Oxytocin and/or steroid hormone binding globulin infused into the ventral tegmental area modulates progestogen-mediated lordosis.

Estradiol (E(2)) and progesterone (P(4)) have classical, steroid receptor-mediated actions in the ventral medial hypothalamus to initiate lordosis of female rodents. P(4) and the P(4) metabolite and neurosteroid, 5 alpha-pregnan-3 alpha-ol-20-one (3 alpha,5 alpha-THP), have non-classical actions in the midbrain ventral tegmental area (VTA) to modulate lordosis. We investigated the role of steroid hormone binding globulin (SHBG) and oxytocin in the VTA as mechanisms for these effects. Rats were ovariectomized and surgically implanted with bilateral guide cannulae aimed at the VTA. Rats were E(2)-primed (10 microg/0.2 ml) at hour 0, and administered 100 (Experiments 1 and 2), 500 (Experiment 3), or 0 (Experiment 1 and 4) microg/0.2 ml P(4) at hour 44. At hour 47.5, rats received bilateral infusions to the VTA, and were tested for lordosis 30 min post-infusion. Experiment 1: rats were infused with sterile saline vehicle or SHBG (4.5 pg/microl) to the VTA. SHBG, compared to vehicle, to the midbrain VTA significantly increased lordosis in E(2)- and P(4)-primed, but not E(2)-primed, rats. Experiment 2: rats were infused with bilateral infusions of sterile saline or oxytocin (1.0 pg/microl). Compared to vehicle, oxytocin to the VTA increased lordosis. Experiment 3: rats were administered bilateral intra-VTA infusions of saline or an oxytocin receptor antagonist, d(CH(2))(5),[TYr(ME)(2),Thr(4),Tyr-NH(9,2)] (1.2 pg/microl). Compared to vehicle, the oxytocin receptor antagonist to the VTA attenuated lordosis of E(2)- and P(4)-primed rats. Experiment 4: rats were E(2)-primed and infused with vehicle, oxytocin, or oxytocin antagonist. There were no effects of these manipulations in E(2)-primed rats. Thus, SHBG and/or oxytocin may have actions in the VTA for progestogen-facilitated lordosis.

Sex hormone binding globulin facilitates female sexual receptivity except when coupled to dihydrotestosterone.

Sex hormone binding globulin (SHBG) is produced in brain where it is often co-localized with oxytocin. Infusions of SHBG into the medial preoptic area-anterior hypothalamus facilitate female sexual receptivity. SHBG has receptors on plasma membranes of the prostate gland where binding of the 5alpha-reduced androgen dihydrotestosterone (DHT) by SHBG acts as an antagonist on SHBG receptors. This study attempted to determine whether pre-coupling DHT to SHBG would inhibit SHBG-induced facilitation of female sexual receptivity. Ovariectomized rats were injected daily with 0.75 microg estradiol benzoate for 3 days. On the fourth day after a pre-infusion baseline behavioral test animals were infused with 1 microl per side through bilateral cannulae with SHBG (1.77x10(-6) M), SHBG coupled to DHT (SHBG-DHT; 1.66x10(-6) M DHT), with DHT alone or with artificial cerebrospinal fluid vehicle. As before, SHBG significantly increased female sexual receptivity when infused into the medial preoptic area-anterior hypothalamus. Rats infused with SHBG-DHT had significantly lower sexual receptivity. Therefore, whereas SHBG in the medial preoptic area facilitated female sexual behavior, SHBG coupled to DHT did not. DHT itself did not significantly affect sexual receptivity. Pre-coupling DHT to SHBG eliminated the facilitative effect of SHBG on female sexual receptivity just as DHT inhibits SHBG activity at prostate SHBG receptors suggesting that central receptors for SHBG are similar to those demonstrated in the periphery.

Intravenous injection of human sex steroid hormone-binding globulin in mouse decreases blood clearance rate and testicular accumulation of orally administered [2-125I]iodobisphenol A.

