Gonadal steroid modulation of neuroendocrine transduction: A transynaptic view

Summary 1. Steroid hormones act on neuronal communication through different mechanisms, ranging from transynaptic modulation of neurotransmitter synthesis and release to development and remodeling of synaptic circuitry. Due the wide distribution of putative brain targets for steroid hormones, acute or sustained elevations of their circulating levels may affect, simultaneously, a variety of neuronal elements. In an elementary mode of interaction, steroids are able to modulate both the synthesis and release of a neurotransmitter at a particular synapse, and the response of its target postsynaptic cells. Using two neuroendocrine transducing systems—the rat pineal gland and the GT_1–7 cell line—we have examined these interactions and the following findings are discussed in this article.2. In the rat, pineal melatonin production is partially controlled by gonadal hormones. In females, melatonin synthesis and secretion is reduced during the night of proestrus, apparently as a consequence of elevated estradiol and progesterone levels. In males, circulating testosterone seems to be necessary to maintain the amplitude of the nocturnal melatonin peak.3. Some gonadal effects on pineal activity are exerted on its noradrenergic input, since changes in circulating steroid hormone levels are able to induce acute modifications of tyrosine hydroxylase activity in pineal sympathetic nerve terminals.4. Gonadal steroids are also able to regulate the response of pineal cells to adrenergic stimulation, since in vivo treatment of both male and female rats with steroid hormone blockers induces profound modifications in adrenergically-induced accumulation of cyclic AMP (cAMP) in dispersed pinealocytes.5. Direct exposure of pineal cells from gonadectomized female and male rats to estradiol (E₂) or testosterone (T), respectively, potentiates pinealocyte response to adrenergic activation. In addition, short-term (15 min) exposure to either progesterone (Pg) or progesterone coupled to bovine serum albumin (P-3-BSA) suppresses the E₂-dependent potentiation of adrenergic response in female rat pinealocytes.6. Exposure of GT_1–7 cells to E₂ completely blocked the norepinephrine (NE)-induced elevation of cAMP content. In E₂-treated GT_1–7 cells, additional exposure (15 min) to either Pg or P-3-BSA abolished E₂-dependent inhibition of NE responsiveness. In addition, P-3-BSA alone increased basal cAMP levels in GT_1–7 cells, regardless previous exposure to E₂.7. In conclusion, there are evidences, both from the current literature and from the present results, supporting the view that in some neuroendocrine systems gonadal hormones modulate neurotransmission by acting, simultaneously, at pre- and postsynaptic sites. The models presented here constitute appropriate examples of this transynaptic mode of steroid action and, therefore, may offer a useful approach to investigate steroid hormone actions on the brain.     

A highly potent series of fluoroalkyl benzoxazine pyridine-N-oxide potassium channel openers

A new structural class of fluoroalkyl benzoxazine pyridine-N-oxide potassium channel openers (PCOs) is described.     

Luteal inadequacy during the early luteal phase of subfertile cows

A study was made of early luteal function (up to Day 6) in cyclic and pregnant heifers and also in older, subfertile cows. There were no differences in vivo or in vitro between cyclic and pregnant heifers, indicating no luteotrophic effect of the embryo at this stage, but the increase in postovulatory peripheral progesterone concentrations was delayed (P less than 0.01) and occurred more slowly (P less than 0.001) in the subfertile cows than in the heifers. The corpora lutea of the subfertile cows were heavier (P less than 0.001) than those of the heifers on Day 6. Basal progesterone production by dispersed luteal cells was similar between heifers and subfertile cows, but there was a difference (P less than 0.001) in the pattern of response to exogenous LH and PGE-2. Cells from subfertile cows were less sensitive to the stimulatory effects of PGE-2 and although LH increased (P less than 0.001) progesterone production by all cells, this stimulation by a low dose of LH was inhibited by PGE-2 in luteal cells from subfertile cows. This effect did not occur in the luteal cells from heifers. These results indicate the possibility that luteal inadequacy, due to a diminished response to circulating luteotrophic hormones, may contribute to embryo mortality in subfertile cows.     

