Cholecystokinin conditioning in rats: ontogenetic determinants

Low doses (0.12-2.0 micrograms/kg) of cholecystokinin octapeptide (CCK), administered intraperitoneally, support the formation of conditioned odor preference in neonatal and weanling rats. Exposure to a novel odor was paired with CCK injection, and the rats' olfactory choices were assessed 24 hr later. Rats at 5, 11, and 22 days of age preferred the odor previously associated with CCK, compared with vehicle-injected littermates. In contrast, CCK failed to support olfactory conditioning in 28-day-old rats, whether they were (a) weaned and independently housed, (b) residing with the dam and suckling, or (c) fed only milk. Adult rats also did not establish an odor preference with CCK as the unconditioned stimulus. Thus, CCK's changing impact from positive to neutral probably occurs during the rats' 4th postnatal week and may be related to maturational changes occurring during the final stages of weaning.     

Defensive burying following injections of cholecystokinin, bombesin, and LiCl in rats.

Past research indicates that feeding is reduced for animals injected with cholecystokinin and bombesin. One explanation for this effect suggests that these peptides act as natural satiety signals; an opposing view asserts that bombesin and cholecystokinin reduce feeding through malaise. The present experiment tested the basic assumptions associated with these positions using the defensive burying procedure. Groups of rats were given sweetened condensed milk followed by IP injections of bombesin (6, 16, and 32 micrograms/kg), cholecystokinin (0.7, 1.4, and 2.9 micrograms/kg), LiCl (6.4 mg/ml), or saline. The results showed that animals injected with cholecystokinin, bombesin, and LiCl developed learned aversions to the milk and actively buried the milk spout with their bedding. The findings provide further support for the view that bombesin and cholecystokinin induce malaise rather than satiety.     

An ontogenetic comparison of ethanol-mediated taste aversion learning and ethanol-induced hypothermia in preweanling rats.

To examine the interactive effects of ethanol (EtOH) and ambient temperature, 10-, 16-, and 20-day-old rat pups ingested pairings of sucrose solution and various doses of ethanol (intubated intragastrically) and were then exposed to relatively low or relatively high ambient temperatures. Ten- and 20-day-old pups required a higher EtOH dose than did 16-day old pups for conditioning of a sucrose aversion and for hypothermia. These age-related differences might be due to ontogenetic changes in the production and accumulation of acetaldehyde, a metabolite of EtOH. For all ages, EtOH-induced hypothermia was necessary for conditioning of the taste aversion, which is in accord with results of previous tests with adult rats (Cunningham, Hawks, & Niehus, 1988).     

Regeneration in the optic nerve of adult rats: influences of cultured astrocytes and optic nerve grafts of different ontogenetic stages

Summary We have studied the effects of transplanted optic nerves of different ontogenetic stages (E19 to adult), and cultured astrocytes from P2 cerebral cortex on the regeneration of axons in the optic nerve of adult rats. Regeneration was visualized by anterograde tracing with rhodamine-iso-thiocyanate. Grafts were identified with Nuclear Yellow. Astroglia within both the cut optic nerve and the transplants were detected by anti-glial fibrillary acidic protein staining. In control animals (cut optic nerve, 2–3 mm behind the optic disc), only a few neurites were found 15 days after the operation which grew randomly for short distances into the surrounding meningeal sheaths. Perinatal (E19 to P2) optic nerves induced a massive outgrowth of RITC-filled axons from the host optic nerve. The regenerating fibres grew for up to 3 mm towards the graft, ahead of glial fibrillary acidic protein-positive astroglia emanating from the host optic nerve that seemed to follow them. Although the regenerating fibres reached the grafts, they did not penetrate them. Optic nerve grafts of increasing age elicited smaller growth responses; e.g. grafts from P8 promoted only a very limited (several 100 m) growth response, grafts from P12 and later induced outgrowth comparable with that of control animals. Grafted astrocytes from P2 donors that had previously been grown in culture, were also capable of promoting outgrowth of rhodamine-iso-thiocyanate-filled axons from the host optic nerve. These findings suggest that only astrocytes at an immature stage of differentiation are capable of inducing axon growth from the adult optic nerve. Furthermore, the absence of an obvious cellular bridge between host and graft suggests that the graft effect is probably mediated by the release of astroglia-derived diffusible neurite growth promoting factors.     

