Development of food motivated behavior in free feeding and food restricted zucker fatty (fa/fa) rats

The food motivated behavior of ad lib fed or calorically restricted male Zucker fatty and lean rats was compared at 12, 16, 20 and 30 weeks of age. The groups were fasted for 0, 12 and 24 hr and required to lever press for food pellets on VI 40 sec. Responding by ad lib fatties was elevated over lean controls during the rapid growth period (12 and 16 weeks of age) but was significantly reduced following the completion of growth (20 weeks). Elevated responding accompanied hyperphagia by the ad lib fatties during growth but did not accompany a second, adult onset period of hyperphagia in these fatties. In contrast, fatties calorically restricted from birth responded at elevated rates at all ages compared to lean controls. At 20 weeks restricted fatties, which were as obese on a percentage body composition basis as ad lib fatties, responded significantly more than the latter group at all fasting levels. Metabolic measurements revealed elevated fat cell size, LPL activity/cell and insulin levels in both fatty groups at 15 weeks, and elevated fat cell number in these groups at 33 weeks. The possibility is discussed that enhanced lipogenic factors present during the rapid growth period combine with the metabolic demands of growth to produce a “compensatory hyperphagia” in the fatty rat.     

Ejaculatory and postejaculatory behavior of male and female rats: effects of sex hormones and electric shock

Ejaculatory patterns were observed in normally reared, postpuberally castrated male and female rats treated with sex hormones and electrical shock in adulthood. In a preliminary experiment, 3 females treated with 50 microgram estradiol benzoate (EB) and 500 microgram progesterone (P) showed intromission and ejaculatory behavior when subjected to shock. Refractory periods were abnormally short and there was almost no postejaculatory vocalization. In Experiment 1, males and females were tested with electrical shock following daily treatment with 32 microgram EB with and without 500 microgram P on the test day. There was no difference between males and females in preejaculatory behavior, but females displayed abbreviated refractory periods and no postejaculatory vocalization. Progesterone had no observed effect. In Experiment 2 castrated males and females were subjected to shock after treatment with 8 microgram EB per day for 3 weeks. No P was given. Again females showed drastically reduced refractory periods and little vocalization. In Experiment 3, males and females treated with TP and shock displayed ejaculatory patterns, normal refractory periods and vocalization. Results show that female rats are capable of exhibiting the ejaculatory response without sex hormone treatment in perinatal life or androgen treatment in adulthood. It was also demonstrated that there is a sex difference in the postejaculatory behavior shown by estrogen-treated male and female rats.     

Food intake and body weight modifications following medial hypothalamic hormone implants in female rats

Two groups of female rats had crystalline estradiol benzoate, progesterone and cholesterol implanted in the medial hypothalamus. One group was ovariectomized (OVX) and weighed approximately 300 g. The second group was adrenalectomized and ovariectomized (ADX-OVX) and weighed approximately 250 g. EB placed in the ventromedial hypothalamus significantly reduced feeding in the OVX animals; progesterone and cholesterol had no effect. In contrast, progesterone increased feeding in the lean ADX-OVX animals; EB and cholesterol were without effect. The most effective progesterone placements were in the dorsal medial hypothalamic nuclei. The data are discussed in terms of EB and progesterone influencing a hypothalamic set point for body weight.     

Onset of estrogen replacement has a critical effect on synaptic density of CA1 hippocampus in ovariectomized adult rats

OBJECTIVES: The aim of this study was to evaluate differences between estrogen replacement therapy initiated either 4 or 12 days after ovariectomy on the synaptic density of the hippocampal CA1 field in rats. DESIGN: Female, adult, Wistar rats were ovariectomized bilaterally under ether anesthesia and divided among the following groups: 1) estrogen (conjugated equine estrogen 50 microg in 0.5 mL of propylene glycol, daily, p.o. gavage, for 60 days), starting 4 days after ovariectomy (n = 5); 2) propylene glycol (0.5 mL daily, p.o. gavage, for 60 days), starting 4 days after ovariectomy (n = 4); 3) estrogen (conjugated equine estrogen 50 microg in 0.5 mL of propylene glycol, daily, p.o. gavage, for 45 days), starting 12 days after ovariectomy (n = 3); 4) propylene glycol (0.5 mL daily, p.o. gavage, for 45 days), starting 12 days after ovariectomy (n = 3). At the end of the treatment, the rats were processed for electron microscopy and light analysis. RESULTS: Synaptic density in all of the CA1 strata subjected to evaluation was significantly higher in animals in which estrogen replacement was initiated 4 days after ovariectomy as compared with controls. In contrast, initiation of treatment after a 12-day interval did not result in recovery of synaptic density in any of the CA1 strata and was significantly lower than that of the animals subjected to hormone replacement after a 4-day delay (P < 0.01). CONCLUSION: The delay for hormone replacement therapy might have critical implications for modulating synaptic density.     

