immunoreactive inhibin were highly correlated during the cycle, with maximum levels at stages XIV–I.
It was concluded that the production of inhibin, and possibly activin, is dependent on the stage of the cycle of the seminiferous epithelium; these growth factors might play a paracrine role in the differentiation of spermatogenic cells.
Journal of Endocrinology (1994) 141, 131–141
Testicular and serum levels of inhibin and expression of inhibin subunit mRNAs are differentially affected by hemicastration in rats of various ages
Authors: Klaij I A , Timmerman M A , Kramer P , Meijs-Roelofs H M A , de Jong F H 
Published: December 2008

Abstract
Age-related short-term effects of hemicastration on testicular weight, serum FSH, immunoreactive inhibin, LH and testosterone, testicular levels of inhibin subunit mRNA expression, and bioactive and immunoreactive inhibin were studied in rats of 8, 15 and 22 days of age. Hemicastration led to an increased weight of the remaining testis after 24 h in 8- and 15-day-old rats, but not in 22-day-old rats. Serum FSH levels were elevated in all hemicastrated rats after 8 h. However, serum immunoreactive inhibin levels were decreased only after 72 h in 8-day-old rats and after 24 h in 15- and 22-day-old rats. Inhibin α-subunit mRNA expression was increased in the testes of hemicastrated rats of 8 and 15 days of age, whereas inhibin βB-subunit mRNA expression was elevated in the testes of 15-day-old rats but not in those of 8- and 22-day-old rats. The increase in α-subunit mRNA content per testis was caused by an increased concentration and increased testicular weight, whereas the increase in βB-subunit mRNA in the remaining testis parallelled the increased testicular weight, indicating that different mechanisms play a role in the regulation of these mRNAs. In 22-day-old rats, a transiently decreased expression of inhibin βB-subunit mRNA was observed 8 h after hemicastration. The increased inhibin α- and βB-subunit mRNA expression in 8- and 15-day-old rats did not result in increased testicular bioactive and immunoreactive inhibin content of the remaining testis, whereas in 22-dayold rats an increased immunoreactive inhibin content of the remaining testis was observed. These data indicate that efficiency of translation, post-translational modifications or transport from the testis play an important role in determining the final testicular content of inhibin.
In conclusion, the response of the remaining testis and the role of inhibin in the regulation of the pituitary-testis axis after unilateral castration depend on the age at which the animals are hemiorchidectomized.
Journal of Endocrinology (1994) 141, 143–151
The molecular basis for epitopes on the free β-subunit of human chorionic gonadotrophin (hCG), its carboxyl-terminal peptide and the hCGβ-core fragment
Authors: Dirnhofer S , Madersbacher S , Bidart J-M , Kortenaar P B W Ten , Spöttl G , Mann K , Wick G , Berger P 
Published: December 2008

Abstract
The molecular basis for antigenic determinants on the free β-subunit of human chorionic gonadotrophin (hCGβ), its carboxyl-terminal peptide (hCGβCTP) and the hCGβcore fragment (hCGβcf) was elucidated by means of monoclonal antibodies (MCAs). The objective of the present study was to resolve the antigenic topography of these three molecules in terms of epitope identification at different levels of structural organization as well as analysis of their spatial arrangement. An hCGβcf preparation, a synthetic peptide corresponding to the hCGβCTP (β109–145), overlapping synthetic peptides spanning the entire amino acid sequence of hCGβ, and a reduced and alkylated hCGβ preparation were assayed in a solid-phase one-site enzyme-linked immunoassay and in a solublephase direct-binding radioimmunoassay (RIA) or competitive RIA. The antigenic topography was mapped by incorporating the MCAs into two-site binding assays. On the surface of free hCGβ, nine different epitopes (β1–β9), arranged in three spatially distinct domains, could be distinguished. Epitopes β1–β7 were located in a single large domain on both hCGβ and the hCGβcf whereas hCGβCTP contained two topographically distant determinants, designated β8 and β9 respectively. All but the two epitopes located on hCGβCTP (β8 and β9) were destroyed by reducing and alkylating hCGβ, suggesting that most antigenic determinants are predominantly non-contiguous and require an intact tertiary structure whereas the molecular structure of hCGβCTP is linear. At a molecular level, amino acid residues spanning hCGβ 45–52, hCGβ 137–144 and hCGβ 113–116 contributed to the formation of epitopes β5, β8 and β9 respectively. We have also shown that the hCGβcf represents the immunodominant part of the free β-subunit of hCG, containing seven mainly conformationally determined epitopes, one of which has a share of the sequence β45–52. The hCGβCTP does not play a critical role in the immunologically important tertiary structure of hCGβ and was itself found to be a predominantly continuous sequence also within the native hormone, expressing two spatially distant antigenic determinants located within residues β113–116 and β137–144 respectively.
Journal of Endocrinology (1994) 141, 153–162
ACTH-induced preterm labour in the ewe is associated with increased mRNA and protein levels of myometrial gap junction protein, connexin-43
Authors: McNutt C M , Nicholson B J , Lye S J 
Published: December 2008

Abstract
The myometrial gap junction protein, connexin-43, is thought to be critical to the development of synchronous, high-amplitude contractions of the myometrium during labour. The purpose of this study was to determine the relationship between the temporal expression of myometrial connexin-43 mRNA and protein, the contractile patterns of the uterus and the changes in maternal plasma oestrogen and progesterone. On day 127 chronically catheterized fetal sheep were randomized to receive either ACTH (1 μg over 15 min every 2 h) or saline (control) infusions. Using this model, ACTH induces labour in 110 ± 5 h and produces similar endocrine profiles and changes in myometrial activity to that of term spontaneous labour. Myometrial tissue was obtained during autopsy at: 0 h (127 days: no infusion), 72 h saline, 72 h ACTH, 120 h saline, and during ACTH-induced labour (n=4/group). Northern analysis demonstrated a significant (P<0·05) increase in connexin-43 transcripts during labour (2·21±0·39; mean ±s.e.m. relative to 18S) compared with 0 h (0·67±0·17) and 72 h ACTH 0·41 ±0·11). Connexin-43 protein (as determined by Western analysis) showed a similar pattern to that of the transcripts. These changes in myometrial connexin-43 expression were associated with significant increases in the rate of rise of intrauterine pressure, frequency and maximum amplitude of uterine contractions and the maternal plasma oestrogen to progesterone ratio. No changes in connexin-43 expression, contractile parameters or endocrine profiles occurred in control animals. Our data suggest that the increased myometrial contractile coordination during preterm labour in the sheep is, at least in part, due to a steroid-associated increase in connexin-43 expression.
Journal of Endocrinology (1994) 141, 195–202