Nod2 influences microbial resilience and susceptibility to colitis following antibiotic exposure

Inflammatory bowel disease (IBD) etiology involves genetic susceptibility, environmental triggers, and the gut microbiome. Antibiotic exposure is associated with IBD, both in early life and adulthood. Here, we investigated whether Nod2-deficiency influenced response of the gut microbiota to antibiotics and subsequent colitis susceptibility. Wild-type and Nod2^−/− littermate mice were treated with amoxicillin as adults or neonates, and fecal samples were collected for 16S rRNA sequencing. Five weeks after antibiotic exposure, dextran sulfate sodium (DSS) colitis was induced. Antibiotic treatment altered the microbiota of adult WT and Nod2^−/− mice, but recovery was delayed in Nod2^−/− mice. Neonatal antibiotic treatment significantly changed the microbiota at weaning in WT and Nod2^−/− littermates; however, Nod2^−/− mice maintained reduced microbial diversity 14 days after cessation of antibiotics. Although treatment of adult mice did not influence susceptibility to colitis, neonatally treated Nod2^−/− mice developed a more severe colitis. Moreover, the colitis phenotype was transferable through fecal transplantation into germ-free Nod2^−/− recipients, and was associated with changes in intestinal T cells and the cytokine milieu following inflammation. These data demonstrate that neonatal antibiotic exposure has long-lasting influence on the microbiota and mucosal immunity, and may explain how NOD2 contributes to the risk of intestinal inflammation.     

Environmental influences on locomotor recovery following cortical lesions in rats

Groups of rats were exposed to an enriched environment 2 hr per day for 30 days during the immediate pre- and/or postoperative period, or not at all. Animals in four of these groups sustained lesions in the bilateral sensorimotor cortex. One sham-operated control group was enriched pre- and postoperatively; a second control group was not. Animals were trained preoperatively to locomote across a narrow elevated runway. Testing on the locomotor task began 31 days after surgery and continued until preoperative performance levels were achieved. Preoperative enrichment had the most potent influence on initial deficit and recovery of locomotion. Animals that were enriched preoperatively failed to demonstrate any deficit postoperatively, and the topology of their hindlimb movement appeared to be normal. In preoperatively impoverished animals, postoperative enrichment reduced the degree of initial deficit and speeded recovery of locomotion when compared with animals not enriched at all. However, preoperatively impoverished rats demonstrated an aberrant topology of hindlimb movement even after they were "behaviorally recovered".     

Acetaminophen-induced forestomach lesion in normal rats following intravenous exposure

Four groups of ten male and ten female rats each were treated intravenously with saline, 400 mg/kg/day of a commercially available injectable acetaminophen formulation, or 400 mg/kg/day of a new injectable acetaminophen formulation with (aged) or without (fresh) impurities daily for fourteen days. Gross observations of the mucosal surface of the stomachs from treated rats included multifocal to diffuse pale, elevated foci confined to the nonglandular region of the stomach. Treatment-related histologic observations consisted of epithelial hyperplasia and hyperkeratosis of the nonglandular mucosa of the stomach. The epithelial hyperplasia was characterized by a thickened epithelium, frequently accompanied by the development of undulations at the basement membrane zone, resulting in the formation of rete ridgelike structures protruding into the underlying tissue. The submucosa was usually expanded by edema and occasionally contained an infiltrate of neutrophils, eosinophils, and macrophages. The hyperplasia was usually accompanied by hyperkeratosis resulting in thickening of the stratum corneum. The incidence and severity of the gastric changes were similar across all treatment regimens. Although considered clinically irrelevant since humans do not have a forestomach equivalent, these results are significant in that this appears to be the first report of forestomach hyperplasia and hyperkeratosis following intravenous exposure to acetaminophen.     

Maternal and sex-related influences on locomotor activity in rats following weaning

The effect of the presence of the mother beyond the normal time of weaning on the locomotor activity of litters is evaluated in the present work. Weaned rats, either females or males, showed a significantly higher activity in the open-field arena than the non-weaned ones. A sex-related difference was also detected. Weaned and nonweaned females showed higher activity than the weaned and nonweaned males, respectively. The androgen-induced lowering of female activity is affected by the presence of the mother. This is substained by the fact that weaned androgenized females exhibited higher levels of activity than nonweaned androgenized females. Our observations suggest that the presence of the mother after the normal time of weaning disturbs the emotional maturation of litters.     

