雄激素受体对真核基因转录的调控

雄激素受体是典型的核受体,它对真核基因转录的调控作用受到日益广泛的重视。本文主要阐述了雄激素受体的分子结构,重点总结了雄激素受体介导真核基因转录起始的过程,概述了激素受体辅助使用因子及受体的核转运等受体功能的调控,这些是进一步研究真核基因表达调控机制及治疗雄激素相关疾病的理论基础。     

Enhancement of Declarative Memory: From Genetic Regulation to Non-invasive Stimulation

The problem of memory enhancement is extremely important in intellectual activity areas and therapy of different types of dementia, including Alzheimer’s disease (AD). The attempts to solve this problem have come from different research fields. In the first part of our review, we describe the results of targeting certain genes involved in memory-associated molecular pathways. The second part of the review is focused on the deep stimulation of brain structures that can slow down memory loss in AD. The third part describes the results of the use of non-invasive brain stimulation techniques for memory modulation, consolidation, and retrieval in healthy people and animal models. Integration of data from different research fields is essential for the development of efficient strategies for memory enhancement.     

Androgen receptor phosphorylation. Regulation and identification of the phosphorylation sites

Activation of signal transduction kinase cascades has been shown to alter androgen receptor (AR) activity. Although it has been suggested that changes in AR phosphorylation might be directly responsible, the basal and regulated phosphorylations of the AR have not been fully determined. We have identified the major sites of AR phosphorylation on ARs expressed in COS-1 cells using a combination of peptide mapping, Edman degradation, and mass spectrometry. We describe the identification of seven AR phosphorylation sites, show that the phosphopeptides seen with exogenously expressed ARs are highly similar to those seen with endogenous ARs in LNCaP cells and show that specific agonists differentially regulate the phosphorylation state of endogenous ARs in LNCaP prostate cancer cells. Treatment of LNCaP cells with the synthetic androgen, R1881, elevates phosphorylation of serines 16, 81, 256, 308, 424, and 650. Ser-94 appears constitutively phosphorylated. Forskolin, epidermal growth factor, and phorbol 12-myristate 13-acetate increase the phosphorylation of Ser-650. The kinetics of phosphorylation of most sites in response to hormone or forskolin is temporally delayed, reaching a maximum at 2 h post-stimulation. The exception is Ser-81, which continues to display increasing phosphorylation at 6 h. These data provide a basis for analyzing mechanisms of cross-talk between growth factor signaling and androgen in prostate development, physiology, and cancer.     

Development of NADPH-Diaphorase in the Avian Retina: Regulation by Calcium Ions and Relation to Nitric Oxide Synthase

Abstract: Nitric oxide plays an important role as an intercellular messenger in the CNS. In the present work we measured NADPH-diaphorase activity, which is considered to be a marker of cells producing nitric oxide, in homogenates of the developing chick retina. The enzyme activity can be detected beginning in 8-day-old embryonic retinas with no further quantitative variations throughout development. Arginine analogues inhibit ∼65% of the activity in embryonic retinas and 50% in posthatched retinas. The enzyme is stimulated 50% by 2 m M calcium chloride in retinas from 8 to 14 embryonic days, but this effect decreases to 20% in 17-day embryonic retinas and practically disappears in posthatched animals. The stimulation by calcium is completely blocked by arginine analogues. The decrease in enzyme activity at posthatched retinas is not due to stimulation by endogenous calcium or the presence of insufficient amounts of calmodulin, because addition of EGTA or calmodulin, respectively, did not restore the stimulation to levels observed at embryonic stages. Inhibition of NADPH-diaphorase activity by N ^G-nitro- l -arginine or l - N ^G-(iminoethyl)ornithine is concentration dependent with IC₅₀ values of ∼1 m M at all stages studied. However, in the presence of calcium, the inhibition by both analogues is shifted to the left and is apparently biphasic at all developmental stages, including in posthatched animals, with IC₅₀ values in the low micromolar range. NADPH-diaphorase was also detected by histochemistry in specific groups of cells in the early embryonic retina and in subsets of amacrine and ganglion cells, as well as in photoreceptors, in more developed retinas. The results indicate that different isoforms of nitric oxide synthase are present in the chick retina and that a calcium-dependent isoform is predominant in early periods of development.     

Gene Targeting Approaches to Neuroendocrinology: Oxytocin, Maternal Behavior, and Affiliation

Transgenic technology affords exciting new opportunities in the field of behavioral neuroendocrinology. We have extended our research into the behavioral function of oxytocin in maternal and social behavior using two transgenic approaches: (i) targeted deletion of the oxytocin gene in mice and (ii) augmented oxytocin receptor expression in the brain. Mice genetically deficient in oxytocin can mate, give birth, and display normal maternal behavior; however, milk ejection and certain aspects of social behavior are affected. Comparative studies of oxytocin receptors have led to the observation that species differences in social organization are associated with differences in receptor distribution. Specifically, monogamous prairie voles and nonmonogamous, asocial montane voles exhibit different patterns of OT receptor expression in the brain. Transgenic mice have been created with a reporter gene driven by the prairie vole oxytocin receptor gene promoter. Analysis of the expression pattern suggests that it should be possible to manipulate receptor expression in the vole brain in order to examine the effects of receptor distribution on behavior.     

