Spatial learning deficits induced by muscimol and CL218,872: lack of effect of prenatal malnutrition.

The sensitivity of prenatal protein malnourished rats to the amnestic properties of the direct GABAA receptor agonist muscimol and the selective benzodiazepine (BZ) receptor agonist, CL218,872, was studied in the male offspring of rats provided with a protein deficient diet (6% casein) for 5 weeks prior to mating and throughout pregnancy. At postnatal day 90, rats were tested during acquisition of the submerged platform version of the Morris water maze task using four systemic doses of muscimol (0.1, 0.3, 1.0 and 1.8 mg/kg i.p.) or three systemic doses of CL218,872 (1.0, 3.2, and 5.6 mg/kg i.p.). In a dose dependent manner both drugs impaired acquisition of the task and impaired accuracy of the search pattern on the probe trial (platform removed). However, neither drug dissociated the performance of the two nutritional groups. These data are important in light of previous findings of differential behavioral effects of the non-specific BZ agonist, chlordiazepoxide (CDP), on spatial learning and on drug discrimination in prenatally malnourished rats and in the context of previous findings of reduced sensitivity to the anxiolytic effects of non-specific BZ receptor agonists across a wide variety of models of malnutrition. The present findings also support the concept that prenatal malnutrition does not affect the global functioning of the GABAA receptor, but fundamentally alters the way in which a subset of GABAA receptors (i.e. those containing the alpha2, alpha3 and/or the alpha5 but not the alpha1 subunit) is modulated by BZs.

Chlordiazepoxide-induced spatial learning deficits: dose-dependent differences following prenatal malnutrition.

The sensitivity of prenatally protein-malnourished rats to the amnestic properties of the benzodiazepine (BZ) receptor agonist, chlordiazepoxide (CDP), was studied in the male offspring of rats provided with a protein-deficient diet (6% casein) for 5 weeks prior to mating and throughout pregnancy. Rats were tested during acquisition of the submerged platform version of the Morris water maze task using three systemic doses of CDP (3.2, 5.6, and 7.5 mg/kg i.p.) at two ages (day 30 and day 90). At 30 days, prenatally malnourished rats showed less sensitivity to the amnestic effect of the 5.6-mg/kg dose when compared with well-nourished controls by displaying shorter swim paths during acquisition and a more selective search of the target quadrant upon removal of the platform (probe trial). At 90 days, prenatally malnourished rats again showed less sensitivity to CDP at a dose of 5.6 mg/kg, but more sensitivity to the 3.2-mg/kg dose (indicated on the probe trial). No obvious relationship was identified between the nutritional group differences in behavioral sensitivity to CDP at 90 days and their BZ receptor density in the hippocampus or medial septum. It can be concluded that prenatal malnutrition alters the amnestic response to CDP in a dose-dependent and developmentally specific manner, thus providing further support for functional changes within the GABAergic system subsequent to malnutrition.

Prenatal protein restriction increases sensitization to cocaine-induced stereotypy.

The present study characterized the total amount of stereotyped behavior following acute and repeated administration of cocaine in male and female prenatally protein malnourished rats. Adult offspring of female Sprague-Dawley rats fed either a low (6% casein) or adequate (25% casein) protein diet 5 weeks prior to mating and throughout their pregnancy were studied. Once every 3 days (for a total of six injections), half the rats from each nutritional treatment group (repeated exposure) were injected with cocaine (30 mg/kg, i.p.) and their total amount of stereotypy (rearing, forepaw treading, compulsive sniffing and head bobbing) monitored. The remaining rats received five saline injections followed by a cocaine injection on the last injection day (acute exposure group) and their behavioral response was also measured. Despite being slightly less sensitive to cocaine following their first injection, by the sixth injection, prenatally protein malnourished animals in the repeated-exposure group exhibited significantly greater sensitization to the psychomotor stimulant effects of cocaine than well-nourished controls. In the acute exposure groups, however, prenatally malnourished males, but not females, exhibited significantly more stereotypy than well-nourished subjects following a single cocaine injection. These findings have implications for characterizing addiction potential in the previously malnourished rats, as well as providing additional information regarding factors which can influence sensitization.

Differential Effects of Prenatal Protein Malnutrition and Prenatal Cocaine on Radial Arm Maze Performance in Adult Male Rats.

