Craig J. Slawecki

Periadolescent alcohol exposure has lasting effects on adult neurophysiological function in rats

Most individuals have their first experience with ethanol (EtOH) consumption as adolescents. Episodes of high EtOH drinking, lasting from hours to days (i.e. binges), are not uncommon. Thus, adolescent EtOH drinking has become a significant health concern due to the possible protracted effects of high doses of EtOH on behavior and the developing brain. This study assessed the effects of brief high levels of EtOH during periadolescence on subsequent behavior and electrophysiology in adult rats. Male Sprague-Dawley rats were exposed to EtOH vapor for 5 days (i.e. postnatal days 35-40) or 10 days (i.e. postnatal days 30-40) for 12 h/day. Locomotor activity, EEG activity, and event-related potentials (ERPs) were then assessed at 1 and 6-7 weeks post EtOH exposure. Significant differences in locomotor activity were not observed at 1 week or 6-7 weeks post-ethanol exposure. However, EtOH exposure did have long-term electrophysiological effects. EtOH exposure increased the frequency of the EEG in the 1-2 Hz range in the parietal cortex and the 16-32 Hz range in the hippocampus. EtOH exposure also increased hippocampal N2 amplitude, decreased hippocampal P3 amplitude, and decreased cortical and hippocampal P2 amplitudes. While these findings are generally similar to those reported following long-term ethanol exposure during adulthood, alcohol exposure during adolescence appears to produce more robust hippocampal effects following shorter periods of exposure. In addition, these data indicate that, in the absence of overt behavioral differences, there are long-lasting changes in the functional brain activity of adult rats briefly exposed to high levels of EtOH during the periadolescent period.

The effects of social isolation on neuropeptide Y levels, exploratory and anxiety-related behaviors in rats

NPY is one of the most abundantly expressed peptides within the CNS, and has been previously demonstrated to be altered in several animal models of depression, as well as to be differentially regulated by acute and repeated stress. The effect of social deprivation, through isolation housing, on brain NPY concentrations in adult rats has not been previously investigated. The effects of 12 weeks of social isolation, in adult rats, on anxiety-related behaviors and central concentrations of NPY in: hypothalamus, amygdala, caudate-putamen, hippocampus, and frontal cortex were evaluated. Single housed animals spent significantly more time on the open arms of the elevated plus maze and in the central area of the open field as compared to pair housed controls. These data are most likely indicative of enhanced exploration and novelty seeking. Concentrations of neuropeptide Y were increased in the caudate-putamen of the single housed subjects. NPY levels in caudate/putamen and hypothalamus were also significantly correlated with time spent in the open arms of the elevated plus maze. These data suggest that chronic social isolation, in these adult Wistar rats, did not increase anxiety but produced enhanced exploration in tests of anxiety, an effect that was associated with NPY concentrations in the striatum and hypothalamus.

Increased anxiety-like behavior in adult rats exposed to nicotine as adolescents

OBJECTIVE Twenty percent of adolescents between 12 and 18 years old are regular smokers. Recently developed animal models demonstrate that adolescent nicotine exposure produces behavioral and electrophysiological changes, which persist into adulthood. The purpose of this study was to further define the behavioral effects of nicotine exposure during adolescence. METHODS Male 31-36-day-old adolescent rats were administered 5.0 mg/kg/day nicotine using transdermal Nicoderm CQ patches (SmithKline Beecham). During nicotine exposure, motor activity was assessed. Behavior in both standard open field and modified open field was examined 2-3 weeks after exposure ended. RESULTS Nicotine exposure significantly enhanced motor activity in nicotine-exposed rats compared with controls, demonstrating the acute stimulatory effects of transdermal nicotine. Two to three weeks after nicotine exposure ended, significantly lower levels of exploratory activity were observed relative to controls in the standard open field. Rats exposed to nicotine during adolescence also retreated to the perimeter of the open field more quickly than control rats. In a modified open field, nicotine exposure reduced approaches to food, contact with food and food intake. CONCLUSIONS Taken together, these data suggest that adolescent nicotine exposure may induce an anxiogenic profile, which persists beyond acute nicotine withdrawal. Given the hypothesized role of stress and anxiety in the maintenance of smoking, it could be speculated that anxiety associated with smoking abstinence may play an important role in continued adolescent tobacco use.

