Exercise reduces physical alterations in a rat model of fetal alcohol spectrum disorders.

BACKGROUND: Prenatal alcohol exposure (PAE) has serious physical consequences for children such as behavioral disabilities, growth disorders, neuromuscular problems, impaired motor coordination, and decreased muscle tone. However, it is not known whether loss of muscle strength occurs, and which interventions will effectively mitigate physical PAE impairments. We aimed to investigate whether physical alteration persists during adolescence and whether exercise is an effective intervention. RESULTS: Using paradigms to evaluate different physical qualities, we described that early adolescent PAE animals have significant alterations in agility and strength, without alterations in balance and coordination compared to CTRL animals. We evaluated the effectiveness of 3 different exercise protocols for 4 weeks: Enrichment environment (EE), Endurance exercise (EEX), and Resistance exercise (REX). The enriched environment significantly improved the strength in the PAE group but not in the CTRL group whose strength parameters were maintained even during exercise. Resistance exercise showed the greatest benefits in gaining strength, and endurance exercise did not. CONCLUSION: PAE induced a significant decrease in strength compared to CTRL in PND21. Resistance exercise is the most effective to reverse the effects of PAE on muscular strength. Our data suggests that individualized, scheduled, and supervised training of resistance is more beneficial than endurance or enriched environment exercise for adolescents FASD.

Bioinformatic analysis predicts that ethanol exposure during early development causes alternative splicing alterations of genes involved in RNA post-transcriptional regulation.

Prenatal ethanol exposure is associated with neurodevelopmental defects and long-lasting cognitive deficits, which are grouped as fetal alcohol spectrum disorders (FASD). The molecular mechanisms underlying FASD are incompletely characterized. Alternative splicing, including the insertion of microexons (exons of less than 30 nucleotides in length), is highly prevalent in the nervous system. However, whether ethanol exposure can have acute or chronic deleterious effects in this process is poorly understood. In this work, we used the bioinformatic tools VAST-TOOLS, rMATS, MAJIQ, and MicroExonator to predict alternative splicing events affected by ethanol from available RNA sequencing data. Experimental protocols of ethanol exposure included human cortical tissue development, human embryoid body differentiation, and mouse development. We found common genes with predicted differential alternative splicing using distinct bioinformatic tools in different experimental designs. Notably, Gene Ontology and KEGG analysis revealed that the alternative splicing of genes related to RNA processing and protein synthesis was commonly affected in the different ethanol exposure schemes. In addition, the inclusion of microexons was also affected by ethanol. This bioinformatic analysis provides a reliable list of candidate genes whose splicing is affected by ethanol during nervous system development. Furthermore, our results suggest that ethanol particularly modifies the alternative splicing of genes related to post-transcriptional regulation, which probably affects neuronal proteome complexity and brain function.

Changes in the clustering of health-related behaviors during the COVID-19 pandemic: examining predictors using latent transition analysis.

The COVID-19 pandemic has had a significant impact on daily life, affecting both physical and mental health. Changes arising from the pandemic may longitudinally impact health-related behaviors (HRB). As different HRBs co-occur, in this study, we explore how six HRBs - alcohol (past-week and binge-drinking), tobacco, marijuana, benzodiazepine use, and unhealthy food consumption - were grouped and changed over time during the COVID-19 pandemic. A sample of 1038 university students and staff (18 to 73 years old) of two universities completed an online psychometrically adequate survey regarding their recalled HRB (T0, pre-COVID-19 pandemic) and the impact of COVID-19 on their behaviors during July (T1) and November (T2). Latent Transition Analysis (LTA) was used to identify HRB cluster membership and how clusters changed across T0, T1, and T2. Four clusters emerged, but remained mainly stable over time: 'Lower risk' (65.2-80%), 'Smokers and drinkers' (1.5-0.01%), 'Binge-drinkers and marijuana users' (27.6-13.9%), and 'Smokers and binge-drinkers' (5.6-5.8%). Participants who moved from one cluster to another lowered their HRB across time, migrating from the 'Binge-drinkers and marijuana users' cluster to 'Lower risk'. Participants in this cluster were characterized as less affected economically by the COVID-19 pandemic, with lower reported stress levels, anxiety, depression, and loneliness than the other clusters. Our results provide evidence of how HRBs clustered together and transitioned longitudinally during the COVID-19 pandemic. HRB clustering across time offers a valuable piece of information for the tailoring of interventions to improve HRB.

The Perceived Impact of COVID-19 on Comfort Food Consumption over Time: The Mediational Role of Emotional Distress.

