Fixed-consecutive-number performance in male and female Wistar rats

The present experiment was designed to investigate whether or not response rate differences between male and female Wistar rats observed in many different experimental procedures could be attributed to sex differences in behavioral perseverance, as has been suggested by the results of previous experiments. Male and female Wistar rats were thus exposed to different fixed-consecutive-number schedules of reinforcement. Fixed-consecutive-number schedules require subjects to emit a specified number of responses on one (work) lever, before a response on another (food) lever results in the presentation of reinforcement. The response requirement on the work lever was manipulated in different experimental conditions. Subjects had to emit between 3 and 7, 8 and 12 or 13 and 17 responses on the work lever before a response on the food lever produced reinforcement. When subjects emitted fewer or more than the required number of responses on the work lever, a 5-s time-out period was presented. Males responded at higher rates than females during all experimental conditions; response rates of males and females increased as the response requirement on the work lever was increased. Sex differences in response efficiency were not observed, but males seemed to reach final response efficiency faster than females. Response efficiency decreased as the response requirement on the work lever was increased. Error analysis showed that both males and females made more errors by not producing enough responses on the work lever than by producing too many. However, males were more likely than females to emit more responses than the requirement on the work lever, while females were more likely than males not to produce enough responses on the work lever.     

Delayed spatial response alternation: effects of delay-interval duration and lesions of the medial prefrontal cortex on response accuracy of male and female Wistar rats

A delayed spatial response alternation procedure was used to assess behavioural differences between male and female Wistar rats, assumed to involve memory functioning. In Expt. I, subjects were required to alternate responses between two levers in an operant environment. The delay between response opportunities was varied between 1, 3, 7.5 and 15 s in different experimental conditions. Incorrect responses produced a time-out from experimental contingencies for the duration of the currently active delay interval. Response accuracy decreased for males as well as females as the duration of the delay interval was increased. Performance improved as subjects were exposed to the different delay interval durations during consecutive trials. Sex differences in behavioural accuracy were not observed. In Expt. II, some subjects who participated in Expt. I received lesions of the medial prefrontal cortex, while others were control-operated. When re-exposed to the 1 and 7.5 s delay conditions of the first experiment, lesioned subjects at first behaved less accurately than control-operated subjects. Accuracy, however, improved after prolonged exposure to the experimental conditions. Sex differences in behaviour after surgery could not be observed.     

Neonatal isolation enhances acquisition of cocaine self-administration and food responding in female rats

We showed previously that neonatal isolation (ISO) enhances acquisition of cocaine self-administration in adult male rats without altering acquisition of food responding. Female rats show poorer performance in learning tasks and are differentially affected by stress compared to male rats. Thus, we investigated whether ISO alters acquisition of operant responding for cocaine and food in female rats with comparison to male rats. Litters were subjected to ISO or were non-handled (NH). Activity levels were assessed in adult rats. Then, rats were implanted with jugular catheters and allowed to self-administer cocaine under a fixed-ratio 1 (FR1) schedule of reinforcement using an escalating dose presentation procedure. Cocaine intake, discrimination of active versus inactive levers, and ineffective active lever responses were tabulated. Effects of non-contingent cocaine infusions (primes) and increasing FR on responding were then assessed. Other rats were allowed to lever press for food under an FR1 schedule (10 s time-out). ISO enhanced acquisition of operant responding for food and cocaine in female rats. The latter was demonstrated by better lever discrimination, emission of fewer ineffective responses, and superior performance in response to primes. Yet, NH females ingested more cocaine than ISO females during the initial acquisition phase. In male rats, ISO enhanced acquisition of cocaine self-administration but not food responding. Activity levels were unaffected by ISO or gender. These data confirm and extend our previous findings demonstrating the enduring effects of ISO on adult self-administration behavior and emphasize the importance of measuring behavioral patterns versus intake in acquisition studies.     

