Nori Geary

Cyclic estradiol treatment normalizes body weight and test meal size in ovariectomized rats.

We tested whether cyclic estradiol treatment, like continuous estradiol treatment, is sufficient to normalize meal size and body weight in ovariectomized rats. In Experiment 1, adult Long-Evans rats were ovariectomized and subcutaneously injected with 0, 0.2, or 2.0 microg estradiol benzoate (EB) in sesame oil each Tuesday and Wednesday. Oil-treated ovariectomized rats gained more weight during 4 weeks of ad lib feeding (48 +/- 5 g) than intact rats (16 +/- 1 g, p < 0.01). Cyclic treatment with 2.0 microg EB normalized weight gain (11 +/- 2 g). During the next week, plasma samples were assayed for estradiol. Cyclic treatment with 2.0 microg EB produced excursions of plasma estradiol that appeared similar to those of intact, cycling rats: estradiol level reached 190 +/- 60 pmol/L after the second EB injection before decreasing to undetectable levels (<30 pmol/L) by cycle end. In Experiment 2, test meal sizes after overnight food deprivation were measured. Cyclic treatment with 2.0 microg EB produced both tonic (i.e., at cycle onset, meal size was smaller in estradiol-treated than oil-treated rats) and phasic (i.e., meal size was smaller late in the EB-treatment cycle than early in it) decreases in meal size. Thus, a weekly cyclic regimen of estradiol treatment that produces changes in plasma estradiol concentration similar to those in intact cycling rats is sufficient to produce the body weight and meal size patterns that characterize normal hypothalamic-pituitary-gonadal function.

Pancreatic glucagon and postprandial satiety in the rat

The hypothesis that administration of pancreatic glucagon inhibits feeding by eliciting satiety for food was tested against several behavioral and physiological criteria of specificity. The effects of intraperitoneal glucagon injections on intake of a palatable milk diet were tested in rats maintained with ad lib access to pelleted diet. Injections of 25--800 micrograms/kg glucagon administered at meal onset inhibited meal size by 17--36%, but did not affect the normal postprandial behavioral satiety sequence or elicit any behavioral signs of toxicity. Latency to rest and intermeal interval were not affected. Glucagon decreased meal size by specifically inhibiting feeding during the terminal phase of the meal without affecting feeding earlier in the meal. This was also the case when glucagon was injected 4 min prior to meal onset. This range of glucagon doses did not affect water intake in water deprived rats consuming fluid volumes comparable to the milk intakes. They also did not affect body temperature. Finally, injection of 400 micrograms/kg glucagon after the initial exposure to a novel drinking fluid was not sufficient to form a conditioned taste aversion in a two bottle preference test. These data, together with reports that circulating pancreatic glucagon levels increase during meals, strongly suggest that pancreatic glucagon is involved in the production of postprandial satiety.

Cyclic estradiol replacement increases the satiety effect of CCK-8 in ovariectomized rats

The influence of cyclic ovarian hormone replacement therapy on the satiety effect of exogenous CCK-8 was determined to investigate the mechanism mediating the preestrous decrease in meal size in female rats. Once weekly, food-deprived ovariectomized rats were IP injected with 0.5-4 micrograms/kg CCK-8 and offered 0.4-0.8 M sucrose 52 h after the second of two daily SC injections of 2.5 or 10 micrograms estradiol benzoate or vehicle and 4 h after 500 mg progesterone or vehicle. In each of three tests, estradiol significantly increased CCK-8s inhibitory effect on sucrose intake. In contrast, progesterone alone or in combination with estradiol did not consistently influence the satiating potency of CCK-8. The interaction of estradiol and CCK-8 was clearest for the dose of 4 micrograms/kg CCK-8. The interaction occurred during diurnal tests and during dark-onset tests in which estradiol did not decrease baseline sucrose intake. These results demonstrate that a cyclic regimen of estradiol replacement in ovariectomized rats is sufficient to enhance the satiating effect of exogenous CCK-8 and that simultaneous progesterone treatment does not influence this effect. Potentiation of the satiating effect of CCK released from the small intestine by ingested food may be one of the mechanisms by which food intake decreases during the period of high estrogen concentration in the estrus cycle.

Hypothalamic implants of dilute estradiol fail to reduce feeding in ovariectomized rats.