Neuroendocrinology and its Quantitative Development: A Bioengineering View

Honor thy father and thy mother, say the Holy Scriptures [1], for they at least gave thee this biological life, but honor thy teachers, too, for they gave thee knowledge and example.     

Oxygen and CO2 Exchange and Acid-Base Regulation in the Avian Embryo

The avian embryo exchanges the oxygen and carbon dioxide withthe ambient air by diffusion. The respiratory organ is the chorioallantois,endowed with a rich circulation. Between ambient air and chorioallantoiccapillary blood are interposed the porous shell fibrous shellmembranes, and the chorioendothelium which compose the diffusionbarrier. The air cell is formed between the two shell membranesin the blunt end of the egg. The diffusion barrier is dividedinto an outer barrier (shell plus outer membrane) and an innerbarrier (inner membrane plus chorioendothelium and capillaryblood). The resistance to gas diffusion (the reciprocal of thediffusive conductance) in the outer barrier is almost fixedthroughout incubation while that in the inner barrier decreasesas the embryo develops. Because of the fixed outer barrier conductance,the embryo is obliged to take up oxygen under hypoxic conditionsagainst increasing metabolism with development and encountersa relative respiratory acidosis. In connection with the diffusivehypoventilation caused by the fixed outer barrier conductancethe respiratory factors of the allantoic circulation changeprogressively with development to moderate the restraint ofgas exchange through the shell. Blood oxygen capacity and hemoglobinincrease with development in association with an increase inerythrocyte count and hematocrit value. In addition, a progressiveleftward shift of the oxygen dissociation curve occurs. Theincreases in the allantoic blood flow and chorioallantoic capillaryvolume contribute to the increasing conductance of the innerbarrier. Furthermore regulation of acid base balance is inferredin the developing embryo.     

Regulation of aldosterone production by ion channels: From basal secretion to primary aldosteronism

Aldosterone is produced by zona glomerulosa (ZG) cells of the adrenal cortex and plays a key role in balancing water and electrolytes levels. Autonomous overproduction of aldosterone leads to primary aldosteronism (PA), which is the most common form of secondary endocrine hypertension. Recently, significant progress has been made towards understanding the genetic basis of PA, where increasing clinical evidence suggests that mutations in ion channels appear to be the major cause of aldosterone-producing adenomas. In this review, we focused on potassium and calcium channels that regulate aldosterone secretion, and their roles in the pathology of PA. Because potassium and calcium channels are differentially expressed in ZG cells in different species of mammals, the limitations of published studies are also discussed.     

Developmental Regulation of Phenylethanolamine N-Methyl Transferase in Avian Sympathetic Nerves

The presence of the catecholamine synthetic enzyme, phenylethanolamine N-methyl transferase (PNMT), has been detected in the expansor secundariorum, a smooth muscle of the avian wing. The concentration of the enzyme was estimated over a 10-week time course from 17 days incubation to 9 weeks posthatch and found to increase rapidly up until hatch in parallel with dopamine beta-hydroxylase activity, but then to fall precipitously to very low levels. The time course of the initial increase in activity corresponds to the presence of ingrowing sympathetic nerve fibres, and denervation of the expansor results in loss of greater than 80% of the PNMT activity. It is concluded that during the period of innervation the growing nerves contain the enzyme PNMT and therefore have the capacity to synthesize adrenaline, but that shortly after innervation is complete the capacity to synthesize adrenaline is lost. Several alternate mechanisms are proposed to explain the observations.     

Estrogen Regulation of Yolk and Non-Yolk Protein Synthesis in the Avian Liver

The effects of acute and chronic estrogen treatment on two egg yolk proteins, vitellogenin and apoVLDL-II, and two non-yolk proteins, ovalbumin and apoA-I, were studied by immunocytochemical techniques. Three groups of cockerels received either no treatment, or a single injection of diethylstilbestrol (DES, 2.5 mg) (acute stimulation) 24 h before killing, or 14 daily injections of 2.5 mg DES (chronic stimulation) before killing. The animals were killed at 4 weeks of age and their livers examined with respect to the distribution of the four different proteins by the indirect immunoperoxidase method. Vitellogenin was undetectable in the untreated cockerel liver. A single injection of DES resulted in the appearance of the protein in approximately 10%–15% of the hepatocytes. Chronic DES stimulation increased the number of positive cells to about 20%. In contrast, apoVLDL-II was present in 1%–2% of the hepatocytes in untreated animals. It was detected in an increased proportion (20%–25%) of cells after a single dose of DES. After chronic estrogen treatment, there was a very marked increase in the number of positive cells (> 90%). Ovalbumin was undetectable in untreated cockerel liver, while apoA-I was detected in an extremely low proportion of cells (0.005%–0.01%). Neither ovalbumin nor apoA-I distribution seemed to be affected by a single dose of DES. However, chronic DES treatment resulted in the appearance of ovalbumin-containing cells (∼ 0.02%) and a marked increase in the number of cells containing apoA-I (10%–15%).     