Day-Night Rhythm of Rat Pineal Tyrosine Hydroxylase Activity as Determined by HPLC with Amperometric Detection

Tyrosine hydroxylase activity in the rat pineal gland was measured by means of HPLC determination of the amount of L-3,4-dihydroxyphenylalanine formed. Enzyme activity showed a clear day-night rhythm, paralleling that of plasma melatonin levels in the same animals, with values being high during the dark period apparently because of changes in Vmax. In animals maintained under constant illumination for 3 days, tyrosine hydroxylase activity and plasma melatonin level rhythms were completely abolished, a result indicating that both are under photoperiodic control.     

Manipulation of aggressive behavior in adult DBA/2/Bg and C57BL/10/Bg male mice implanted with testosterone in silastic tubing

Adult DBA/2/Bg and C57BL/10/Bg male mice were orchidectomized and implanted subcutaneously with one of two doses of testosterone (T) in Silastic tubing. Steroid doses which resulted in significant atrophy and hypertrophy of accessory sex tissues were determined for each strain. Relative to sham-castrated vehicle-implanted controls, both hypophysiological and hyperphysiological implants caused augmentation of aggression in C57BL/10/Bg males. In DBA/2/Bg mice, the hypophysiological dose reduced and the hyperphysiological implant increased aggression scores. The results demonstrate that agonistic behavior in male mice is influenced by the titer of circulating T and emphasize the importance of genetic variance in the study of hormones and behavior.     

Measuring the structural impact of mutations on cytochrome P450 21A2, the major steroid 21-hydroxylase related to congenital adrenal hyperplasia

Abstract

Congenital adrenal hyperplasia is an inherited autosomal recessive disorder related to deficient cortisol synthesis. The deficiency of steroid 21-hydroxylase (cytochrome P450 21A2), an enzyme involved in cortisol synthesis, is responsible for ～95% of cases of congenital adrenal hyperplasia. This metabolic disease exhibits three clinical forms: salt-wasting, simple virilizing, and non-classical form, which are divided according to the degree of severity. In the present study, structural and mutational analyses were performed in order to identify the structural impact of mutations on cytochrome P450 21A2 and correlate them with patient clinical severity. The following mutations were selected: arginine-356 to tryptophan (R356W), proline-30 to leucine (P30L), isoleucine-172 to asparagine (I172N), valine-281 to leucine (V281L), and the null mutation glutamine-318 (Q318X). Our computational approach mapped the location of residues on P450 and identified their implications on enzyme electrostatic potential mapping to progesterone and heme binding pockets. Using molecular dynamics simulations, we analyzed the structural stability of ligand binding and protein structure, as well as possible conformational changes at the catalytic pocket that leads to impairment of enzymatic activity. Our study sheds light on the impact structural mutations have over steroid 21-hydroxylase structure-function in the cell.

Communicated by Ramaswamy H. Sarma     

Antimicrobial, antitumor and antileishmania screening of medicinal plants from Guinea-Bissau.

Following an ethnobotanical search carried out in Guinea-Bissau, eighteen extracts derived from sixteen medicinal species were screened for antimicrobial, antitumor and antileishmania activity. Significant antitumor activity was found for Holarrhena floribunda against KB (squamous carcinoma), SK-Mel 28 (melanoma), A 549 (lung carcinoma) and MDA-MB 231 (mamma carcinoma) cell lines, with corresponding IC50 values of 7.9, 9.0, 3.4 and 9.9 micrograms/ml. Khaya senegalensis and Anthostema senegalense exhibited a significant activity against Leishmania donovani with IC50 values of 9.8 and 9.1 micrograms/ml, respectively. Most of the extracts showed week or moderate antibacterial and antifungal activity, with MIC values in the range 0.25-1.0 mg/ml. Active extracts were submitted to bioassay-guided fractionation, and the IC50 and MIC of the active fractions were determined.