William R. Crowley

Neuropeptide Y and peptide YY neuronal and endocrine systems

An extensive system of neuropeptide Y (NPY) containing neurons has recently been identified in the central and peripheral nervous system. In addition, NPY and a structurally related peptide, peptide YY (PYY), containing endocrine cells have been identified in the periphery. The NPY system is of particular interest as the peptide coexists with catecholamines in the central and sympathetic nervous system and adrenal medulla. Evidence has been presented which indicates that NPY may play important roles in regulating autonomic function.

Evidence that NPY-containing neurons in the brainstem project into selected hypothalamic nuclei: implication in feeding behavior

Recent studies show that bilateral neural transections (NT) at the level of dorsal tegmentum in the mesencephalon significantly increase food intake and decrease latency to onset of feeding behavior in response to neuropeptide Y (NPY). The increased responsiveness to NPY may be due to denervation-induced hypersensitivity to NPY in hypothalamic sites that mediate feeding behavior in rats. To test this hypothesis we have studied the effect of NT on NPY concentrations in 7 neural sites of male rats. Two weeks after NT, NPY levels in 3 hypothalamic nuclei--suprachiasmatic nucleus, arcuate nucleus and ventromedial hypothalamus--were not altered by NT thereby suggesting that NPY innervations in these nuclei may be derived mainly from NPY perikarya in the ARC and elsewhere in the diencephalon. On the other hand, NPY concentrations were markedly decreased (50-60%) in the medial preoptic area, paraventricular nucleus, median eminence and dorsomedial nucleus indicating that a substantial number of neurons in the brainstem, which show coexistence of NPY and adrenergic transmitters, project into these 4 diencephalic nuclei. These findings indicate that NPY-containing neurons in the brainstem may project into selected hypothalamic sites and the reduction in the NT rats of NPY levels, especially in the paraventricular nucleus, may be responsible for the reported increase in sensitivity of the NPY-induced feeding response.

Neuropeptide Y Stimulates the Release of Luteinizing Hormone-Releasing Hormone from Medial Basal Hypothalamus in vitro: Modulation by Ovarian Hormones

These studies investigated the effects of neuropeptide Y (NPY) on in vitro release of luteinizing hormone-releasing hormone (LHRH) from the medial basal hypothalmus (MBH) and tested whether ovarian steroids modulate the LHRH response to NPY. Ovariectomized rats were implanted with 20-mm-long Silastic capsules containing a low concentration of estradiol (E2) (150 micrograms/ml oil), a high concentration of E2 (250 micrograms/ml oil), or sesame oil vehicle. Additional animals received high-dose E2 capsules plus an injection of progesterone (15 mg) concomitantly. Two days later, individual MBH fragments were incubated in medium alone for a 30-min period to obtain the basal rate of LHRH release, followed by a second 30-min period in medium containing NPY or saline. Exposure to NPY (10(-6) M) increased the release of LHRH from MBH of ovarian hormone-treated, but not from hormonally untreated rats. The LHRH response was most pronounced from the MBH of rats treated with either high-dose E2 or E2 plus progesterone. The increase in LHRH release was also elicited by 10(-7) M, but not by 10(-8) M NPY concentrations, using MBH from E2 plus progesterone-treated rats. In addition, NPY markedly potentiated the KCl-evoked release of LHRH from MBH of ovariectomized, hormonally untreated or low-dose E2-treated rats, under conditions when there was little or no effect of NPY on the basal LHRH release. Further, the release of LHRH stimulated by NPY was not accompanied by increase in the release of norepinephrine or of dopamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of neuropeptide Y, NPY analog (norleucine4-NPY), galanin and neuropeptide K on LH release in ovariectomized (ovx) and ovx estrogen, progesterone-treated rats☆