To investigate further the site where estradiol (E(2)) inhibits food intake, we tested the effects on feeding of subcutaneous and intrahypothalamic implants of 10% E(2) benzoate in cholesterol (CHOL) or CHOL alone. E(2) was implanted subcutaneously in Silastic tubes, and intrahypothalamically via bilateral 29-gauge cannulas into the paraventricular nucleus (PVN) or the medial preoptic area (MPA) of ovariectomized (OVX) Sprague-Dawley and Long-Evans rats. Three-day implant periods followed 3-day baseline periods. Rats were allowed ad libitum access to chow and tap water, and food intake and body weight were measured each day. Subcutaneous 10% E(2) implants in Sprague-Dawley rats reduced food intake 21% on Day 2 and 34% on Day 3 (Ps<.01) and decreased 3-day body weight gain 11 g (P<.05). In contrast, 10% E(2) implants in the PVN of Sprague-Dawley rats did not change food intake or body weight. Implants of 10% or 20% E(2) in the MPA also failed to decrease food intake. MPA implants of 10% E(2) decreased body weight gain 8 g (P<.05), but MPA implants of 20% E(2) decreased weight gain only 4 g (P>.05). To determine whether the strain of rat affected our negative results on food intake, we implanted 10% E(2) into the PVN of Long-Evans rats. Again, PVN E(2) did not decrease food intake significantly in comparison to the pretest baseline. PVN E(2) did, however, decrease body weight gain 5 g and decreased food intake 6% compared to rats with implants of CHOL (both P<.05), but these effects appeared to be due to an increase in feeding in the CHOL group in comparison to their baseline. Finally, CHOL and E(2) implants did not impair the responsivity of the PVN because acute implants of norepinephrine (NE) into the PVN of E(2)- or CHOL-treated Long-Evans rats significantly increased food intake. Our results do not support the hypothesis that E(2)s actions in either the PVN or the MPA are sufficient to account for its inhibitory effects on feeding.

Overlapping Peptide Control of Alcohol Self-Administration and Feeding

This article represents the proceedings of a symposium at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL. The organizers and chairpersons were Mark Egli and Todd E. Thiele. The presentations were (1) Voluntary alcohol consumption is modulated by central melanocortin receptors, by Todd E. Thiele; (2) Central infusion of neuropeptide Y reduces alcohol drinking in alcohol-preferring P rats, by Robert B. Stewart and Nancy E. Badia-Elder; (3) The gut peptide cholecystokinin controls alcohol intake in Sardinian alcohol-preferring rats, by Nori Geary and Maurizio Massi; and (4) Hypothalamic galanin: a possible role in excess alcohol drinking, by Sarah F. Leibowitz and Bartley G. Hoebel.

Intracerebroventricular glucagon-like peptide-1 (7-36) amide inhibits sham feeding in rats without eliciting satiety

Glucagonlike peptide-1 (7-36) amide (GLP-1) and its receptors are present in several brain regions and may play a role in the physiological control of feeding. To investigate the effect of GLP-1 on eating in the absence of postingestive food stimuli, rats were implanted with gastric cannulas for sham feeding and lateral ventricular cannulas for infusion of GLP-1. Rats (n = 10) sham fed 0.8 mol/L sucrose for 45 min, beginning 5 min after intracerebroventricular (icv) infusion of 2.5 microL of artificial cerebrospinal fluid with 0-30 microg of GLP-1 . Behaviors were observed each minute using a time-sampling technique. Additionally, lick-by-lick records of the microstructural pattern of sucrose intake were made during the first 15 min of each test for five rats receiving 3 and 10 microg of GLP-1. GLP-1 decreased sham-fed intake by as much as 50%, but GLP-1 did not terminate sham feeding. The frequency of observations of feeding was decreased, but the frequency of resting, the terminal item in the behavioral sequence of postprandial satiety in real feeding rats, did not reliably increase. No abnormal behaviors were observed. Although GLP-I did not affect the latency to begin sham feeding, it significantly reduced the initial rate of licking. GLP-I did not affect the motor aspects of licking, because the interlick intervals within individual bursts of licking or overall lick efficiency were normal. These data suggest that intracerebroventricular infusions of GLP-1 inhibit sham feeding by decreasing the orosensory positive feedback that drives licking, rather than by activating physiological satiating mechanisms or nonspecific mechanisms such as aversion or motor incapacity.

Behaviorally specific inhibition of sham feeding by amylin

To further characterize amylins inhibitory action on feeding, we examined the effects of intraperitoneal injections of amylin on sham feeding of sucrose in food-deprived male rats with chronic gastric cannulas. Thirty and 100 microg/kg amylin reduced sham feeding, but did not terminate it or elicit the behavioral sequence of satiety. Real feeding of sucrose, but not sham feeding, was reduced after injection of 10 microg/kg amylin. Amylins inhibitory effect on sham feeding appeared behaviorally specific because neither 30 nor 100 microg/kg amylin affected sham drinking of water in thirsty rats and because no abnormal behaviors occurred. We conclude that amylin has a behaviorally specific satiating effect on sucrose sham feeding that is insufficient to elicit satiety in absence of gastric or postgastric food stimulation.