State-dependent learning following electrical stimulation of the hippocampus: intact and split-brain rats

In experiment 1, electrical stimulation of the posterior hippocampus was shown to produce state-dependent learning (SDL) for a step-out inhibitory avoidance task in rats. Stimulation sites in either the right or left hippocampus were equally effective in producing this effect. Similarly, the presence or absence of afterdischarge (AD) following the stimulation did not differentially affect performance on the task. In experiment 2, forebrain bisection ameliorated the behavioral deficits in the animals receiving stimulation before testing but not before training (N/S group), while those stimulated before training but not before testing (S/N group) remained impaired; thus, providing a demonstration of asymmetrical SDL. Variations in extent of the commissurotomy differentially affected the laterality of the afterdischarge but not the performance in the SDL task. Speculation as to the mechanisms of this SDL effect was presented.     

Adolescent exposure to nicotine impairs adult serial pattern learning in rats

In the present study investigating the effects of adolescent nicotine exposure on adult serial pattern learning, adolescent rats received daily i.p. injections of either 1.0 mg/kg nicotine or saline for 5 days per week for 5 weeks beginning on postnatal day 25 (P25), then were allowed 35 days drug free. Rats then began training on P95 as adults on a 24-element serial pattern composed of eight 3-element chunks. Adolescent exposure to 1.0 mg/kg nicotine produced persistent retardation of learning for the first element of each 3-element chunk of the pattern, that is, for chunk boundary elements, and transient retardation of learning for elements 2 and 3 of each chunk of the pattern, that is, for the within-chunk elements. Deficits at chunk boundaries were interpreted as deficits of phrasing cue discrimination learning whereas deficits for learning responses for elements within-chunks (elements 2 and 3 of chunks) were interpreted as deficits of rule learning. These results indicate that the effects of adolescent nicotine exposure on adult learning and cognitive capacity deserve further scrutiny.     

Choline supplementation following third-trimester-equivalent alcohol exposure attenuates behavioral alterations in rats

Despite the known adverse consequences of prenatal alcohol exposure, some pregnant women continue to drink alcohol, making it imperative to identify treatments for children with fetal alcohol spectrum disorders. The authors recently reported that perinatal choline supplementation can reduce some fetal alcohol effects (J. D. Thomas, M. Garrison, & T. M. O'Neill, 2004), and the present study examined whether choline supplementation is effective when administered after third-trimester-equivalent ethanol treatment. Rat pups were exposed to 6.0 g/kg/day ethanol during the neonatal brain growth spurt (Postnatal Days [PD] 4-9) and treated with choline chloride (0, 10, 50, or 100 mg/kg) from PD 10-30. Behavioral testing occurred after choline treatment had ceased. Female subjects exposed to ethanol were overactive and exhibited spatial learning deficits, effects that were attenuated with all doses of choline supplementation. These data indicate that choline supplementation can alter brain development following a developmental insult. Moreover, the data suggest that early dietary interventions may reduce the severity of some fetal alcohol effects, even when administered after birth.     

Altered sensitivity to NMDA following developmental lead exposure in rats.

Early Pb exposure is known to disrupt the development of the hippocampus and result in deficits in learning and memory capacities and altered seizure susceptibility. The excitatory amino acid, NMDA, is found in high concentrations in the hippocampus and has been implicated in learning and memory functions and seizure activity. Rat pups nursed mothers exposed to high (4%), moderate (0.4%), or low (0.05%) levels of PbCO3 in their diet, or a Na2CO3 control diet from postnatal day 1 (P1) to P25. Rat pups were injected with varying doses of NMDA on P15 or P25. Control animals showed a characteristic slowly developing response to NMDA, usually including tail twitches and wet dog shakes at approximately 10 and 40 mg/kg at P15 and P25, respectively, with status epilepticus and death occurring at 40 and 80 mg/kg. Lead-exposed animals displayed an altered sensitivity to NMDA, with high and medium Pb animals showing the onset of behavioral signs and death at lower NMDA doses, the degree of which being dependent on the level of Pb exposure. Low Pb-exposed animals showed a more variable and attenuated response to NMDA. The data are discussed in terms of the possible mechanisms of Pb neurotoxicity.     

Flavor avoidance induced by LiCl and dexfenfluramine in rats and mice using nondeprivation protocols

Using dexfenfluramine as unconditional stimulus (US), the authors confirmed that sham-operated and area postrema (AP)-lesioned rats form comparable conditioned flavor avoidances. When lithium chloride (LiCI) was used as the US, AP-lesioned rats did not learn to avoid a drug-paired flavor conditional stimulus (CS+). Sham-operated, but not AP-lesioned, rats had low intakes of the placebo-paired flavor (CS-), which suggests that the lesions disrupted generalization of avoidance. Generalized avoidance in intact rats was similar when either sweetened milk or Polycose was used as the caloric vehicle for the CSs. When flavored gels of Polycose were used as CSs, C57BL/6J mice developed flavor avoidance with either LiCl or dexfenfluramine as US. Compared with rats, mice required higher doses of these agents, avoidance was not complete after many pairings, and there was no generalization to the CS-.     