We studied the effects on plasma LH levels of intracerebroventricular (ICV) administration of neuropeptide Y (NPY), NPY analog (NPY-A), galanin (GAL) and neuropeptide K (NPK) in ovariectomized (ovx) and in ovx rats pretreated with estradiol benzoate (EB) and progesterone (P). Plasma LH levels were estimated in blood drawn from an intrajugular cannula before (0 min) and at 10, 20, 30 and 60 min after the ICV injection of either saline (3 microliter) or one of the neuropeptides in saline. The three classes of peptides elicited different LH responses in the two experimental paradigms. NPY and NPY-A (0.5 or 2 micrograms) decreased LH release in ovx rats and stimulated LH release in EBP ovx rats. However, GAL (0.5, 2 or 10 micrograms) failed to suppress LH release in ovx rats, but it readily increased plasma LH levels in a dose-related fashion in EBP ovx rats. In contrast, NPK readily decreased LH release in ovx rats in a time-related fashion for up to 60 min, but was mildly effective in EBP ovx rats as only a high dose of 10 micrograms produced a small significant increase. Collectively, our results show that (1) NPY can differentially effect LH release in ovx and EBP ovx rats but this property is not equally shared by the neuropeptides that have a similar anatomical disposition in the hypothalamus and (2) the excitatory effects of GAL are demonstrable in the steroid-primed rats and the inhibitory effects of NPK are apparent in the steroid-unprimed ovx rats. Since NPK induced a long-lasting marked suppression with little evidence of LH excitation at low doses, we speculate that either NPK alone or in conjunction with other peptides may mediate the suppression of LH release induced by gonadal steroids.

Evidence that neuropeptide Y is a physiological signal for normal food intake.

Neuropeptide Y is the most potent orexigenic signal known. To test the hypothesis that NPY is a physiological messenger molecule for normal food intake in rats, we studied the effects of passive immunization against endogenous NPY on cumulative daily food intake in non-fasted spontaneously feeding rats. The results show that continuous central infusion of NPY antibodies markedly suppressed the nighttime and the cumulative 24 h food intake in a dose-dependent fashion. These results support the hypothesis that NPY may be a physiological signal involved in the stimulation of ingestive behavior in rats.

Effects of naloxone on catecholamine and LHRH release from the perifused hypothalamus of the steroid-primed rat

The purpose of the present investigation was to determine the effects of naloxone on LHRH and catecholamine release from preoptic area-medial basal hypothalamic (POA-MBH) fragments in vitro. Ovariectomized rats were treated with estradiol benzoate, followed 2 days later by progesterone. The rats were killed 2 after progesterone administration and the POA-MBH dissected out and incubated in a perifusion system. After preincubation, medium with or without naloxone (1 mg/ml) was infused into the perifusion chambers and LHRH, norepinephrine, epinephrine and dopamine release were monitored. Naloxone concurrently released LHRH and the three catecholamines during the entire perifusion period. LHRH and catecholamine output returned to the control range within 30-45 min after cessation of naloxone infusion. These data show that naloxone can promptly stimulate catecholamine release and concur with the view that LH release evoked by naloxone in vivo may be due to hypersecretion of LHRH and they further raise the possibility that norepinephrine and epinephrine released in the vicinity of peptidergic neurons in the POA-MBH may participate in LHRH hypersecretion.

Effects of Ovarian Hormones on Norepinephrine and Dopamine Turnover in Individual Hypothalamic and Extrahypothalamic Nuclei

There is evidence that brain noradrenergic systems participate in the feedback regulation of luteinizing hormone (LH) and prolactin (PRL) secretion by ovarian hormones. The present study tested the hypothesis that administration of ovarian hormones in regimens that decrease LH release would also decrease the turnover of norepinephrine (NE), while ovarian hormone treatments that stimulate LH or PRL secretion would enhance the turnover NE in microdissected brain nuclei. Administration of estradiol to ovariectomized rats reduced circulating LH and decreased the turnover of NE in the periventricular nucleus. Treatment with estradiol, followed by progesterone or estradiol 72 h later enhanced the turnover of NE and of dopamine (DA) in the median eminence, and this was accompanied by elevations of LH and/or PRL. The dual estrogen treatment also increased NE turnover in the arcuate nucleus. Estradiol alone elevated DA turnover in the caudate nucleus and in the interstitial nucleus of the stria terminalis, and this was blocked by subsequent treatment with progesterone or estradiol. These findings indicate that activity in noradrenergic innervation to specific areas of the hypothalamus is altered by ovarian hormones. It is proposed that some of these changes may be involved in the feedback regulation of LH and PRL secretion by estradiol and progesterone.