The effects of prenatal cocaine and protein malnutrition were examined on acquisition of the radial arm maze in adult male Sprague-Dawley rats whose mothers were provided with a 6% casein, a 25% casein or a standard chow diet and cocaine (30mg/kg) or saline injections beginning 5 weeks prior to mating and continuing to parturition. Rats were tested using an 8-arm radial maze with 4 baited arms and were required to collect all 4 food pellets within 5 min to complete a trial. Subjects were tested for 1 trial/day until they met criterion for successful acquisition of the task. Criterion was attained when the rat collected 3 out of the 4 food pellets within their first 4 arm entries within a trial (while still completing the trial) with this level of performance being maintained for 3 consecutive trials. The results showed dissociation between the effects of prenatal protein malnutrition and prenatal cocaine. Prenatally protein malnourished adult male rats required a greater number of trials to criterion, made more reference memory (but not working memory) errors, and required a longer time to complete each trial when compared with control males. However, rats with prenatal cocaine exposure showed no significant impairments in the radial arm maze. These results contrast with our previous findings using the Morris maze task in which adult male subjects exhibited impaired acquisition following prenatal cocaine while there were no effects following prenatal protein malnutrition. Thus, the radial arm maze and water maze procedures appear to engage different processes that are differentially sensitive to the prenatal insults.

Effects of diet on sensitization to cocaine-induced stereotypy in female rats.

The progressive increase in cocaine-induced stereotyped behavior that accompanies repeated cocaine injections (sensitization) was examined in rats consuming different diets. Adult female Sprague-Dawley rats were fed one of three diets: low protein (6% casein), adequate protein (25% casein), or a standard chow diet. Following 1 week of adaptation to the diets, the rats were injected every 3-4 days with either cocaine (30 mg/kg, IP) or saline, and the total amount of stereotypy was measured over a 90-min interval following each of four injections. Cocaine-induced stereotypy peaked at 40-50 min following each injection, after which it declined for all diet groups. With repeated injections, the total amount of stereotypy increased in all diet groups. By the fourth injection, the low protein diet group (6% casein) exhibited a slower onset and a possibly prolonged duration of cocaine-induced stereotypy when compared with the two adequate protein diet groups (25% casein and chow). Interestingly, the rats in the two purified diet groups (6% casein and 25% casein) exhibited significantly more stereotypy across injections than those in the chow diet group. Weight differences did not explain the differences in stereotypy present among the diet groups. This study concludes that diet significantly alters the pattern of cocaine-induced stereotypy in female rats, especially after repeated exposure.

Development of spatial navigation following prenatal cocaine and malnutrition in rats: lack of additive effects.

The effects of prenatal cocaine exposure and protein malnutrition on the development of spatial navigation were assessed in rats. Sprague-Dawley dams were fed a low-protein (6% casein), adequate protein (25% casein), or a laboratory chow diet prior to mating and throughout pregnancy. Within each diet group, rats received either cocaine injections (30 mg/kg i.p. two times per week prior to mating and then 30 mg/kg s.c. daily from day 3 to 18 of pregnancy) or saline injections. All litters were fostered on the day of birth to saline-injected mothers fed either the 25% casein diet or the chow diet. Gestation length was decreased by prenatal cocaine exposure whereas litter size was reduced by prenatal malnutrition. On postnatal days 21, 25, 30, or 70, rats were tested for their ability to locate a submerged platform in a Morris water maze. In well-nourished rats, prenatal cocaine increased the mean distance swum during acquisition over days 21-30, a difference that was abolished in rats with prenatal malnutrition. In the absence of drug exposure (saline groups), prenatal malnutrition was itself associated with longer swim paths. Neither prenatal insult affected the accuracy of the spatial navigation at these ages, as determined by their search pattern when the platform was removed. On postnatal day 25, rats raised on the chow diet exhibited superior performance to that of rats raised on the 25% casein diet, but by day 30 these two well-nourished groups were comparable. At day 70, prenatal cocaine impaired spatial performance on the first session, in well-nourished rats only. Thus, these results provide no support for the hypothesis that prenatal cocaine and protein malnutrition combine to produce a greater effect on behavioral development than either insult alone.

Presession noise increases sensitivity to chlordiazepoxide's discriminative stimulus in pigeons.

1. Pigeons were trained to discriminate chlordiazepoxide (CDP) from saline using two-key food reinforced drug discrimination procedures. Discriminative control by CDP was maintained despite extended training with vehicle-like doses of CDP, by using a modified "fading" procedure that provided for a mixture of drug discrimination training sessions preceded by an i.m. injection of either 8.0 mg/kg CDP, or a lower training dose of CDP (4.0, 2.8, 2.0, 1.4, 1.0, 0.7, or 0.5 mg/kg CDP), or saline. The lower training dose was decreased across blocks of sessions. 2. Four lower training doses (1.4, 1.0, 0.7, and 0.5 mg/kg CDP) were retrained, with 10 min of 98 dB of noise administered 75 min prior to each drug discrimination training session. Presession exposure to noise increased percent CDP-appropriate choices for each of the four lower training doses by 15-20% over those obtained previously. 3. It is concluded that brief presession exposure to loud noise increases sensitivity to the discriminative stimulus effects of low training doses of CDP.