Critical periods for the effects of alcohol exposure on learning in rats

Critical periods for alcohol-induced deficits in spatial navigation and passive avoidance learning were investigated with a rat model of fetal alcohol syndrome. Rats were exposed to alcohol prenatally (Gestational Days 1-10 or 11-22) or postnatally (Postnatal Days 2-10) or throughout all 3 periods. Offspring were tested in either a spatial navigation or an avoidance task as juveniles or adults. As juveniles, the combined exposure group took longer to learn the spatial navigation task compared with all other groups. This effect was not seen in adults. Passive avoidance performance was not affected. These results suggest that long-term exposure to alcohol during development has adverse effects on spatial learning. The lack of differences in the short-term exposure groups implies that there may not be 1 critical period of alcohol exposure, but that the adverse effects of alcohol during development may be cumulative on some behaviors.     

Cholinesterase inhibitors ameliorate spatial learning deficits in rats following hypobaric hypoxia

Cognitive functions especially learning and memory are severely affected by high altitude (HA) exposure. Hypobaric hypoxia (HBH) encountered at HA is known to cause oxidative stress, alterations of neurotransmitters and cognitive impairment. We hypothesized that alteration in cholinergic system may be involved in HBH-induced learning impairment. The present study has investigated the cholinergic dysfunctions associated with simulated HBH-induced impairment of learning in rats and protective role of acetylcholine esterase inhibitors (AChEIs). Male Sprague–Dawley rats were exposed to HBH equivalent to 6,100 m for 7 days in a simulated decompression chamber. After stipulated period of exposure, learning ability was assessed using Morris water maze (MWM) task. Cholinergic markers like acetylcholine (ACh) and acetyl cholinesterase (AChE) were evaluated from cortex and hippocampus. Morphological changes were evaluated from cortex, CA1, and CA3 region of hippocampus by Nissle staining and by electron microscopy. We found that exposure to HBH led to impairment of learning ability in MWM task, and it was accompanied by decrease in ACh level, increase in AChE activity, and revealed critical cellular damage. Administration of AChEIs like physostigmine (PHY) and galantamine (GAL) resulted in amelioration of the deleterious effects induced by HBH. The AChEIs were also able to restore the neuronal morphology. Our data suggest that cholinergic system is affected by HBH, and AChEIs were able to improve HBH-induced learning impairment in rats.     

Suppression of IgE responses in inbred rats by repeated respiratory tract exposure to antigen: responder phenotype influences isotype specificity of induced tolerance

Repeated exposure of rats to an aerosol of ovalbumin (OVA) induced tolerance to subsequent parenteral challenge with the same antigen. In the low-IgE responder WAG strain, responses in both the IgE and IgG classes were affected, whereas rats of the moderate (Lou/M) and high-IgE responder BN strain developed high titers of anti-OVA IgG in serum during exposure with concomitant tolerance in the IgE class. Repeated parenteral challenge, however, failed to elicit significant secondary anti-OVA IgG responses in the Lou/M and BN strains, suggesting that the isotype specificity of induced tolerance in these strains was not absolute. Spleen and respiratory tract lymph node cells, but not serum from aerosol-exposed BN rats, were capable of transferring IgE isotype- and antigen-specific tolerance. Dose response experiments demonstrated that the low-IgE responder WAG strain was exquisitely sensitive to tolerance induction in response to antigen inhalation, being susceptible to dosages in the nanogram range; at least 1000 times more antigen was required in the high-IgE responder BN to induce comparable tolerance in the IgE class. It was also apparent that the IgE isotype was more readily suppressed than the IgG isotype in both high- and low-IgE responder strains.     

Effects of water deprivation and prior LiCl exposure in conditioning taste aversions

The phenomenon of attenuated taste aversion conditioning following preexposure to a lithium chloride UCS was demonstrated in Experiment 1. The amount of fluid consumed during the preconditioning phase was shown to be a factor in the degree of attenuated conditioning. The pharmacological properties of LiCl, and the effects of either ad lib or restricted fluid intake in conjunction with state dependent variables were proposed to account for the results. In Experiment 2 the water scheduling and LiCl habituation procedures of Experiment 1 were replicated, and serum lithium was assessed in the various groups on conditioning day. The groups did not differ, ruling out an incremental illness hypothesis of attenuated conditioning.