Regulation of Male Sexual Behavior by Progesterone Receptor, Sexual Experience, and Androgen

Recent studies have demonstrated that physiological doses of progesterone may facilitate the androgen-dependent display of male sexual behavior in laboratory rats and three species of lizard. We used mice with a targeted disruption of the progesterone receptor to investigate whether such interactions exist in male mice and whether they may be modified by sexual experience. We found that naive intact male progesterone receptor knockout (PRKO) mice exhibit reduced mount frequencies compared to wild-type (WT) mice. Also unlike WT mice, sexually experienced PRKO males show profound losses in many measures of sexual behavior following castration. In a second experiment, we tested whether male mice heterozygous for a null mutation at the progesterone receptor locus were responsive to testosterone and progesterone treatment. We found that heterozygous males showed a reduced response to testosterone. The data are consistent with experiments indicating that the progesterone receptor is able to facilitate male-typical sex behaviors in other species and suggest novel mechanisms underlying the interaction of androgens and experience.     

Regulation of Endochondral Cartilage Growth in the Developing Avian Limb: Cooperative Involvement of Perichondrium and Periosteum

The perichondrium and periosteum have recently been suggested to be involved in the regulation of limb growth, serving as potential sources of signaling molecules that are involved in chondrocyte proliferation, maturation, and hypertrophy. Previously, we observed that removal of the perichondrium and periosteum from tibiotarsi in organ culture resulted in an overall increase in longitudinal cartilage growth, suggesting negative regulation originating from these tissues. To determine if the perichondrium and periosteum regulate growth through the production of diffusible factors, we have tested various conditioned media from these tissues for the ability to modify cartilage growth in tibiotarsal organ cultures from which these tissues have been removed. Both negative and positive regulatory activities were detected. Negative regulation was observed with conditioned medium from (1) cell cultures of the region bordering both the perichondrium and the periosteum, (2) co-cultures of perichondrial and periosteal cells, and (3) a mixture of conditioned media from perichondrial cell cultures and periosteal cell cultures. The requirement for regulatory factors from both the perichondrium and periosteum suggests a novel mechanism of regulation. Positive regulation was observed with conditioned media from several cell types, with the most potent activity being from articular perichondrial cells and hypertrophic chondrocytes.     

Conversion of Androgen Receptor Signaling From a Growth Suppressor in Normal Prostate Epithelial Cells to an Oncogene in Prostate Cancer Cells Involves a Gain of Function in c-Myc Regulation

In normal prostate, androgen-dependent androgen receptor (AR) signaling within prostate stromal cells induces their secretion of paracrine factors, termed “andromedins” which stimulate growth of the epithelial cells. The present studies demonstrate that androgen-dependent andromedin-driven growth stimulation is counter-balanced by androgen-induced AR signaling within normal adult prostate epithelial cells resulting in terminal G₀ growth arrest coupled with terminal differentiation into ΔNp63-negative, PSA-expressing secretory luminal cells. This cell autonomous AR-driven terminal differentiation requires DNA-binding of the AR protein, is associated with decreases in c-Myc m-RNA and protein, are coupled with increases in p21, p27, and SKP-2 protein expression, and does not require functional p53. These changes result in down-regulation of Cyclin D₁ protein and RB phosphoryation. shRNA knockdown documents that neither RB, p21, p27 alone or in combination are required for such AR-induced G₀ growth arrest. Transgenic expression of a constitutive vector to prevent c-Myc down-regulation overrides AR-mediated growth arrest in normal prostate epithelial cells, which documents that AR-induced c-Myc down-regulation is critical in terminal growth arrest of normal prostate epithelial cells. In contrast, in prostate cancer cells, androgen-induced AR signaling paradoxically up-regulates c-Myc expression and stimulates growth as documented by inhibition of both of these responses following exposure to the AR antagonist, bicalutamide. These data document that AR signaling is converted from a growth suppressor in normal prostate epithelial cells to an oncogene in prostate cancer cells during prostatic carcinogenesis and that this conversion involves a gain of function for regulation of c-Myc expression.     

Regulation and Methylation of Tumor Suppressor MiR-124 by Androgen Receptor in Prostate Cancer Cells

Prostate cancer (PCa) is the most frequently diagnosed cancer for men in the developed world. Androgen receptor signaling pathway plays an important role in prostate cancer progression. Recent studies show that microRNA miR-124 exerts a tumor suppressive function in prostate cancer. However, the relationship between AR and miR-124 is unclear. In the present study, we found a negative feedback loop between AR and miR-124 expression. On one hand, miR-124 was a positively regulated target gene of the AR, on the other hand, overexpression of miR-124 inhibited the expression of AR. In addition, we found that miR-124-2 and miR-124-3 promoters were hypermethylated in AR-negative PCa cells. Furthermore, overexpression of miR-124 inhibited proliferation rates and invasiveness capacity of PCa cells in vitro, and suppressed xenograft tumor growth in vivo. Taken together, our results support a negative feedback loop between AR and miR-124 expression. Methylation of miR-124-2 and miR-124-3 may serve as a biomarker for AR-negative PCa cells, and overexpression of miR-124 might be of potential therapeutic value for the treatment of PCa.