Margaret Yu

Heterogeneity of [3H]estradiol binding sites in the rat prostate: properties and distribution of type I and type II sites.

In order to assess the rat prostate as a target tissue for receptor-mediated estrogen action, we have studied the properties and distributions of estrogen binding sites in the dorsolateral (DLP) and ventral (VP) prostate. Saturation analyses over a wide range of [3H]estradiol ([3H]E2) concentrations (0.5-100 nM) revealed two distinct types of binding sites in the cytosol and nuclear fractions of DLP of intact rats. The high affinity (type I) estrogen binding sites saturated at 2-4 nM of [3H]E2 and had a capacity of 170 fmol/mg DNA in the cytosol and 400 fmol/mg DNA in the nuclei. DLP type I sites had ligand specificity similar to that described for the classical estrogen receptors (ERs) found in female target tissues. The moderate affinity (type II) estrogen binding sites saturated at 15-30 nM of [3H]E2 and had a capacity of 850 fmol/mg DNA in the cytosol and 1600 fmol/mg DNA in the nuclei. DLP type II sites shared some characteristics of the type II ERs described for the rat uterus; they were estrogen specific, heat labile, and sensitive to reducing agents such as dithiothreitol. Saturation analyses on VP cytosols and nuclear fractions revealed only high affinity sites but no moderate affinity sites in the tissue preparations. Our finding that prostatic type II estrogen binding sites are present exclusively in the DLP supports the concept that basic biological differences exist between the two major prostatic lobes of the rat. Furthermore, our findings may help elucidate the observed differences in susceptibility between these two lobes to the hormonal induction of proliferative prostatic lesions.

Decreased Production of Transforming Virus and Altered Antigenic Behaviour in Cultured Avian Sarcoma Cells

Tumours induced in chickens by inoculation of avian sarcoma viruses are frequently capable of undergoing spontaneous regression. It is only those tumour cells which have been derived from progressively growing neoplasms that are able to produce transforming progeny virus in vitro and to shed into the culture medium antigens which are specifically reactive with the peripheral lymphocytes of sarcoma-bearing hosts. Following multiple passages and extended growth in culture, however, the ability of these tumour cell fluids to stimulate the lymphocytes of sensitized hosts diminishes in concert with the declining capacity of these cells to continue to synthesize fully transforming progeny virus. In certain instances, however, aged tumour cells are able to synthesize particles which contain the enzyme RNA-dependent DNA polymerase yet lack detectable envelope glycoprotein.

The Conjoint Actions of Androgens and Estrogens in the Induction of Proliferative Lesions in the Rat Prostate

Despite extensive past investigations, the etiology of human prostatic carcinoma remains largely undefined. It has been suggested that age-associated changes in the sex hormone milieu may be the common cause of prostatic cancer development in man Several lines of evidence furnish support for this credence. First, circulating levels of free testosterone (T) in men decline with age while those of free 17s-estradiol (E2) remain unchanged, thus resulting in a much higher ratio of free E2 to T in older men (1,2). Second, it has been reported that among black Americans who have the highest incidence of prostatic cancer in the world, both plasma free T and estrone levels are elevated at young age (3). Taken together, these findings suggest that changing levels of circulating androgens and/or estrogens or an altered ratio of the two classes of steroids in plasma are important determinants of prostatic carcinogenesis in the human male. Moreover, the influence of sex hormones could begin in the early decades of a man’s life and have a long-lasting impact on the prostate.

Hormonal regulation of nuclear type II estrogen binding sites in the dorsolateral prostate of noble rats

We previously demonstrated that simultaneous treatment of Noble (NBL) rats with estradiol (E2) and testosterone (T) for 16 weeks induces a proliferative response selectively in the dorsolateral prostates (DLP) of all treated animals [1, 2]. The unique sensitivity of rat DLP to the conjoint androgen-estrogen-induced mitogenic action may be attributable to the presence of a moderate affinity, high capacity, nuclear estrogen binder (type II sites) found exclusively in this prostatic lobe [2, 3]. Little is known about whether prostatic type II site levels are under hormonal regulation. The aim of this study is to determine whether testicular steroids play a role in regulating the basal and/or induced levels of type II site expression in rat DLP. In the first experiment, rats were castrated and immediately treated with 5 alpha-dihydrotestosterone (DHT) and/or E2 for 6 weeks to determine whether these steroids, separately or jointly, could sustain DLP type II site levels in castrates. Treatments of castrated rats with DHT and DHT+E2 were found to be effective in maintaining DLP type II site levels and gland wet weights at values close to those found in intact untreated controls, while treatments with E2 failed to maintain these parameters at levels observed in intact animals. In the second experiment, intact rats were treated with an androgen (T or DHT) or E2, alone or in combination, for 16 weeks to ascertain which hormonal regimen could induce a higher level of type II site expression in the DLP. Treatments of rats with an androgen (T or DHT) or E2 alone did not change DLP type II site levels even though T treatment caused a slight increase in gland weight, while E2 treatment induced prostatic atrophy. Contrary to single hormone treatments, combined T + E2 and DHT+E2 treatments were effective in inducing a doubling of type II sites and increases in wet weight of the DLPs. These data indicate that testicular androgen is the primary factor responsible for maintaining a basal level of type II site expression in rat DLP, while conjoint androgenic-estrogenic action is needed for the induction of a higher level of type II site expression in the tissue.

