Phyllis Callahan

Orphanin FQ stimulates prolactin and growth hormone release in male and female rats

Intracerebroventricular administration of Orphanin FQ (5.5, 55 or 550 pmol) caused a dose-related increase in prolactin secretion in both male and female rats and stimulated GH secretion in males. The magnitude of the prolactin secretory response was greater in females than in males. These effects of OFQ on prolactin and growth hormone release are the same as the stimulatory effects of the endogenous opioid peptides.

INHIBITION OF TUBEROINFUNDIBULAR DOPAMINERGIC NEURAL ACTIVITY DURING SUCKLING : INVOLVEMENT OF MU AND KAPPA OPIATE RECEPTOR SUBTYPES

Previous studies have shown that mu (μ) and kappa (κ) opioid antagonists inhibit suckling‐induced prolactin release. Prolactin responses elicited by pup suckling or opioid administration are mediated, at least in part, by suppression of dopamine (DA) release from tuberoinfundibular dopaminergic (TIDA) neurons in the hypothalamus. We examined the effects of the μ opiate receptor antagonist, β‐funaltrexamine (β‐FNA), and the κ opiate receptor antagonist, nor‐binaltorphimine (nor‐BNI) on the activity of TIDA neurons in lactating rats. TIDA neuronal activity was determined by measuring DOPA accumulation in the caudate putamen (CP) and median eminence (ME). The effects of opioid antagonist treatment were determined in pup‐deprived (low circulating prolactin levels) or pup‐suckled rats (high circulating prolactin levels). The accumulation of 5‐hydroxytryptophan (5‐HTP) in the medial preoptic area (MPOA), the anterior hypothalamus (AH) and the median eminence (ME) was quantified as an index of serotonergic activity in the same animals for comparative purposes.

Exercise improves biomarkers of health and stress in animals fed ad libitum

Voluntary and forced exercise decrease morbidity and mortality in laboratory animals. Caloric restriction has similar effects on health and unique benefits on life span. Nonetheless, in most experiments, animals do not have access to physical activity and are fed ad libitum (AL). We hypothesized that with regular access to either unlimited running wheel exercise (EX) or limited physical activity (PA), key biomarkers of health would be enhanced enough to counter some consequences of a sedentary AL lifestyle. This 16-month study compared body weight, tumor number and size, tissue lesions, oxidative stress, and reactive stress in (1) sedentary animals with no access to physical activity (SED); (2) animals with access to hour-long, twice weekly activity in a large box (PA); and (3) animals with access every other day to a running wheel (EX). At the end of the study, EX body weight was 8-9% lower than PA and SED. In addition, EX had no kidney lesions versus 50% in PA and SED, and had smaller tumor size (10+/-2 vs. 14+/-4 and 30+/-4 mm). Exhaustive exercise lowered glutathione/oxidized glutathione ratio in EX and PA, but in SED, the ratio was depressed even in resting animals. In all treatments, prolactin (PRL) levels were lower in resting animals than in acutely exercised animals. In conclusion, EX had the most favorable health biomarkers while SED had the least. PA did not confer gross health benefits different than the SED group, but was biochemically more similar to EX animals.

Immunoneutralization of Endogenous Opioid Peptides Prevents the Suckling-Induced Prolactin Increase and the Inhibition of Tuberoinfundibular Dopaminergic Neurons

Previous studies have shown that the endogenous opioid peptides, acting at specific opiate receptor subtypes, are involved in the suckling-induced prolactin secretory response. The prolactin increase elicited by suckling is due, at least in part, to an inhibition of tuberoinfundibular dopaminergic (TIDA) neurons in the hypothalamus. We investigated the effects of immunoneutralization of dynorphin, leu-enkephalin and met-enkephalin on the suckling-induced prolactin increase and on the activity of the TIDA neurons in lactating female rats between days 7 and 12 postpartum. Rats were injected into the right lateral ventricle with antiserum specific for one of these three peptides. Control rats were administered equal amounts of immunoglobulin proteins. Suckling produced a profound and significant increase in prolactin levels, as well as a decrease in DOPA accumulation in the median eminence of lactating rats. Administration of immunoglobulin concentrations of up to 3.6 µg did not inhibit the prolactin secretory response to the suckling stimulus and did not prevent the suckling-induced inhibition of TIDA neurons. Antisera to all three endogenous opioid peptides abolished the suckling-induced prolactin increase and prevented the inhibition in DOPA accumulation in the median eminence. Thus, the endogenous opioid peptides, dynorphin, leu-enkephalin and met-enkephalin, are essential for the prolactin secretory response to suckling and inhibition of TIDA neuronal activity is at least one of the mechanisms of action utilized by these peptides.

Estrogen regulation of neurotrophin expression in sympathetic neurons and vascular targets

We hypothesize that estrogen exerts a modulatory effect on sympathetic neurons to reduce neural cardiovascular tone and that these effects are modulated by nerve growth factor (NGF), a neurotrophin that regulates sympathetic neuron survival and maintenance. We examined the effects of estrogen on NGF and tyrosine hydroxylase (TH) protein content in specific vascular targets. Ovariectomized, adult Sprague-Dawley rats were implanted with placebo or 17beta-estradiol (release rate, 0.05 mg/day). Fourteen days later, NGF levels in the superior cervical ganglia (SCG) and its targets, the heart, external carotid artery, and the extracerebral blood vessels, as well as estrogen receptor alpha (ERalpha) content levels in the heart, were determined using semi-quantitative Western blot analysis. TH levels in the SCG and extracerebral blood vessels were determined by Western blotting and immunocytochemistry, respectively. Circulating levels of 17beta-estradiol and prolactin (PRL) were quantified by RIA. Estrogen replacement significantly decreased NGF protein in the SCG and its targets, the external carotid artery, heart and extracerebral blood vessels. TH protein associated with the extracerebral blood vessels was also significantly decreased, but ERalpha levels were significantly increased in the heart following estrogen replacement. These results indicate that estrogen reduces NGF protein content in sympathetic vascular targets, which may lead to decreased sympathetic innervations to these targets, and therefore reduced sympathetic regulation. In addition, the estrogen-induced increase in ERalpha levels in the heart, a target tissue of the SCG, suggests that estrogen may sensitize the heart to further estrogen modulation, and possibly increase vasodilation of the coronary vasculature.