The COVID-19 pandemic has had a significant impact on populations at an economic, health, and on an interpersonal level, it is still unclear how it has affected health-risk behaviors, such as comfort food consumption over time. This study longitudinally examines the effect of the perceived impact of COVID-19 on comfort food consumption and whether this effect is mediated by emotional distress. A convenience sample of 1048 students and university staff (academic and non-academic) from two universities completed monthly online surveys during the COVID-19 pandemic across six waves (W; W1 to W6). Participants reported their perceived impact of COVID-19 (economic, interpersonal, and health), comfort food consumption, and emotional distress (DASS-21). Using structural equation models, we found an indirect longitudinal effect of the perceived impact of COVID-19 (W1) on comfort food consumption (W3 to W6) through increased emotional distress (W2). The perceived negative impact of COVID-19 on comfort food consumption was fully mediated by the emotional distress during the first waves (W3 and W4), ending in a partial mediation in the last waves (W5 and W6). These findings contribute to disentangling the mechanisms by which the perceived impact of COVID-19 affects comfort food consumption over time, and highlight the role of emotional distress. Future interventions should address comfort food consumption by focusing on handling emotional distress during a crisis.

COVID-19 Perceived Impact and Psychological Variables as Predictors of Unhealthy Food and Alcohol Consumption Trajectories: The Role of Gender and Living with Children as Moderators.

The present study examines the trajectories of unhealthy food and alcohol consumption over time and considers whether perceived impact of COVID-19 and psychological variables are predictors of these trajectories. We ascertained whether these predictors are different in women vs. men and between women living with vs. without children. Data were collected through online surveys administered to 1038 participants from two universities (staff and students) in Chile, across five waves (July to October 2020). Participants provided information about their past-week unhealthy food and alcohol consumption and mental health. Using latent growth curve modeling analysis, we found that higher perceived health and interpersonal COVID-19 impact, younger age and lower depression symptoms were associated with more rapid increases over time in unhealthy food consumption. On the other hand, higher perceived COVID-19 economic impact and older age were associated with more rapid diachronic decreases in alcohol consumption. Gender and living with or without children, for women only, were moderators of these trajectories. This longitudinal study provides strong evidence identifying the multiple repercussions of COVID-19 and mental health factors on unhealthy food and alcohol consumption. These findings highlight the need for interventions aimed at minimizing the impact of the pandemic on unhealthy food and alcohol consumption over time.

N-Methyl-d-Aspartate Receptor Modulation by Nicotinamide Adenine Dinucleotide Phosphate Oxidase Type 2 Drives Synaptic Plasticity and Spatial Memory Impairments in Rats Exposed Pre- and Postnatally to Ethanol.

Aims: Pre- and/or early postnatal ethanol exposure (prenatal alcohol exposure [PAE]) impairs synaptic plasticity as well as memory formation, but the mechanisms underlying these effects remain unclear. Both long-term potentiation (LTP) and spatial memory formation in the hippocampus involve the nicotinamide adenine dinucleotide phosphate oxidase type 2 (NOX2) enzyme. Previous studies have reported that N-methyl-d-aspartate receptor (NMDAR) activation increases NOX2-mediated superoxide generation, resulting in inhibition of NMDAR function, but whether NOX2 impacts NMDAR function in PAE animals leading to impaired LTP and memory formation remains unknown. We aim to evaluate whether the NOX2-NMDAR complex is involved in the long-lasting deleterious effects of PAE on hippocampal LTP and memory formation. Results: Here we provide novel evidence that PAE animals display impaired NMDAR-dependent LTP in the cornus ammonis field 1 (CA1) and NMDAR-mediated LTP in the dentate gyrus (DG). Moreover, PAE rats displayed increased NMDAR-mediated transmission in both hippocampal areas. Interestingly, NOX2 pharmacological inhibition restored NMDAR-mediated transmission and LTP in the CA1, but not in the DG. PAE also induced overexpression of NOX2 and CaMKII isoforms, but did not modify the content or the redox state of the N-methyl-d-aspartate receptor subunit-1 (NR1) subunit of NMDAR in both areas of the hippocampus. In addition, adolescent PAE rats orally fed the antioxidant and free radical scavenger apocynin exhibited significantly improved spatial memory acquisition. Innovation and Conclusion: By showing in PAE animals NOX2 overexpression and increased NMDAR-mediated transmission, which might lead to impaired synaptic plasticity and memory formation in a region-specific manner, we provide an important advance to our current understanding of the cellular mechanisms underlying PAE-dependent defective hippocampal function.

Pre- and postnatal alcohol exposure delays, in female but not in male rats, the extinction of an auditory fear conditioned memory and increases alcohol consumption.

