Jaime L. Diaz-Granados

The role of GABAA receptors in the acute and chronic effects of ethanol: a decade of progress

The past decade has brought many advances in our understanding of GABAA receptor-mediated ethanol action in the central nervous system. We now know that specific GABAA receptor subtypes are sensitive to ethanol at doses attained during social drinking while other subtypes respond to ethanol at doses attained by severe intoxication. Furthermore, ethanol increases GABAergic neurotransmission through indirect effects, including the elevation of endogenous GABAergic neuroactive steroids, presynaptic release of GABA, and dephosphorylation of GABAA receptors promoting increases in GABA sensitivity. Ethanol’s effects on intracellular signaling also influence GABAergic transmission in multiple ways that vary across brain regions and cell types. The effects of chronic ethanol administration are influenced by adaptations in GABAA receptor function, expression, trafficking, and subcellular localization that contribute to ethanol tolerance, dependence, and withdrawal hyperexcitability. Adolescents exhibit altered sensitivity to ethanol actions, the tendency for higher drinking and longer lasting GABAergic adaptations to chronic ethanol administration. The elucidation of the mechanisms that underlie adaptations to ethanol exposure are leading to a better understanding of the regulation of inhibitory transmission and new targets for therapies to support recovery from ethanol withdrawal and alcoholism.

Behavioral effects of ethanol in cerebellum are age dependent: potential system and molecular mechanisms.

BACKGROUND Adolescent rats are less sensitive to the motor-impairing effects of ethanol than adults. However, the cellular and molecular mechanisms underlying this age-dependent effect of ethanol have yet to be fully elucidated. METHOD Male rats of various ages were used to investigate ethanol-induced ataxia and its underlying cellular correlates. In addition, Purkinje neurons from adolescent and adult rats were recorded both in vivo and in vitro. Finally, protein kinase C (PKCγ) expression was determined in 3 brain regions in both adolescent and adult rats. RESULTS The present multi-methodological investigation confirms that adolescents are less sensitive to the motor-impairing effects of ethanol, and this differential effect is not because of differential blood ethanol levels. In addition, we identify a particular cellular correlate that may underlie the reduced motor impairment. Specifically, the in vivo firing rate of cerebellar Purkinje neurons recorded from adolescent rats was insensitive to an acute ethanol challenge, while the firing rate of adult cerebellar Purkinje neurons was significantly depressed. Finally, it is demonstrated that PKCγ expression in the cortex and cerebellum mirrors the age-dependent effect of ethanol: adolescents have significantly less PKCγ expression compared to adults. CONCLUSIONS Adolescents are less sensitive than adults to the motor-impairing effects of ethanol, and a similar effect is seen with in vivo electrophysiological recordings of cerebellar Purkinje neurons. While still under investigation, PKCγ expression mirrors the age effect of ethanol and may contribute to the age-dependent differences in the ataxic effects of ethanol.

Alcohol use across the lifespan: An analysis of adolescent and aged rodents and humans.

Adolescence and old age are unique periods of the lifespan characterized by differential sensitivity to the effects of alcohol. Adolescents and the elderly appear to be more vulnerable to many of alcohols physiological and behavioral effects compared to adults. The current review explores the differential effects of acute alcohol, predominantly in terms of motor function and cognition, in adolescent and aged humans and rodents. Adolescents are less sensitive to the sedative-hypnotic, anxiolytic, and motor-impairing effects of acute alcohol, but research results are less consistent as it relates to alcohols effects on cognition. Specifically, previous research has shown adolescents to be more, less, and similarly sensitive to alcohol-induced cognitive deficits compared to adults. These equivocal findings suggest that learning acquisition may be differentially affected by ethanol compared to memory, or that ethanol-induced cognitive deficits are task-dependent. Older rodents appear to be particularly vulnerable to the motor- and cognitive-impairing effects of acute alcohol relative to younger adults. Given that alcohol consumption and abuse is prevalent throughout the lifespan, it is important to recognize age-related differences in response to acute and long-term alcohol. Unfortunately, diagnostic measures and treatment options for alcohol dependence are rarely dedicated to adolescent and aging populations. As discussed, although much scientific advancement has been made regarding the differential effects of alcohol between adolescents and adults, research with the aged is underrepresented. Future researchers should be aware that adolescents and the aged are uniquely affected by alcohol and should continue to investigate alcohols effects at different stages of maturation.

The effect of chronic intermittent ethanol exposure on spatial memory in adolescent rats: The dissociation of metabolic and cognitive tolerances

Using a rapid chronic intermittent ethanol (CIE) vapor exposure paradigm, we demonstrate the dissociability of metabolic tolerance from cognitive tolerance in adolescent rats. Adolescent rats were trained to spatially navigate in the Morris Water Maze and then exposed to CIE vapor or air 16 h a day for 4 days. After a final 28 h withdrawal, all rats received a saline or ethanol challenge, followed by a test of spatial memory 30 min after administration. Results indicate that CIE vapor exposure did not significantly impair adolescent spatial memory. Although CIE-exposed rats developed metabolic tolerance to a subsequent ethanol administration, CIE exposure did not alter dose-dependent ethanol-induced spatial memory impairments. These data indicate that metabolic ethanol tolerance can be distinguished from cognitive ethanol tolerance during adolescence and suggest that blood alcohol levels alone do not fully explain ethanol-induced spatial memory impairments.