Ovariectomy and Estradiol Affect Postingestive Controls of Sucrose Licking

Ovariectomy (OVX) has been shown to increase, and estradiol replacement to decrease, meal size in rats. Because little is known about how estradiol influences meals, we conducted two experiments to examine the effects of OVX and beta-estradiol 3-benzoate (EB) replacement on the microstructure of licking behavior. In both experiments, patterns of licking were analyzed in adult female Sprague-Dawley rats during an 0.8 M sucrose test meal. In Experiment 1, meal microstructure was determined preOVX and 10-12 days postOVX. Rate of licking following OVX was not changed during min 1 of the meal, but was significantly faster during min 2-4 of the meal (p < 0.03). The numbers of bursts (runs of licks separated by 250-500 ms) and numbers of clusters (runs of licks separated by > 500 ms) were significantly increased during min 2-4 (p < 0.05). In Experiment 2, OVX rats received EB replacement. Rate of licking after EB replacement was not changed during min 1 of the meal, but was significantly slower during the remainder of the meal (min 2-4, min 5-7, and min 8-10). Burst size, cluster size, and interburst interval were less after EB replacement during min 5-7 of the test meal (all p < 0.05). Because both OVX and EB replacement failed to alter the rate of licking during min 1, estrogen did not appear to alter the palatability of sucrose. OVX and EB replacement did appear to affect a postingestive mechanism(s) that is engaged within 2-4 min of meal onset.

The increased satiating potency of CCK-8 by estradiol is not mediated by upregulation of NTS CCK receptors

Estradiol benzoate (EB) increases the satiating effect of CCK-8 in ovariectomized rats. It is possible that this effect of EB is due to upregulation of CCKA receptors in the terminals of vagal afferent fibers because these receptors have been implicated in the mediation of the satiating effect of intraperitoneally injected CCK-8. To test this hypothesis, we used in vitro quantitative autoradiography to measure the effects of EB on the binding characteristics of CCK receptors in the nucleus tractus solitarius (NTS), a region that contains central terminal projections of abdominal vagal afferent fibers. As additional measures of EBs effects on CCK receptors, we also characterized EBs effects on CCK.8 binding in the area postrema (AP), a brain region rich in CCKA receptors, the ventromedial hypothalamus (VMH), a region rich in CCKB receptors, and in the pancreas, a gland rich in CCKA receptors. Saturation binding experiments were run using [125I]CCK-8 (approximately 40 pM, 2200 Ci/mmol) and 0.1-100 nM unlabelled CCK-8. EB did not change the number (Bmax) or affinity (Kd) of CCK receptors in the NTS. Furthermore, competition experiments with 500 nM of the selective CCKA receptor antagonist devazepide or the selective CCKB antagonist L365,260 demonstrated that EB failed to affect CCK receptor subtype number in the medial and lateral divisions of the NTS. EB also did not affect binding in the AP or VMH. These results do not confirm our hypothesis. The lack of effect of EB on vagal CCKA receptors in the NTS was not due to inappropriate conditions of tissue sampling or autoradiographic technique because EB increased the number, but not the affinity, of CCKA receptors in the pancreas significantly.

Brief, meal-contingent infusions of gastrin-releasing peptide1-27 and neuromedin B-10 inhibit spontaneous feeding in rats.

We have previously shown that brief, meal-contingent infusions of the amphibian peptide bombesin (BN) reduce meal size in spontaneously feeding rats. These inhibitory effects presumably reflect actions of BN on receptors for its mammalian homologues, which include gastrin-releasing peptide1-27 (GRP) and neuromedin B-10 (NMB). In the present study, we used the spontaneous feeding paradigm to explore the effects of meal-contingent infusions of these mammalian homologues. Undisturbed, ad lib-fed male rats (n = 12) with chronic inferior vena caval catheters were infused with saline, 5 nmol/kg GRP, 5 nmol/kg NMB, and a mixture of 5 nmol/kg GRP and 5 nmol/kg NMB in counterbalanced order, with intervening noninfusion days. Infusions were remotely activated at the onset of the first nocturnal meal and continued for 3 min (34 microliters/min), delivery being completed within the first 2 min of infusion. Feeding was measured via electronic balances linked to a computer. The effects of all peptide conditions were confined to the first nocturnal meal. Significant and comparable decreases in the size and duration of this meal were observed with GRP, NMB, and GRP + NMB, with no significant effects on the intermeal interval or satiety ratio (intermeal interval/meal size). Thus, brief vena caval infusions of GRP and NMB, given alone or together at the onset of the first nocturnal meal, significantly reduced meal size and duration in spontaneously feeding rats.