Cholecystokinin stimulates and inhibits lordosis behavior in female rats

Recently, IP CCK-8 has been shown to inhibit lordosis in sexually experienced, estradiol benzoate (EB) and progesterone (P) primed rats. However, receptivity is influenced by prior sexual experience and/or exposure to sex steroids, as well as the steroid dosage administered before testing. Thus, we examined the effect of CCK-8 (3 micrograms/kg; IP) on lordosis in rats with different degrees of receptivity. Three weeks after ovariectomy, females were treated with EB followed 48 hr later with P, or with EB alone. CCK-8 significantly facilitated lordosis in rats given 5 micrograms EB. Following a 5 week nonexperimental period, animals were more receptive and CCK-8 significantly inhibited lordosis in the 5 or 10 micrograms EB groups. In a separate experiment, rats were ovariectomized, adrenalectomized, and treated with EB alone. As in the first experiment, CCK-8 facilitated and inhibited lordosis. CCK-8's effects were highly dependent on the female's receptivity, facilitating lordosis when receptivity was low and inhibiting lordosis when receptivity was high (but not maximal). In conclusion, IP CCK-8 modulates lordosis behavior independent of P, but its effects depend on the female's degree of receptivity.     

Hyperinsulinemia and its role in maintaining the hypothalamic hyperphagia in chickens

Body weights and skeletal growth of female rats treated neonatally with low doses of testosterone propionate (TP) or estradiol benzoate (FB) were greater than oil-treated controls. After ovariectomy at 75 days of age EB-treated animals gained less weight than did the oil-treated controls and TP-treated rats which were comparable in weight gain. Neonatal treatment with TP or EB produced decreased sensitivity to the anorexic and weight-limiting effects of estrogen treatment after ovariectomy. However, all groups were equally sensitive to the anorexic effects of a single dose of CI-628. The possible mechanisms by which neonatal treatments with gonadal hormones influence food intake and body weight regulation are discussed.     

Early life handling decreases serotonin turnover in the nucleus accumbens and affects feeding behavior of adult rats

In our previous studies, we reported that neonatally handled rats have an increased ingestion of sweet food but are resistant to the damaging effects of a chronic exposure to a highly palatable diet. Accumbal serotonin (5‐HT) is important for feeding behavior and plays a role in the vulnerability to diet‐induced obesity. Therefore, our hypotheses were (1) 5‐HT turnover in the nucleus accumbens is altered in neonatally handled animals and plays a role in their differential feeding behavior and (2) if this is so, a chronic pharmacological treatment affecting 5‐HT reuptake (chronic imipramine) would be able to revert the behavioral findings. Litters were divided into nonhandled and handled (10 min/day, Days 1–10 after birth). In Experiment 1, we demonstrated that a decreased 5‐HT metabolism in the nucleus accumbens was observed in adult handled animals. In Experiment 2, the two previous groups were subdivided and assigned to receive imipramine diluted in water or water alone. After 30 days of treatment, we evaluated their weight gain and feeding behavior. Handled rats weighed less than nonhandled rats, and all imipramine‐treated rats showed a reduction in weight gain after 60 days of treatment. Imipramine reverted the increased sweet food consumption seen in neonatally handled rats. We conclude that serotonin is involved in the altered feeding behavior of neonatally handled rats, and this protocol is an important tool for studying the mechanisms by which early life events have a long‐term impact on feeding preferences. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 52:190–196, 2010     

Effect of estradiol and progesterone on daily rhythm in food intake and feeding patterns in Fischer rats