Temporally spaced responding for food in adult rats after undernutrition early in life

Rats were undernourished during the suckling and early post-weaning periods and were then fed a good quality diet ad libitum from 43 to 300 days when behavioural testing was begun. Male previously undernourished and control rats were tested on three operant conditioning schedules in which low rats of bar-pressing were rewarded with food (differential reinforcement of low rate or DRL schedules).These were DRL 5 sec, DRL 10 sec and DRL 30 sec. On each schedule the response rates of both groups of animals declined over several sessions and their efficiency at obtaining the food rewards increased. However, there were no significant differences between the two groups on either of these measures.This finding indicates that, despite being hyper-responsive in many situations, including some operant conditioning schedules, previously undernourished rats are as capable as controls of witholding responsing when necessary.     

Sexual behaviour and neuronal activation in the vomeronasal pathway and hypothalamus of food-deprived male rats

As feeding and mating are mutually-exclusive goal-orientated behaviours, we investigated whether brief food deprivation would impair the display of sexual behaviour of male rats. Analysis of performance in a sexual incentive motivation test revealed that, similar to fed males, food-deprived males preferred spending time in the vicinity of receptive females rather than nonreceptive females. Despite this, food-deprived males were more likely to be slow to mate than normally-fed males, and a low dose of the satiety peptide α-melanocyte-stimulating-hormone attenuated the effect of hunger. Using Fos immunocytochemistry, we compared neuronal activity in the vomeronasal projection pathway in response to oestrous cues from receptive females between food-deprived and fed males. As in fed males, more Fos expression was seen in the rostral part of the bed nucleus of the stria terminalis and in the medial preoptic area in food-deprived males, confirming that food-deprived males can recognise and respond to female oestrous cues. However, although there was also an increase in Fos expression in the bed nucleus of the accessory tract and in the posteromedial amygdala in fed males, no increases were seen in these areas in food-deprived rats. We also found selective attenuation in the activation of lateral posterior paraventricular nucleus (lpPVN) oxytocin neurones in food-deprived males. Taken together, the data show that, although food-deprived males can still become sexually motivated, copulation is delayed, and this is accompanied by variations in neuronal activity in the vomeronasal projection pathway. We propose that, in hungry rats, the lpPVN oxytocin neurones (which project to the spinal cord and are involved in maintaining penile erection) facilitate the transition from motivation to intromission, and their lack of activation impairs intromission, and thus delays mating.     

Ghrelin's effects on food motivation in rats are not limited to palatable foods

The “hunger” hormone, ghrelin, is powerfully orexigenic. Even in the absence of hunger, ghrelin delivery to rats increases consumption of chow, as well as palatable foods, and increases motivated behaviour for palatable food rewards. Inspired by the finding that ghrelin increases the selection of chow in rats offered a choice diet (lard, sucrose or chow) and even in rats bingeing on a high‐fat diet, we aimed to explore whether the effects of ghrelin on motivation extend to regular chow. Rats were conditioned to lever press for either chow or sucrose pellets in a progressive ratio (PR) operant conditioning task. The effect of acute i.c.v. delivery of ghrelin on both chow and sucrose self‐administration was determined and compared with overnight fasting (ie, when endogenous ghrelin levels are elevated). We found that ghrelin similarly increased motivated behaviour for chow and sucrose pellets. The effect of fasting on motivated behaviour for both food pellets was comparable in magnitude to that induced by ghrelin, albeit with an earlier ceiling effect during the PR session. Devaluation experiments (in which rats are offered either food reinforcer in excess prior to PR testing) did not support the hypothesis that sucrose pellets would be more difficult to devalue (as a result of their higher incentive value) than chow pellets. When exchanging the respective pellets during a PR session, chow‐conditioned rats were more motivated for sucrose pellets compared to chow pellets; however, sucrose‐conditioned rats were similarly motivated for chow pellets compared to sucrose pellets. Thus, using sucrose as a reward may increase the motivation even for less palatable foods. We conclude that the impact of ghrelin on food‐motivated behaviour in fed rats is not limited to palatable foods but extends to regular chow, and also that the magnitude of the effect is considerable compared to that of an overnight fast.     