Seasonal variation in hepatic binding of estrogen in the turtle, Chrysemys picta.

The present study describes the seasonal changes of the estrogen receptor (ER) system in the liver of the turtle, Chrysemys picta. [3H]Estradiol ([3H]E2) binding capacities and affinities of liver cytosols and nuclear extracts were measured with established procedures and analyzed by Scatchard plots. Our data revealed significant seasonal variations in both receptor content and the ERs affinity for [3H]E2 in the liver of the turtle. Nuclear ER content remained at a fairly stable level of 70 fmol/g tissue throughout the year, but exhibited two sharp increases in the months of May and October, attaining values of approximately 150 fmol/g tissue. These rises in nuclear ER content coincided temporally with the two peaks of vitellogenin accumulation and estrogen surge in the annual cycle previously reported for the female turtle. Cytosolic ER level in the liver exhibited a much more complex pattern. It fluctuated with a much wider range of 80 to 250 fmol/g tissue. Right after oviposition in July, it declined to its lowest value of 80 +/- 9 fmol/g tissue but gradually increased to 160 +/- 15 fmol/g tissue in September and remained at this medium range for the remaining of the fall season but rose sharply again to the highest value of the year in December, reaching a value of 270 +/- 7 fmol/g tissue. In the following winter months, cytosolic ER level declined until March before it rebounded to a second peak value of 242 +/- 6 fmol/g tissue in May.(ABSTRACT TRUNCATED AT 250 WORDS)

High-affinity binding of [3H]estradiol-17β by an estrogen receptor in the liver of the turtle

Specific [3H]estradiol-17 beta ([3H]E2) binding activity (EBA) with characteristics of an estrogen receptor (ER) was demonstrated in cytosols and nuclear extracts of the female turtle, Chrysemys picta. Three different receptor assays (dextran-coated charcoal assay, hydroxylapatite batch procedure, and DNA-cellulose chromatography) were evaluated in terms of their applicability in analyzing large numbers of samples. For the measurement of cytosolic EBA, the hydroxylapatite batch procedure was found to be the most reliable assay. On the other hand, the dextran-coated charcoal assay was found to be the most appropriate method for the measurement of nuclear EBA. Turtle hepatic EBA binds [3H]E2 with high affinity (cytosolic, 17.4 +/- 2.8 X 10(9) M-1; nuclear, 17.7 +/- 1.9 X 10(9) M-1), limited capacity (cytosolic, 133.7 +/- 4.6 fmol/g tissue; nuclear, 81.1 +/- 9.0 fmol/g tissue), and strict steroid specificity. The EBA bound natural estrogens (E2, estrone, estriol) as well as the nonsteroidal estrogen, diethylstilbestrol, but exhibited little affinity for androgens, progesterone, or corticosterone. The turtle hepatic EBA resembled mammalian and avian ERs in terms of binding characteristics; however, unlike mammalian and avian ERs it was shown to be heat-labile. Incubation at 30 degrees caused rapid loss of [3H]E2 binding activity in both cytosolic and nuclear fractions. The exchange between [3H]E2 and the endogenously bound estrogen was slow at 4 and 15 degrees, but the exchange process was facilitated in the presence of the chaotropic salt, NaSCN. Establishment of quantitation methods for both cytosolic and nuclear forms of EBA will enable future investigation of the mechanism and regulation of estrogen action in the liver of this turtle species.

Effects of hypophysectomy and ovariectomy on hepatic estrogen receptor content in the turtle, Chrysemys picta☆

Estrogen receptor (ER) content in the cytosol and nuclear extract of the liver of adult female Chrysemys picta (control females) was measured in February, May, early June, and late June. Both cytosolic (C) and nuclear (N) hepatic ER content rose from low levels (C: 189 +/- 19; N: 52 +/- 23 fmol/g tissue) in February to significantly higher levels in May (C: 437 +/- 15; N: 124 +/- 25 fmol/g tissue) before declining to lower values in early June (C: 298 +/- 19; N: 118 +/- 20 fmol/g tissue) and late June (C: 274 +/- 22; N: 89 +/- 13 fmol/g tissue). This pattern of seasonal variation in hepatic ER content was also observed in gonadectomized females. Levels of hepatic ER content in ovariectomized turtles fluctuated in the same manner as those found in control females. However, elevations in hepatic ER contents were not observed in hypophysectomized females. Removal of the pituitary resulted in a drastic reduction in the May value (316 +/- 49 fmol/g tissue) of cytosolic ER content and a complete blockade in a rise of nuclear ER content in the liver of the turtle. Nuclear hepatic ER content remained at around 50 fmol/g tissue throughout the months of May and June. The two surgeries procedures caused no significant changes in the ERs affinity for estrogen. The present findings demonstrate a direct control of the pituitary over hepatic ER content which signifies important pituitary regulation of the vitellogenic process, at the liver level, during the spring reproductive season.