The product of meal number x meal size, over time, is food intake. Because estrogens modulate feeding activity via their action on the hypothalamus, and because there is a diurnal rhythm in the expression of cytoplasmic estrogen receptors and in estrogen binding activity, the present study examined the effects of ovariectomy and later hormone therapy on acute changes in body weight, and on the meal number-to-meal size relationship as reflected by food intake in the dark/light feeding patterns, in adult female rats in the intact state and after ovariectomy. Twelve female Fischer rats were randomized into ovariectomy and sham operation groups. A rat eater meter measured the feeding indexes for 15 days before and 25 days after ovariectomy, and later for 35 days with hormone therapy. We report: (a) mean body weight gain was linear before and up to ovariectomy, while exponential after ovariectomy; (b) increase in daily food consumption is mainly via an increase in food intake during the light phase; (c) light phase meal number remains unchanged, meal size significantly increases, with the resultant increase in overall food intake; (d) during the dark phase, meal size also significantly increases, but is accompanied by a proportional decrease in meal number, resulting in unchanged dark-phase food intake; and (e) estrogen restoration with either estradiol valerate or estradiol-progesterone combination, reversed the above changes. Data show that in the female Fischer 344 rat: (a) changes in daily rhythm in food intake are brought about by differential effects of the hormones on both meal size and meal number in both the total daily levels as well as in the dark-to-light distribution; (b) estadiol appears to have a tonic inhibitory effect on the light phase meal size and a phasic effect on the dark phase meal size and number, but no significant effect on the light-phase meal number; and (c) in the Fischer rats, progesterone augments estradiol's effect on these indicies.     

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.     

Pelvic and pudendal nerves influence the display of paced mating behavior in response to estrogen and progesterone in the female rat.

Two experiments examined whether the pudendal and pelvic autonomic nerves are important for pacing behavior shown by the female rat during mating. Ovariectomized female rats received bilateral transection of the pudendal (Pux), pelvic (Pex), or pudendal and pelvic (Pu + Pex) nerves or were sham-operated (Shx). Lordotic behavior, precopulatory solicitations, postural adjustments, and pacing behavior were measured 14 days after nerve transection in Experiment 1 after sequential treatment with estradiol benzoate (EB) and progesterone (P) and in Experiment 2 on the day after 7 daily injections of EB. After combined EB and P treatment, disruption of pacing behavior occurred in Pex and Pu + Pex animals. After EB-only treatment, Pux animals as well as Pex and Pu + Pex animals showed decreased pacing behavior. Thus, afferent input via the pelvic nerve is important for the display of pacing behavior, and P may counteract the effects of autonomic nerve transection.     

Importance of feeding time in pair-fed, ovariectomized rats

The present investigation was designed to determine the effects of feeding time on body weight gain in ovariectomized (OVX) rats pair-fed with sham operated control rats. In pair feeding, a test and a control rat were each fed the same amount of food. In the first experiment, OVX rats and controls were pair-fed at 1900, just before the dark phase of the light-dark cycle. Body weight gain was observed for 36 days. There was no significant difference in weight gain between the two groups. In the second experiment, when rats were pair-fed chow, starting from 1200, there was a significant increase in the body weight of the OVX group compared to the sham operated control group. Observation continued for 56 days. In the third experiment, the OVX group in the pair-fed condition and the sham control group were pair-fed at 1900 for 56 days and feeding was then switched to 0500 (just before the beginning of the light cycle) for 28 days. The body weight gain in the OVX pair-fed group was quite similar to that of the sham control group when feeding started at 1900, but a significant increase in body weight gain of the OVX group was observed after switching the starting time of feeding to 0500. In conclusion, the results suggest that the time of daily feeding may be important for body weight gain during the dynamic phase after ovariectomy.     

Sexual behavior of male and female septal lesioned rats

Gonadectomized male and female rats were given septal lesions (SL) or sham surgery at approximately 60 days of age. After 3 weeks lordosis behavior tests were initiated. Females were tested after daily injections of 2 μg estradiol benzoate (EB) for 3 days, while males were tested after EB only (2 μg×3 days), and after EB plus progesterone (Prog). The mean lordosis quotients (LQ) of septal lesioned female rats were significantly higher than those of sham operated controls. No increase in lordosis responding was seen in male rats with either EB alone or EB+Prog. Following an additional 3 week interval without steroid treatment masculine behavior tests began. All animals received a pretest and were tested again on Day 4, 7, 11 and 15 daily tesosterone propionate (150 μg/day) treatment. No alterations in masculine sexual behavior (relative to that of controls) were found in either male or female septal lesioned rats. It is concluded that the increased hormone sensitivity is specific for lordosis behavior, at least when the SL are given in adulthood.     