Undernutrition and food rehabilitation effects on the locus coeruleus in the rat

Perinatal nutrition plays a fundamental role on the morphological organization and function of a number of brain stem structures. Because little is known of the effects of perinatal undernutrition upon sexually dimorphic structures underlying reproductive behavior, the locus coeruleus morphology of 60-day-old male and female Wistar rats was analyzed. Perinatal food deprivation until weaning significantly decreased the volume and neuronal number of locus coerulus in male and female rats, while nutritional rehabilitation ameliorated these alterations in males but not in females. Data suggest that perinatal undernutrition interferes with the neuroendocrine mechanisms underlying the establishment of sex differences of the locus coeruleus.     

Genetic differences in NMDA and D1 receptor levels,and operant responding for food and morphine in Lewis and Fischer 344 rats

Previously, we have shown that Lewis (LEW) rats acquire faster than Fischer 344 (F344) rats operant food- and morphine-reinforced tasks under fixed-ratio schedules of reinforcement. The first purpose of the present work has been to study if differences in operant responding behavior may participate in the reported differences in morphine self-administration behavior between both inbred rat strains. To this end, we have analyzed the microstructure of responding obtained under a variable-interval (VI) of food reinforcement by calculating the inter-response time (IRT) for each rat strain. LEW rats exhibited shorter IRTs than F344 rats, suggesting that LEW rats may have an inherent high or compulsive operant responding activity. When subjects of both inbred rat strains were submitted to a schedule of morphine reinforcement of high responding requirements such as progressive ratio schedules, LEW rats also reached significantly higher breaking points and final response ratio than F344 rats for i.v. morphine self-administration. Given that there are neurochemical differences between both rat strains and that glutamatergic N-methyl-D-aspartate (NMDA) and dopaminergic D(1) receptors have been involved in operant responding behavior, a second purpose of this work has been to measure basal NMDA and D(1) receptor levels in these rat strains by quantitative receptor autoradiography. Compared to F344 rats, LEW rats showed higher basal NMDA receptor levels in frontal and cingulate cortex, caudate putamen, central amygdaloid nuclei, and intermediate white layer of superior colliculus, and higher basal D(1) receptor levels in several areas of hippocampus and thalamus, and substantia nigra pars reticulata. Taken together, these results suggest that an inherent high operant responding activity of LEW rats may have a role in the previous reported faster acquisition of opiate-reinforced behavior in operant self-administration paradigms under fixed-ratio schedules of reinforcement. In addition, a basal higher NMDA and D(1) receptor levels of LEW rats compared to F344 rats may participate in the neurochemical background that mediates the behavioral differences between both inbred rat strains.     

Food motivation in hypothalamic hyperphagic rats reexamined

The food motivation of rats made hyperphagic with ventromedial hypothalamic (VMH) knife cuts was examined in bar pressing tests on variable interval (VI) and fixed ratio (FR) schedules. Compared to normal rats, nondeprived VMH rats did not bar press reliably more for food during brief tests on a VI schedule, but did respond significantly more when prefed just prior to VI testing, and when tested on FR schedules. VMH rats also bar pressed more than controls during 24 hr/day tests on a FR-64 schedule. During 24 hr/day tests on a VI schedule, in which they were rewarded with entire meals, VMH rats overate by taking large meals, but they did not bar press more than controls to obtain these meals. When prevented from eating large meals the VMH rats continued to overeat by increasing their meal frequency and total VI responses, but their responses/meal again did not differ from controls. Food-deprived VMH rats did not respond more than did controls during VI or FR tests, although they ate more when food was freely available. The results indicate that under ad lib conditions VMH rats are motivated to eat larger or more frequent meals than do normal rats, although their motivation to initiate any given meal is not elevated, but that under conditions of food deprivation and reduced food accessibility VMH rats are not more motivated to eat than are normal animals.     