Increases in sucrose consumption, but not ethanol consumption, following ICV NPY administration

Neuropeptide Y (NPY) is a centrally acting neuromodulator that influences both consummatory behaviors and anxiety. NPYs effects on feeding are primarily regulated through Y5 receptors in hypothalamic sites, whereas NPY-induced anxiolysis appears to be mediated by Y1 receptors in the amygdala. Recently, NPY has been postulated to play a role in the regulation of ethanol consumption. The present study assessed the influence of intracerebroventricular (ICV) administration of NPY on the consumption of 10% ethanol or 2% sucrose in rats. Male Wistar rats were trained to self-administer 10% ethanol using the sucrose-substitution procedure and then implanted with an intracerebroventricular (ICV) cannula. The effects of NPY (0-15 microg) on ethanol consumption and sucrose consumption were then examined. ICV NPY infusion had no significant effects on the consumption of 10% ethanol, however, NPY significantly increased the consumption of 2% sucrose, [F(1, 11) = 6.18, p = 0.03]. These data suggest that ethanol intake and sucrose intake are differentially regulated by NPY. It is hypothesized that ICV infusion of NPY may be affecting both Y1 and Y5 receptors producing increased consummatory drive and anxiolysis, two factors that have opposing effects on subsequent ethanol consumption. Therefore, additional studies including site specific injection of NPY will be necessary to provide further insight into the role of NPY on ethanol consumption.

Effects of adolescent ethanol exposure on ethanol consumption in adult rats

Ethanol exposure during early development could predispose an individual to increased ethanol consumption. Given the high prevalence of adolescent ethanol abuse, it is important to assess the potential impact of adolescent ethanol exposure on the development of alcohol drinking. The following study was designed to assess the initiation of ethanol consumption in adult rats after exposure to ethanol vapors during adolescence. Male Sprague-Dawley rats (n = 23) were exposed to ethanol vapor for 12 h per day for 10 consecutive days between postnatal days 30 and 40. Ethanol vapor exposure maintained blood ethanol levels averaging 250 mg/dl. All rats were subsequently trained to self-administer ethanol after a 52-day withdrawal period. When ethanol consumption was assessed in the adult rats (>3 months old) there were no significant differences in initiation or maintenance of ethanol self-administration between ethanol-exposed and control rats. In addition, there were no group differences in the ability of a noise stressor presented before the drinking session to transiently decrease ethanol intake. Overall, these findings indicate that forced exposure to ethanol vapor during adolescence does not seem to be sufficient to alter initiation or maintenance of limited-access ethanol self-administration.

Effects of chronic ethanol exposure on sleep in rats

Sleep disturbance is a common complaint in alcoholics. When polysomnographic studies are performed in alcoholics, reductions in slow wave sleep are a common finding; however, few studies have evaluated the effects of chronic alcohol exposure on sleep in animal models. In the present study, the sleep EEG was evaluated in 40 Wistar rats who were exposed to chronic alcohol or control conditions in vapor chambers. Rats were exposed to ethanol vapors or control chambers for 6 weeks and then withdrawn. Sleep EEG was recorded before exposure (baseline), immediately following exposure, and 5 weeks after withdrawal from the ethanol/control chambers. In the ethanol-exposed animals, blood ethanol levels averaged 192 mg/dL over 6 weeks of exposure. Chronic ethanol exposure and withdrawal was not found to affect either slow wave sleep latency or slow wave sleep duration; however, overall spectral power as well as power in the delta, theta, and beta frequencies were significantly reduced following chronic exposure (2-4 Hz, [F(1, 17) = 18.11, p = 0.001], 4-6 Hz, [F(1, 17) = 15.98, p = 0.001], 6-8 Hz [F(1, 17) = 15.52, p = 0.001], 8-16 Hz band [F(1, 17) = 18.73, p < 0.0001], 16-32 Hz [F(1, 17) = 10.13, p = 0.005], and 1-50 Hz [F(1, 17) = 17.03, p = 0.001]. After 5 weeks of withdrawal, significant decreases still persisted in the delta and theta frequencies (2-4 Hz [F(1, 16) = 6.21, 0.024], 4-6 Hz [F(1, 16) = 6.26, 0.024], and 6-8 Hz [F(1, 16) = 4.84, p = 0.043]). These findings suggest that spectral analysis of the EEG is a highly sensitive measure of the effects of ethanol on sleep. These findings additionally demonstrate that chronic ethanol exposure can produce persistent diminution in the systems that generate cortical slow waves in the rat and thus may provide a model for understanding the mechanisms underlying sleep disturbances associated with alcoholism.