Controlled Release of a Contraceptive Steroid from Biodegradable and Injectable Gel Formulations: In Vitro Evaluation

AbstractPurpose. The purpose of this study was to investigate the effects of formulation factors including varying wax concentration, drug loading and drug particle size, on drug release characteristics from both pure oil and gel formulations prepared with a combination of derivatized vegetable oil (Labrafil 1944 CS) and glyceryl palmitostearate (Precirol ATO 5), using levonorgestrel as a model drug.nMethods. The effects of varying drug loadings, different drug particle sizes, and wax (Precirol) concentrations on in-vitro drug release rates were evaluated, and the mechanisms of drug release from the gels were determined.nResults. Zero-order drug release rates from the 10% Precirol gel formulations containing 0.25, 0.50 and 2.00% w/v drug loadings were lower than those observed for oil formulations containing identical drug loadings. Higher zero-order release rates were observed from formulations containing smaller drug particles suspended in both oil and gel formulations. The mechanism of drug release from gels containing less than 0.25% w/w drug was diffusion-controlled. Increasing the wax concentrations in the gels from 5% w/w to 20% w/w significantly decreased the diffusivity of the drug through the gel formulations and markedly increased the force required to inject the gels from two different sizes of needles.nConclusions. This study shows how modification of the physicochemical properties of the gel formulations by changing the drug particle size, wax concentration and drug loading, affects drug release characteristics from the system.

Changes in pituitary oxytocin and vasopressin during the estrous cycle and after ovarian hormones: Evidence for mediation by norepinephrine

Abstract Pituitary levels of oxytocin and vasopressin were maximal on the morning of proestrus, declined during estrus, and were lowest on metestrus in cycling female rats. Norepinephrine levels in the paraventricular nucleus were decreased on proestrus and estrus when compared with metestrus-diestrus. Norepinephrine did not vary in the supraoptic nucleus. Administration of estradiol benzoate to ovariectomized rats elevated oxytocin in the pituitary 54 hr later. This elevation was not affected by a subsequent injection of estrogen or progesterone. Estrogen priming did not affect vasopressin levels in the pituitary, but a second injection of estrogen or of progesterone 48 hr later increased vasopressin in the pituitary when measured 6 hr after the second injection. Vasopressin was decreased 30 hr after a second injection of estrogen. The ovarian hormone treatments that elevated pituitary vasopressin decreased steady state levels of norepinephrine in the paraventricular nucleus and reduced the depletion of norepinephrine after administration of the catecholamine synthesis inhibitor α-methyltyrosine, suggesting a decrease in turnover. Ovarian hormones did not affect norepinephrine in the supraoptic nucleus. The present results suggest a role for posterior pituitary hormones in reproductive processes and a role for noradrenergic mechanisms in the paraventricular nucleus in mediating the effects of ovarian steroids on pituitary vasopressin.

Comparison of the effects of neuropeptide Y and adrenergic transmitters on LH release and food intake in male rats.

In view of the recent demonstrations that Neuropeptide Y (NPY) and adrenergic transmitters coexist in neurons of the rat brain, we have compared the effects of intraventricular (Ivt) injections of NPY and catecholamines on LH release and food intake in intact male rats. Of the three catecholamines, dopamine (DA), norepinephrine (NE) and epinephrine (E), only E (5.3 micrograms or 15.9 micrograms/rat) significantly stimulated LH release, although NE and E (5.3 micrograms/rat) were equally effective in eliciting food intake in satiated rats. Ivt administration of 10 micrograms NPY significantly stimulated LH release, whereas either lower (0.5 or 2 micrograms/rat) or higher (25 micrograms/rat) doses were ineffective. In contrast, NPY at doses of 0.5 - 10 micrograms/rat increased cumulative food intake in a dose-related fashion. These findings present preliminary evidence of the physiological correlates of the neuronal coexistence of adrenergic transmitters and NPY in the brain and raise the possibility that NPY may normally act either independently, in concert with or via adrenergic systems to evoke LH release and feeding responses in the rat.