Bisphenol A, an environmental compound with estrogenic activity, has been shown to bind human sex steroid hormone-binding globulin (hSHBG), the main plasma transport protein which regulates the metabolism of androgens and estrogens and limits their access to target organs. The present study was conducted to determine whether physiologically relevant concentrations of hSHBG can influence the blood clearance rate of bisphenol A and its accumulation in the testes. A radioactive [2-125I]iodobisphenol tracer was synthesized with an association constant (Ka) for binding to hSHBG of 0.14 +/- 0.01 x 10(6) M(-1) at 37 degrees C, a value much lower than for [2-125I]iodoestradiol, which was also synthesized. We used i.v. injection of immunopurified hSHBG in adult male mice to maintain hSHBG levels within the physiologically possible range for humans (27-267 nM) before gavage administration of [2-125I]iodobisphenol or [2-125I]iodoestradiol, for measuring the blood clearance rate of radioactive signal in blood samples taken during the following 120 min. Testicular accumulation of radioactivity was measured 24 h and 48 h after gavage of [2-125]iodobisphenol A. In mice receiving immunopurified hSHBG or vehicle, the time-dependent blood clearance of radioactivity exhibited a bi-exponential decrease which indicated alpha-diffusion and beta-elimination phases for both radioactive ligands. The presence of circulating hSHBG significantly and dose-dependently lowered the clearance rate of radioactivity. However, much higher circulating levels of hSHBG were required to retard the blood clearance of [2-125I]iodobisphenol A as compared to those required for [2-125I]iodoestradiol, in keeping with the important difference in their respective Ka value for binding to SHBG. In addition, mice treated with hSHBG exhibited significantly (P = 0.036) reduced testicular accumulation of radioactivity 24 h and 48 h after ingestion of [2-125I]iodobisphenol A. Provided that the binding properties of bisphenol A for hSHBG are not substantially different from those measured for [2-125I]iodobisphenol A, these findings suggest that, although hSHBG binds 2-mono-iodobisphenol A with a relatively low binding affinity, high enough concentrations of circulating hSHBG (range concentrations between 85 and 267 nM) are potentially able to exert a protective effect against exposure to bisphenol A.

Effects of sex hormone binding globulin (SHBG) on human prostatic carcinoma.

The purpose of this study was to determine what effects sex hormone binding globulin (SHBG) might have on the growth and steroid content of human prostate carcinoma. Two human prostate carcinoma cell lines were used for this study, ALVA-41 and ALVA-101. The first part of the study was to determine the effect of SHBG or albumin on the uptake of [3H]DHT in the cells. In this experiment both SHBG and albumin inhibits the uptake of [3H]DHT into each of the cell lines when studied in vitro. The degree of inhibition was dependent on the binding capacity of the protein. When [3H]thymidine uptake was measured in each of the cell lines following either the addition of SHBG or albumin to the culture media, an increase in uptake and presumably DNA synthesis was noted in the ALVA-41 and ALVA-101 cells for SHBG additions but not for albumin. Further, this stimulation was increased when testosterone was added to the media, however, [3H]thymidine uptake was decreased by high concentrations of dihydrotestosterone (DHT) or if the SHBG was saturated with DHT prior to being added to the media. The cells also demonstrate high affinity cell membrane receptors for SHBG. Finally, using a 3', 550 bp cDNA or SHBG, 1.9 and 2.8 kb mRNAs were detected on Northern analysis of the ALVA-101 and ALVA-41 cells. These data indicate SHBG can inhibit uptake of steroids into the prostate, but also it may act as a stimulus for growth through a SHBG cell surface receptor. In addition, the growth effect may be through an autocrine effect from SHBG or a SHBG-related peptide.

Reduction of testosterone availability to 5 alpha-reductase by human sex hormone-binding globulin in the rat ventral prostate gland in vivo.

The present studies assess the effects of human sex hormone-binding globulin (SHBG) on the conversion of [3H]testosterone (T) into dihydrotestosterone (DHT) in rat ventral prostate gland in vivo using a constant aortic infusion technique. The DHT/T ratio was determined using two-dimensional thin-layer chromatography (TLC), and these results were confirmed with reverse-phase high-performance liquid chromatography. The prostatic gland DHT/T ratio was 2.1 +/- 0.4, 1.3 +/- 0.2, 0.24 +/- 0.02, or 1.1 +/- 0.2, following a 60 sec aortic perfusion of [3H]testosterone dissolved in either Krebs-Henselite buffer (KHB), 5 g/dl human serum albumin (HSA), human pregnancy serum (HPS), or heat inactivated HPS, respectively. Heat inactivation (60 degrees C, 60 min) selectively denatured SHBG in HPS. The distribution of [3H]testosterone in rat ventral prostate was examined with thaw-mount light in microscopic autoradiography. Following an aortic perfusion of [3H]testosterone in buffer alone, the radiolabeled steroid was uniformly distributed among the epithelial and stromal compartments. However, the [3H]steroid hormone was selectively sequestered in the stromal compartment following aortic perfusion of HPS. In conclusion, these studies demonstrate that human SHBG markedly restricts the availability of circulating testosterone to 5 alpha-reductase in the prostate gland in vivo and that the presence of SHBG in serum causes the selective sequestration of the steroid hormone within the stromal compartment of the gland in vivo.