Dietary effects upon food and water intake and responsiveness to estrogen, 2-deoxy-glucose and glucose in female rats

Female rats were maintained on three different diets—chow (C), high-dextrose (D) and high-fat (F) diets—and effects upon feeding of ovariectomy, estradiol benzoate (EB), 2-deoxy-glucose (2-DG) and ad lib glucose solutions were followed. F-fed rats responded significantly more to EB, 2-DG and to ad lib glucose than did C- or D-fed rats; increased responses to EB were not explainable on the basis of increased body weight (BWt). D-fed rats chronically maintained a lowered caloric intake, water intake, and BWt, in spite of a high palatability of the D-diet. Responses of D-fed rats to EB showed a high correlation with BWt, suggesting an augmented influence of a BWt-related signal upon feeding regulation in D-fed rats. A possible relationship between carbohydrate metabolism and responsiveness to EB is discussed.     

Dietary self-selection in intact, ovariectomized, and estradiol-treated female rats

The effects of estrogenic stimulation on diet selection were examined in intact, estrous cycling rats, ovariectomized (OVX) rats, and OVX rats given estradiol benzoate (EB) hormone replacement therapy. In Experiment 1, OVX was associated with the nearly exclusive choice of the more calorically dense diet of a pair of diets varying in the concentration of one of the three basic macronutrients (i.e., fat, carbohydrate, and protein), an effect that was decreased by EB administration. In the second experiment, dietary self-selection was examined in intact, estrous cycling rats given access to an isocaloric diet triplet of fat, carbohydrate (CHO), and protein. Total caloric intake and body weight did not vary across the estrous cycle. However, diet selection did vary. Fat intake increased; CHO and, to a lesser extent, protein intake decreased during estrus. An opposite diet selection occurred during diestrus. In Experiment 3, OVX resulted in progressive increases in CHO and protein intake, with a concurrent decrease in fat consumption. The EB treatment partially reversed this diet selection profile (Experiment 4). These results were confirmed by diet pairs with both naturally occurring and experimentally produced estrogenic stimulation (Experiments 5 and 6). These data are consistent with the findings of previous research demonstrating estrogenic reduction in CHO intake with standard high-CHO commercial diets. In addition, an increase in fat intake during estrogenic stimulation was found.     

Effects of estrogen upon feeding inhibition produced by intragastric glucose loads

Two studies were performed to determine if estrogen could potentiate the inhibitory effects of glucose on feeding behavior. In the first experiment, ovariectomized rats were injected with either 6 μg of estradiol benzoate (EB) or with sesame oil, and two days later were given 5 ml intragastric loads of either 40% glucose or 13.5% urea by gavage. Gavage was performed after a 12-hr fast, during the light period. Food intake (FI) was measured at hourly intervals, starting 2 hr after gavage and reaccess to food as well as daily throughout the experiment. In a second experiment, rats received a similar treatment, except that gavage was performed in an unfasted condition, during the dark period. It was found that (1) a 12-hr fast abolishes anorexic effects of EB, but leaves the inhibitory effects of glucose intact, (2) EB does not potentiate short-term effects of glucose, and (3) EB enhances delayed (22 hr after gavage) effects of glucose upon feeding. These findings are consistent with the hypothesis that EB modified the long-term control of feeding behavior, probably at the level of the ventromedial hypothalamus.     

Thyroxine treatment reduces the anorectic effect of estradiol in rats

In order to examine a possible interaction between thyroxine and estradiol in the control of feeding, 14 ovariectomized adult female rats were given daily injections of 9.8 micrograms/100 g of body weight of 1-thyroxine (T). Another 14 rats received 0.15 ml of saline (S) subcutaneously each day, and food intake was measured for both groups daily. After 15 days of treatment, 8 rats from each group were also given a single injection of 6 micrograms of estradiol benzoate (EB), and the remaining 6 rats of each group received peanut oil vehicle. It was found that T-treated rats showed a significantly smaller drop in food intake after EB than did S-treated rats. This thyroxine-induced decrease in responsiveness to EB did not appear due to any changes in body weight per se but may be related to effects of thyroxine on general metabolism.     