Opiate Receptor Subtype Involvement in the Stimulation of Prolactin Release by β-Endorphin in Female Rats

The prolactin secretory response to beta-endorphin and the involvement of opiate receptor subtypes in this response was determined in both diestrous and postpartum, lactating female rats. The involvement of the mu-, delta- and/or kappa-site was determined by administering specific antagonists for each of these sites prior to beta-endorphin. beta-Funaltrexamine (beta-FNA, 1 or 5 micrograms) was administered to block mu-sites, ICI 154,129 (5, 10 or 25 micrograms) blocked delta-sites and nor-binaltorphimine (norBNI, 8 micrograms) blocked kappa-sites. The ability of beta-FNA and ICI 154,129 to block prolactin secretion following morphine administration was also determined. A dose response study for beta-endorphin indicated that beta-endorphin, at doses as low as 25 ng, was a potent stimulus for prolactin release producing an increase in prolactin that mimicked the suckling-induced prolactin increase. In addition, all three antagonists were capable of antagonizing the stimulatory effect of beta-endorphin in both diestrous and postpartum female rats. These results indicate that beta-endorphin is a potent stimulus for prolactin secretion and that these three opiate receptor subtypes interact to produce its stimulatory effect on prolactin release.

Effects of immobilization stress on tuberoinfundibular dopaminergic (tida) neuronal activity and prolactin levels in lactating and non-lactating female rats

The effects of immobilization stress on the prolactin secretory response and on the activity of the tuberoinfundibular dopaminergic (TIDA) neurons were determined in intact, virgin female rats on the morning of diestrus or proestrus and in post-partum, lactating female rats. The virgin females exhibited a significant increase in circulating levels of prolactin which was evident by 1 minute and persisted during the immobilization (5 minutes). In contrast, the prolactin secretory response in lactating females was significantly attenuated compared to non-lactating animals. The activity of the TIDA neurons was not altered by the 5 minutes of stress. Even after 30 minutes of immobilization, TIDA neuronal activity was not affected in either the lactating or cycling females. These data suggest that the cycling female rat is capable of a prolactin secretory response to the stressor without inhibition of TIDA neuronal activity. It seems likely that prolactin releasing factors mediate this response. In contrast, stress did not produce a similar prolactin increase during lactation. It seems likely that, during lactation, the pituitary is not sensitive to releasing factors unless the TIDA neurons are inhibited. There appear to be differences in the sensitivity of the pituitary depending on the physiological state of the model employed.

Cadmium exposure inhibits the prolactin secretory response to thyrotrophin releasing hormone (TRH) in vitro

In vitro dose response (0.1-100 microM) and time course studies were performed on anterior pituitary cells isolated from female Sprague-Dawley rats in the diestrous stage of the estrous cycle. All doses of TRH produced a significant increase in prolactin release by 5 min which was sustained for 30 min. Pretreatment with 25 or 50 microM Cd for 2 or 4 h produced a significant decrease in prolactin secretion during Cd exposure and in the prolactin secretory response to 1 microM TRH. These results suggest that Cd pretreatment alters the subsequent ability of the lactotrophs to respond to the physiological secretagogue TRH. This alteration does not appear to be due to a rebound phenomenon since basal prolactin release was the same in Cd-pretreated and control cells once the metal was removed. These results suggest that Cd pretreatment produces a significant decrease in prolactin release from the anterior pituitary lactotrophs in response to subsequent TRH stimulation in vitro.

Frogs reabsorb glucose from urinary bladder

The amphibian urinary bladder is a bilobate, highly distensible and vascularized sac that stores fluid for use during periods of water stress. The organ is composed of a thin basement membrane overlaid by a selectively permeable ‘tight’ epithelium and is important in the homeostatic regulation of ion and osmolyte balance. We report here that glucose reabsorption from the urinary bladder permits recovery of sugar destined for excretion in the freeze-tolerant frog Rana sylvatica, whose unique winter survival strategy invokes extreme hyperglycaemia and ultimately, glucosuria.

IMMUNONEUTRALIZATION OF β-ENDORPHIN BLOCKS PROLACTIN RELEASE DURING SUCKLING WITHOUT AFFECTING TUBEROINFUNDIBULAR DOPAMINERGIC NEURAL ACTIVITY

The effect of immunoneutralization of beta-endorphin on the suckling-induced prolactin increase and on the activity of the tuberoinfundibular dopaminergic (TIDA) neurons was determined in lactating female rats between days 8 - 12 post-partum. Two antisera were used in the immunoneutralization studies. Both were specific for beta-endorphin, exhibiting little cross reactivity with met- or leu-enkephalin or dynorphin. Antisera to beta-endorphin completely abolished the suckling-induced prolactin increase indicating that this endogenous opioid peptide is involved in this response. Suckling significantly inhibited DOPA accumulation in the median eminence and antiserum to beta-endorphin did not prevent this inhibition. Additionally, 5-endorphin antiserum significantly reduced TIDA neural activity even in pup-deprived dams. These results indicate that beta-endorphin is involved in the prolactin secretory response to suckling but that inhibition of TIDA neuronal activity is not its mechanism of action. Other possible mechanisms are discussed.