Periadolescent exposure to ethanol and diazepam alters the aversive properties of ethanol in adult mice

Evidence suggests that the developing adolescent brain may be especially vulnerable to long-term neurobehavioral consequences following ethanol exposure and withdrawal. In the present study, we examined the long-term effect of adolescent ethanol withdrawal on a subsequent EtOH-induced conditioned taste aversion (CTA). Periadolescent and adult C3H mice were exposed to 64 h of continuous (single withdrawal) or intermittent (multiple withdrawal) ethanol vapor. Following each ethanol exposure, animals received either 0, 1, 2, or 3 mg/kg diazepam (DZP) in an attempt to counteract the possible effect of ethanol withdrawal. About 6 weeks following ethanol and DZP treatment, animals were tested for an EtOH-induced CTA. As expected, exposure to EtOH during adolescence attenuated the EtOH-induced CTA as compared to controls. Unexpectedly, administration of DZP during withdrawal did not spare but rather mimicked the attenuation of the EtOH-induced CTA seen in animals exposed to ethanol in adolescence. This attenuation was not evident when EtOH and/or DZP was administered in adulthood. Given the similar mode of action of EtOH and DZP on the GABA system, the principal implication of the present findings is that the intoxicating effect of ethanol on the developing brain can result in long-term changes in the aversive properties of EtOH.

Low and moderate doses of acute ethanol do not impair spatial cognition but facilitate accelerating rotarod performance in adolescent and adult rats

Adolescents and adult rodents have differing sensitivities to the acute effects of ethanol on a variety of behavioral and electrophysiological measures. Often, these differences are revealed using high ethanol doses and consequently little is known about these age-related effects using lower ethanol doses. We sought to determine if low-dose ethanol produces differential effects on cognition and motor behavior in adolescent and adult rats. Adolescent (postnatal day PD 30-32) and adult (PD 70-72) male Sprague Dawley rats were trained on the standard version of the Morris Water Maze (MWM) for 5 days or received 5 training trials on an accelerating rotarod (ARR). Adolescents learned the location of the submerged platform in the MWM significantly slower than adults during training and, acute ethanol administration (0.5 g/kg, 0.75 g/kg, or 1.0 g/kg) 30 min before testing did not impair spatial memory in either age group. On the ARR test, adolescent rats spent significantly more time on the rotarod compared to adults and, alcohol exposure (1.0 g/kg) significantly increased ARR performance 30 min following administration. Our findings address the utility of investigating low and moderate doses of ethanol during different developmental stages in rats.

The Influence of a Chronic Adolescent Nicotine Exposure on Ethanol Withdrawal Severity During Adulthood in C3H Mice

Animal and human studies have shown tolerance, consumption, relapse, and behavioral interactions between ethanol and nicotine, but little is understood about their interaction, especially as it relates to ethanol withdrawal in adulthood for subjects who have an adolescent history of using these drugs. This study investigated nicotines influence on ethanol withdrawal seizures in two different age groups of male C3H mice. Adolescent and adult male C3H mice, beginning at postnatal day 28 or 70, respectively, were subjected to a 7-day chronic exposure to ethanol only, ethanol plus nicotine, nicotine only, or vehicle treatment. Six weeks later, all the groups were subjected to chronic exposure to ethanol vapors and the severity of their ethanol withdrawal seizures was assessed by handling-induced convulsions. An adolescent exposure to chronic nicotine resulted in an exacerbation of ethanol withdrawal seizures in adulthood. Given this, adolescence may contain a neurophysiological critical period that is sensitive to nicotine and which may result in an altered response to ethanol dependency in adulthood. These findings have serious implications for the long-term consequences following co-abuse of these drugs during adolescence.

Investigation of ethanol-induced impairment of spatial memory in γ2 heterozygous knockout mice

GABA(A) receptors, the major inhibitory receptors in the mammalian central nervous system, are affected by a number of drug compounds, including ethanol. The pharmacological effects of certain drugs have been shown to be dependent upon specific GABA(A) receptor subunits. Because benzodiazepines and ethanol have similar effect signatures, it has been hypothesized that these drugs share the gamma2-containing GABA(A) receptors as a mechanism of action. To probe the involvement of the gamma2 subunit in ethanols actions, spatial memory for the Morris water maze task was tested in gamma2 heterozygous knockout mice and wild type littermate controls following ethanol administration at the following doses: 0.0, 1.25, 1.75, and 2.25 g/kg. While baseline learning and memory were unaffected by reduction of gamma2 containing GABA(A) receptors, ethanol dose-dependently impaired spatial memory equally in gamma2 heterozygous knockouts and wild type littermate controls.

Impact of adolescent alcohol use across the lifespan: Long-lasting tolerance to high-dose alcohol coupled with potentiated spatial memory impairments to moderate-dose alcohol

Understanding how alcohol exposure during adolescence affects aging is a critical but understudied area. In the present study, male rats were exposed to either alcohol or saline during adolescence, then tested every 4 months following either an ethanol or saline challenge; animals were tested until postnatal day (PD) 532. It was found that long-lasting tolerance to high-dose ethanol exists through the test period, as measured by loss of righting reflex, while tolerance to lower doses of ethanol is not found. In addition, alcohol exposure during adolescence facilitated spatial memory impairments to acute ethanol challenges later in life. The current work demonstrates that exposure to ethanol during adolescent development can produce long-lasting detrimental impairments.