Chronic infusion of the amylin antagonist AC 187 increases feeding in Zucker fa/fa rats but not in lean controls

Numerous studies have established the pancreatic B-cell hormone amylin as an important anorectic peptide affecting meal-ending satiety. In the present study, we investigated the effect of a chronic infusion of the amylin antagonist AC 187 on food intake. The studies were performed using obese Zucker fa/fa rats, which are hyperamylinemic but have a defective leptin and insulin signaling system. A chronic intraperitoneal infusion of the amylin antagonist AC 187 (10 microg/kg/h) significantly increased dark phase and total food intake in Zucker but not in lean control rats. During the 8-day infusion experiment, AC 187 had no clear effect on body weight gain in either group. After acute administration, amylin and its agonist salmon calcitonin (sCT) equally reduced food intake in Zucker and lean control rats while cholecystokinin's (CCK) anorectic effect was weaker in the Zucker rats. We provide evidence for amylin being a potential long-term regulator of food intake because AC 187 increased food intake in obese fa/fa rats but not in lean control animals, which have low baseline amylin levels. Amylin may play some role as lipostatic feedback signal similar to leptin and insulin at least when the leptin and insulin feedback signaling systems are deficient. Despite basal hyperamylinemia in the Zucker rats, they do not seem to be less sensitive to the anorectic effects of amylin or its agonist sCT than respective controls. This contrasts with CCK whose anorectic action is reduced in Zucker rats when compared with lean controls.     

Androgen inhibition of sexual receptivity is modulated by estrogen

Sexual receptivity induced in ovariectomized rats by the long-term administration of estradiol benzoate (EB) can be inhibited by concurrent administration of androgens. Experiment 1 examined the role of time course and dose of androgens in the inhibition of estrogen-induced sexual receptivity. Ovariectomized rats were treated with EB (2.0 microg per rat per day) for 6 days and tested for sexual receptivity (Test Day I). EB treatment continued for 15 days concomitant with daily administration of one of three doses of dihydrotestosterone propionate (DHTP; 7.5, 0.75, 0.075 mg/kg) or 3α-androstanediol (3α-Adiol; 3.75, 1.0, 0.375 mg/kg). Four tests for sexual receptivity were conducted on days 3, 6, 14, and 15 of the androgen/vehicle treatment period (Test Days II-V). On Day 15 (Test Day V), the rats received progesterone (1.0 mg per rat) 4 h before testing. Using the same experimental design, Experiment 2 examined the effect of increasing the dose of estrogen on the androgenic inhibition of sexual receptivity. Ovariectomized rats were treated with one of two doses of EB (2.0 or 10.0 microg per rat per day) concomitant with daily administration of DHTP (7.5 mg/kg) or 3α-Adiol (3.75 mg/kg). In Experiment 1, the highest doses of both DHTP and 3α-Adiol significantly inhibited estrogen-induced sexual receptivity. Data from Experiment 2 indicate that the inhibitory effects of DHTP but not 3α-Adiol can be moderated by an increased dose of EB.     

Attractivity of male rats induced by estradiol and progesterone

From the present study it appears that castrated male rats can be made attractive to intact males by estradiol benzoate plus progesterone (EB + P). For the determination of attractivity a residential plus-maze was used in which the resident male could choose between the company of the experimental male and that of an ovariectomized female with or without EB + P. In one experiment the males were castrated in adulthood; these animals were less attractive than the ovariectomized females with a similar hormone treatment. In a second experiment it was found that males that had been castrated at birth were as attractive as the ovariectomized females, after treatment with EB + P. These findings indicate that the attractivity that can be induced by EP + P in gonadectomized adult rats is somewhat but not wholly reduced if androgen is present during the neonatal period.     

Ovarian influences on food intake and body weight regulation in lactating rats

In the following experiments, an attempt was made to determine the role of the ovary in the control of food intake and body weight regulation during lactation. In the first study, it was found that concentrations of estradiol benzoate effective in suppressing food intake and body weight in nonlactating animals were not effective during lactation. In the second experiment, ovariectomy during lactation was shown not to produce the usual increases in food intake and body weight or change in meal patterns known to occur after ovariectomy in the nonlactating rat. These results suggested that lactating animals behave the as though functionally ovariectomized and that the removal of the ovaries is of no additional consequence. The further observation that animals nursing small litters gained weight considerably more rapidly than animals nursing large litters led to the prediction that these animals would also be more responsive to the suppressive effects of EB. In the third study, EB in concentrations which are not effective in suppressing body weight in animals nursing large litters was found to suppress body weight in mothers with small litters. However, since these animals also showed a decline in milk yield, a number of alternative interpretations of these results were considered. These results, together with data concerning levels of ovarian hormones during gestation and lactation led to the hypothesis that pregnant and lactating animals undergo an elevation in body weight set-point, similar in magnitude and quality to elevations following ovariectomy in the nonlactating animal.