Sex differences in learning and inhibition in spontaneously hypertensive rats

Few studies have addressed potential differences in the nature of cognitive impairment observed in males and females with ADHD. In Experiment 1, we examined sex differences in conditioned inhibitory behaviour in spontaneously hypertensive rats (SHR strain), a purported animal model of ADHD. Rats were presented with two types of trials during each of the 15 conditioning sessions. On some trials an auditory stimulus (a tone) was presented and followed immediately by delivery of food reward. On the remaining trials the tone was preceded by presentation of a visual stimulus and on those trials food was not delivered after the tone was presented. As training progressed, conditioned responding during presentation of the tone increased on reinforced trials and decreased during the non-reinforced trials, indicative of successful discrimination and inhibition. Overall, female SHR rats exhibited less conditioned overall food cup behaviour compared to male rats. Female SHR rats also required more training sessions until they responded significantly more during presentation of the tone on reinforced trials versus non-reinforced trials. In addition, the magnitude of the discrimination was smaller in female SHR rats compared to males. In contrast, no sex differences were observed in WKY rats (commonly-used control strain) in Experiment 2. Importantly, there were no significant sex differences in baseline activity or motivation during either experiment, indicating that performance differences could not account for the observed results. These results suggest that male and female SHR rats differ in their ability to form conditioned associations and inhibit behavioural responses and may provide a useful model for sex differences in cognitive dysfunction specific to ADHD.     

Bromocriptine increased operant responding for high fat food but decreased chow intake in both obesity-prone and resistant rats

Dopamine (DA) and DA D? receptors (D2R) have been implicated in obesity and are thought to be involved in the rewarding properties of food. Osborne-Mendel (OM) rats are susceptible to diet induced obesity (DIO) while S5B/P (S5B) rats are resistant when given a high-fat diet. Here we hypothesized that the two strains would differ in high-fat food self-administration (FSA) and that the D2R agonist bromocriptine (BC) would differently affect their behavior. Ad-libitum fed OM and S5B/P rats were tested in a FSA operant chamber and were trained to lever press for high-fat food pellets under a fixed-ratio (FR1) and a progressive ratio (PR) schedule. After sixteen days of PR sessions, rats were treated with three different doses of BC (1, 10 and 20 mg/kg). No significant differences were found between the two strains in the number of active lever presses. BC treatment (10 mg/kg and 20 mg/kg) increased the number of active lever presses (10 mg/kg having the strongest effect) whereas it decreased rat chow intake in the home cage with equivalent effects in both strains. These effects were not observed on the day of BC administration but on the day following its administration. Our results suggest that these two strains have similar motivation for procuring high fat food using this paradigm. BC increased operant responding for high-fat pellets but decreased chow intake in both strains, suggesting that D2R stimulation may have enhanced the motivational drive to procure the fatty food while correspondingly decreasing the intake of regular food. These findings suggest that susceptibility to dietary obesity (prior to the onset of obesity) may not affect operant motivation for a palatable high fat food and that differential susceptibility to obesity may be related to differential sensitivity to D2R stimulation.     

Contribution of sex differences in the acute stress response to sex differences in water maze performance in the rat

Male rats outperform females in spatial tasks, such as the water maze (WM). Female rats are known to have higher basal serum corticosterone (CORT) levels and to manifest a more rapid and stronger CORT response to novel stressors. Sex differences in stress responses to the handling and forced swimming in the WM task might contribute to the sex difference in WM performance. In Experiment 1, naive females were found to be impaired relative to naive males in swimming to a visible platform in a WM pool due to strongly thigmotaxic swimming by females. In Experiment 2, serum CORT, a physiological measure of stress, was highly elevated during and after WM training, with female > male values and strong inverse correlations between CORT and measures of WM performance in females. Familiarization with the WM pool and test procedures by strategies pretraining prior to spatial training reduced or eliminated the sex differences in the stress response and WM performance. In Experiment 3, adrenalectomy to eliminate the stress response eliminated sex differences in WM performance. Taken together, the results suggest that male and female rats may harbor brain circuitry that is equally capable of accurate spatial navigation and memory in the WM but which may be impaired to different degrees by the differential stress responses triggered by WM testing.     

Effects of the 5-HT1A agonist 8-OH-DPAT on food intake in food-deprived rats.

The effects of 8-OH-DPAT were investigated on food intake in food-deprived rats in both non-operant and operant feeding paradigms. 8-OH-DPAT produced dose-related reductions in food intake in both paradigms. While the higher doses of 8-OH-DPAT used in this study (i.e. 125-500 micrograms kg-1) produced a number of stereotyped behaviours which may have interfered with normal feeding, these behaviours were not apparent with the lower doses (i.e. 15.625-62.5 micrograms kg-1) which also depressed food-intake. The results of this study thus suggest that the inhibitory effect of 8-OH-DPAT on feeding in food-deprived rats is not secondary to the induction of stereotypy.     