Prenatal cocaine but not prenatal malnutrition affects foster mother-pup interactions in rats.

The separate and combined effects of prenatal cocaine exposure and malnutrition on mother-pup interactions in rats were assessed daily from postnatal day 2 to day 21. Sprague-Dawley dams were fed a diet of low protein content (6% casein), an isocaloric diet of adequate protein content (25% casein, control), or a laboratory chow diet prior to mating and throughout pregnancy. Within each diet group, rats received either cocaine injections (30 mg/kg IP two times per week prior to mating and then 30 mg/kg SC daily from days 3 to 18 of pregnancy) or saline injections. Litters were fostered on the day of birth to control mothers (i.e., nondrug-exposed dams fed the control or chow diet). Foster mothers fed the 25% casein diet showed increased contact with cocaine-exposed pups compared with nondrug-exposed pups in the second postnatal week but lower levels as the pups approached weaning. Passive nursing was increased in dams caring for prenatally malnourished, cocaine-exposed pups compared with those caring for similar pups with no drug exposure. Chow-fed mothers did not differ in their behavior towards pups with or without prenatal cocaine treatment. Prenatal cocaine and malnutrition independently compromised birth weight and various reflexive milestones but the attainment of physical milestones was affected only by prenatal cocaine. There were no additive effects of the two prenatal insults on any measure of mother-pup interaction or pup development.

The effects of cocaine exposure prior to and during pregnancy in rats fed low or adequate protein diets.

Sprague-Dawley rats were fed a diet of low protein content (6% casein), an isocaloric diet of adequate protein content (25% casein), or a laboratory chow diet for 5 weeks prior to mating and throughout pregnancy. Within each diet group, rats received either cocaine (30 mg/kg IP two times per week prior to mating and 30 mg/kg or 40 mg/kg SC daily from days 3 to 18 of pregnancy) or saline injections. Cocaine produced a greater reduction in food intake during pregnancy in the malnourished group compared with the other two diet groups. The effect of cocaine on food intake was minimal in chow-fed rats. Weight gain in pregnancy was reduced by cocaine in a dose-dependent manner, and by malnutrition. Both prenatal cocaine and malnutrition impaired skeletal maturation of the pups, but there was no additive effect of the two insults on this measure. Litter size was significantly reduced by the 40 mg/kg, but not by the 30 mg/kg dose of cocaine across all diet groups. Consequently, the 40 mg/kg dose of cocaine proved to be fetotoxic in this model. Birth weight was significantly reduced by prenatal malnutrition but not by prenatal cocaine. Gestation length was unaffected by either insult. Hence, the ability to detect a diet x drug interaction was dependent upon the variable being measured.

Effects of an every other day rapid kindling procedure in prenatally protein malnourished rats.

Prenatally protein (6/25) rats have been reported to require significantly more stimulations to attain a stage 5 seizure than well-nourished controls (25/25) when using either a traditional or rapid every day, kindling procedure. In the present study, a rapid kindling procedure was utilized where both prenatally malnourished and control rats received every other day perforant path kindling (50 Hz, 10 s train) 12 times a day at 5-min intervals. Using this procedure, stage 5 seizures and a fully state were attained in both nutritional groups at approximately the same rate. It is postulated that it is the every other day component of the present procedure which overcomes seizure-induced inhibition in the 6/25 subjects, thereby allowing them to attain stage 5 seizures at the same rate as controls.

Drug discrimination training with low doses: maintenance of discriminative control.

Procedures are reported that maintain control by the drug cue during and after drug discrimination training with lower doses that yield predominantly vehicle-appropriate choices. Twelve pigeons were trained to discriminate chlordiazepoxide (CDP) from saline using two-key (drug vs. vehicle) drug discrimination procedures. Intermixed within each block of 30 sessions were nine sessions of training with 8.0 mg/kg CDP, nine with one of seven lower training doses (4.0, 2.8, 2.0, 1.4, 1.0, 0.7, or 0.5 mg/kg CDP), and 12 with saline. The lower training dose was decreased across blocks. The three lowest training doses (1.0, 0.7, and 0.5 mg/kg CDP) yielded predominantly saline-appropriate choices but had no effect on discrimination of 8.0 mg/kg CDP or saline. Three doses (2.0, 1.4, and 1.0 mg/kg CDP) were retrained, and each yielded percentages of drug-appropriate choices nearly identical to those obtained during previous training. This drug discrimination procedure maintains control by the drug cue during and after training with vehicle-like doses of the training drug and may allow for repeated assessment of effects of low training doses.