Nicotine exposure during the neonatal brain growth spurt produces hyperactivity in preweanling rats

Despite warning labels and increases in evidence of the adverse effects of tobacco use, women continue to use tobacco products during pregnancy. Cigarette smoking has been linked to increased prenatal mortality, increased incidence of SIDS, reductions in birth weight, and disruptions in CNS and behavioral development. Animal model systems have critically established the causal relationship between nicotine and adverse developmental outcome. The present study examines the behavioral effects of nicotine exposure in the rat during the third trimester equivalent of the human brain growth spurt, a period of rapid development of the cholinergic systems and a period during which the CNS is particularly vulnerable to a number of insults. Sprague-Dawley rat pups were exposed to nicotine (6.0 mg/kg/day) from postnatal days (PD) 4-9 via an artificial rearing procedure. This procedure ensures that observed effects are not due to nutritional deficits. Two control groups were employed, an artificially reared control group and a normally reared control group. Activity level was measured on PD 18-19. Nicotine-exposed subjects were significantly overactive compared to both control groups, which did not differ significantly from one another. This behavioral alteration was observed in the absence of nicotine-induced body weight deficits. These results suggest that women who use tobacco products during late gestation may place their fetuses at risk for hyperactivity later in life, particularly during early adolescence.

Increased CRF-like and NPY-like immunoreactivity in adult rats exposed to nicotine during adolescence: Relation to anxiety-like and depressive-like behavior

OBJECTIVE Recently, animal models have been developed that demonstrate that adolescent nicotine exposure produces neurobehavioral changes which persist into adulthood. This study further examined the impact of adolescent nicotine exposure on anxiety-like and depressive-like behavior, as well as on levels of corticotropin-releasing factor (CRF) and neuropeptide Y (NPY) in this model. METHODS Male adolescent rats (35-40 days old) were administered nicotine using Nicoderm CQ patches (Smith-Kline Beecham). Behavior in the elevated plus maze (EPM) and forced swim test (FST) was assessed 2-3 weeks after exposure ended. Brain levels of CRF and NPY were then assessed 5-6 weeks after behavioral tests were completed. In addition, blood and brain levels of nicotine resulting from nicotine treatment were examined. RESULTS After 5 days of exposure to 5 mg/kg/day nicotine, blood levels of nicotine averaged 66+/-5 ng/ml and brain nicotine levels averaged 52+/-4 ng/g. Rats exposed to nicotine displayed an anxiety-like profile in the EPM (i.e., decreased time spent in the open arms) and an antidepressant-like profile in the FST (i.e., less time spent immobile). Rats exposed to nicotine also had increased hypothalamic and frontal cortical CRF, increased hypothalamic and hippocampal NPY, and a decreased ratio of NPY to CRF in the amygdala. CONCLUSIONS This study demonstrates that adolescent nicotine exposure produces lasting increases in anxiety-like behavior and may reduce depressive-like behavior. These behavioral changes also occurred in concert with alterations in CRF and NPY systems. Thus, lasting neurobehavioral changes associated with adolescent nicotine exposure may be related to allostatic changes in stress peptide systems.

Increased alcohol drinking in isolate-housed alcohol-preferring rats.

Alcoholism is a complex disorder influenced by interactions between genetic and environmental risk factors. This study examined the influence of isolate housing on ethanol intake in alcohol-preferring (P) and non-alcohol-preferring (NP) rats. Rats were isolate-housed or pair-housed for 8 weeks when between 45 and 96 days old. Ethanol drinking was assessed using a 24-hr preference test (10% ethanol vs. water) and 20-min limited access tests. A behavioral test battery was used to assess anxiety-like, depressive-like, acoustic startle, and motor behavior. Isolate housing increased home cage drinking in both lines and increased limited access drinking selectively in P rats. Isolation also reduced swim test immobility and prepulse inhibition in P rats and increased locomotor stereotypies in NP rats. Taken together, these data demonstrate that LinexEnvironment interactions influence the effects of isolation. Furthermore, isolation selectively increased ethanol intake in high drinking P rats. This effect was not correlated with changes in other behaviors. Selective enhancement of limited access ethanol drinking in P rats may represent a model whereby genetic liability to excessive drinking is enhanced by specific environmental exposures.

Comparison of anxiety-like behavior in adolescent and adult sprague-dawley rats

The conditions under which age differences in anxiety are observed in rodents are unclear. These studies explored the influence of test condition on anxiety-like behavior in adolescent and adult rats using the light-dark box. Behavior was assessed under different illumination levels (30 or 60 lux) and after brief stress (restraint or bright light). Anxiety-like behavior did not differ in the 30-lux test but was increased in adolescents in the 60-lux test. Restraint increased anxiety-like behavior in adolescents, resulting in elevated anxiety-like behavior relative to adults. Bright light decreased anxiety-like behavior, possibly because of negative contrast or novelty-induced anxiolysis. These studies demonstrate that adolescents display increased anxiety-like behavior when test conditions are more aversive and following exposure to stress.