Male and female Wistar rats differ in decision-making performance in a rodent version of the Iowa Gambling Task

The Iowa Gambling Task (IGT) measures decision-making processes by simulating real-life decisions involving reward, punishment, and uncertainty of outcomes. In humans, men show more choices for the advantageous option than women. Here, we investigated sex differences in a rat model of the IGT (r-IGT). In our r-IGT mildly food-deprived rats learn to differentiate a long-term advantageous arm from a long-term disadvantageous arm differing in frequency and amount of sugar pellets as well as unpalatable but not uneatable quinine-treated sugar pellets. We also used a T-maze discrimination procedure in which rats learn to differentiate a high from a low reward arm to further explore sex differences in reward-related decision-making. In line with human data, male rats showed a stronger task progression of choices for the advantageous option than female rats. Furthermore, male rats showed more win-stay and less lose-shift behaviour in the advantageous arm as the task progressed than female rats. Whilst both male and female rats had a stronger preference for the high over the small reward arm in the T-maze, males increased this preference over sessions, whilst females did not. These data are discussed in relation to sex differences in processing rewards and punishments.     

Central injections of growth hormone-releasing factor increase operant responding for food reward

1. Male Wistar rats were prepared with chronic ventricular cannulae and trained to bar press for food pellets in operant cages. 2. Some rats were given icv injections of 0 (vehicle), 0.4, 4.0 and 40.0 picomole GRF on separate days and tested on a continuous reinforcement (CRF) reward schedule. 3. Other rats were given icv injections of 0 (vehicle) and 4 picomole GRF on separate days and tested on a progressive ratio (PR) reward schedule. 4. In comparison with vehicle injections, GRF significantly increased the average response rate under the CRF schedule and the average breaking point under the PR schedule. 5. The increased operant responsiveness displayed by rats suggests that central GRF possesses motivational efficacy, increasing the relative reinforcing strength of food.     

Age-associated sex differences in response to food deprivation in two animal tests of anxiety

The effects of mild food deprivation (7 days of food restricted to once daily feeding to maintain body weights at 85% of free-feeding weights) were examined in adult male and female and adolescent female rats tested in the elevated plus maze and social interaction tests of anxiety. In adult male rats, food deprivation appeared to have an anxiolytic effect in the plus-maze as it significantly increased the percentage of entries onto open arms and the percentage of time spent on the open arms, without changing the number of closed arm entries. There were no effects of food deprivation in adult females, although in adolescent females food deprivation significantly increased the percentage of open arm entries rats. Adolescent female rats have female brains, but do not have circulating gonadal hormones and thus these results suggest that circulating female gonadal hormones are able to suppress some of the effects of mild food deprivation in the plus-maze. In the social interaction test, there were no effects of food deprivation in any group on the time spent in social interaction. There were opposite effects on locomotor activity in the adult male and female rats, with deprivation increasing activity in males and decreasing it in females. There were no effects of food deprivation on locomotor activity in the adolescent females, suggesting that circulating gonadal hormones were responsible for the bidirectional effects in the adult rats. In both tests there were age-associated differences in the female rats, with the adolescent females being less anxious (higher percentage of open arm entries and increased social interaction) than the adults.     

Neonatal handling affects learning, reversal learning and antioxidant enzymes activities in a sex-specific manner in rats

Early life experiences have profound influences on behavior and neurochemical parameters in adult life. The aim of this study is to verify neonatal handling-induced sex specific differences on learning and reversal learning as well as oxidative stress parameters in the prefrontal cortex and striatum of adult rats. Litters of rats were non-handled or handled (10 min/day, days 1-10 after birth). In adulthood, learning and reversal learning were evaluated using a Y maze associated with palatable food in male and female rats. Morris water maze reversal learning was verified in males. Oxidative stress parameters were evaluated in both genders. Male neonatal handled animals had a worse performance in the Y maze reversal learning compared to non-handled ones and no difference was observed in the water maze reversal learning task. Regarding females, neonatal handled rats had a better performance during the Y maze learning phase compared to non-handled ones. In addition, neonatal handled female animals showed a decreased SOD/CAT ratio in the PFC compared to non-handled females. We conclude that neonatal handling effects on learning and memory in adult rats are sex and task specific. The sex specific differences are also observed in the evaluation of antioxidant enzymes activities with neonatal handling affecting only females.     

The development of sex differences in the locus coeruleus of the rat

Development of sex differences in the locus coeruleus (LC) is investigated. The LC is a sexually dimorphic structure in which the female manifests a larger volume and greater number of neurons than do males. Male and female Wistar rats were sacrificed on prenatal days (E) 16 and 20 and postnatally (P) on days 1, 3, 7, 15, 35, 45, 60, and 90. Male and female rats show a continuous increase in the number of neurons after birth that stops in the males by P45 and in females by P60. These findings point out the existence of different patterns of development in male and female rats and may suggest that sex differences could be established because of the existence of a differential period of neurogenesis in both sexes in the postpubertal period.     

Genetic differences in NMDA and D1 receptor levels, and operant responding for food and morphine in Lewis and Fischer 344 rats

Previously, we have shown that Lewis (LEW) rats acquire faster than Fischer 344 (F344) rats operant food- and morphine-reinforced tasks under fixed-ratio schedules of reinforcement. The first purpose of the present work has been to study if differences in operant responding behavior may participate in the reported differences in morphine self-administration behavior between both inbred rat strains. To this end, we have analyzed the microstructure of responding obtained under a variable-interval (VI) of food reinforcement by calculating the inter-response time (IRT) for each rat strain. LEW rats exhibited shorter IRTs than F344 rats, suggesting that LEW rats may have an inherent high or compulsive operant responding activity. When subjects of both inbred rat strains were submitted to a schedule of morphine reinforcement of high responding requirements such as progressive ratio schedules, LEW rats also reached significantly higher breaking points and final response ratio than F344 rats for i.v. morphine self-administration. Given that there are neurochemical differences between both rat strains and that glutamatergic N-methyl- -aspartate (NMDA) and dopaminergic D₁ receptors have been involved in operant responding behavior, a second purpose of this work has been to measure basal NMDA and D₁ receptor levels in these rat strains by quantitative receptor autoradiography. Compared to F344 rats, LEW rats showed higher basal NMDA receptor levels in frontal and cingulate cortex, caudate putamen, central amygdaloid nuclei, and intermediate white layer of superior colliculus, and higher basal D₁ receptor levels in several areas of hippocampus and thalamus, and substantia nigra pars reticulata. Taken together, these results suggest that an inherent high operant responding activity of LEW rats may have a role in the previous reported faster acquisition of opiate-reinforced behavior in operant self-administration paradigms under fixed-ratio schedules of reinforcement. In addition, a basal higher NMDA and D₁ receptor levels of LEW rats compared to F344 rats may participate in the neurochemical background that mediates the behavioral differences between both inbred rat strains.     

Sex differences in skilled movement in response to restraint stress and recovery from stress

Sex differences exist in skilled movement, and skilled motor performance is also influenced by stress. As shown for cognitive function, the effects of stress are usually characterized by considerable sexual dimorphism. The purpose of this study was to investigate sex differences in skilled motor function in response to stress. Male and female Long-Evans rats were trained and tested in skilled reaching and skilled walking tasks. Both groups of animals were then exposed to daily restraint stress for 15 days. Recovery from daily stress was assessed by comparing reaching performance at 10 min versus 60 min after restraint stress, and recovery from chronic stress was tested for 21 days after cessation of stress. Animals were tested daily in skilled reaching for the entire period. Observations showed that females performed significantly better than males during the stress period in terms of reaching success and number of attempts needed to grasp a food pellet. No difference between testing at 10 or 60 min after daily stress was found. Analysis of movement patterns and recovery from stress indicated that males and females use different strategies to overcome stress-induced motor disturbance. While male rats preferred to use original movement patterns, females tended to modify these patterns in order to increase reaching success. Modification of movement patterns in female rats was accompanied by a faster recovery in success rate after the cessation of stress. These results indicate sex differences in skilled reaching in response to stress, and in the recovery period after stress.