Repeated exposure to alcohol increases retrieval of fear-conditioned memories, which facilitates, among other factors, the emergence of post-traumatic stress disorder (PTSD). Individuals with PTSD are more likely to develop alcohol and substance abuse related disorders. We assessed if prenatal and early postnatal alcohol exposure (PAE) increased the susceptibility to retain aversive memories and if this was associated with subsequent heightened alcohol consumption. Pregnant Sprague-Dawley rats were exposed for 22 hr/day, throughout pregnancy and until postnatal Day 7 to a single bottle of sucralose - sweetened 10% alcohol solution (PAE Group), or to a single bottle of tap water and sucralose (Control Group). Auditory fear conditioning (AFC) was performed in the adolescent offspring at postnatal Day 40. Freezing was measured during acquisition, retention and extinction phases, followed by 3 weeks of free choice alcohol intake. Female, but not male, PAE rats exhibited impaired extinction of the aversive memory, a finding associated with higher levels of 3-4 Dihidroxyphenylacetic acid (DOPAC) in the nucleus accumbens and heightened alcohol intake, respect to controls. These findings suggest that PAE makes females more vulnerable to long-term retention of aversive memories, which coexist with heightened alcohol intake. These findings are reminiscent of those of PTSD.

NADPH Oxidase Isoform 2 (NOX2) Is Involved in Drug Addiction Vulnerability in Progeny Developmentally Exposed to Ethanol.

Ethanol exposure increases oxidative stress in developing organs, including the brain. Antioxidant treatment during maternal ethanol ingestion improves behavioral deficits in rodent models of fetal alcohol spectrum disorder (FASD). However, the impact of general antioxidant treatment in their adult offspring and the Specific Reactive Species (ROS)-dependent mechanism, are not fully understood. We hypothesized that pre and early postnatal ethanol exposure (PEE) modifies redox homeostasis, in particular NOX2 function during reward signaling in the mesocorticolimbic pathway, which reinforces the effects of alcohol. We developed a FASD rat model which was evaluated during adolescence (P21) and adulthood (P70). We first studied whether redox homeostasis is affected in PEE animals, by analyzing mRNA expression of SOD1, CAT, and Gpx1. We found that PEE reduced the mRNA levels of these three anti-oxidant enzymes in PFC and HIPP at P21 and in the VTA at P70. We also analyzed basal mRNA and protein expression of NOX2 subunits such as gp91phox, p22 phox, and p47 phox, in mesocorticolimbic brain areas of PEE rat brains. At P21, gp91 phox, and p47 phox levels in the VTA were decreased. At P70, gp91 phox mRNA levels was decreased in HIPP and both mRNA and protein levels were decreased in PFC. Since NOX2 is regulated by the N-methyl-D-aspartate Receptor (NMDAR), we analyzed NMDAR mRNA expression and found differential expression of NMDAR subunits (NR1 and NR2B) in the PFC that was age dependent, with levels decreased at P21 and increased at P70. The analysis also revealed decreased NR2B mRNA expression in HIPP and VTA at P70. Offspring from maternal ethanol users consumed 25% more ethanol in a free choice alcohol consumption test than control rats, and showed place preference for an alcohol-paired compartment. In vivo inhibition of NOX2 using apocynin in drinking water, or infusion of blocked peptide gp91 phox ds in the VTA normalized alcohol place preference, suggesting that NOX2 plays an important role in addictive like behavior. Taken together, PEE significantly affects the expression of antioxidant enzymes, NOX2, NMDAR in an age, and brain region dependent manner. Moreover, we demonstrate that NOX2 regulates alcohol seeking behavior.

Neuroligin1 drives synaptic and behavioral maturation through intracellular interactions.

In vitro studies suggest that the intracellular C terminus of Neuroligin1 (NL1) could play a central role in the maturation of excitatory synapses. However, it is unknown how this activity affects synapses in vivo, and whether it may impact the development of complex behaviors. To determine how NL1 influences the state of glutamatergic synapses in vivo, we compared the synaptic and behavioral phenotypes of mice overexpressing a full-length version of NL1 (NL1FL) with mice overexpressing a version missing part of the intracellular domain (NL1ΔC). We show that overexpression of full-length NL1 yielded an increase in the proportion of synapses with mature characteristics and impaired learning and flexibility. In contrast, the overexpression of NL1ΔC increased the number of excitatory postsynaptic structures and led to enhanced flexibility in mnemonic and social behaviors. Transient overexpression of NL1FL revealed that elevated levels are not necessary to maintain synaptic and behavioral states altered earlier in development. In contrast, overexpression of NL1FL in the fully mature adult was able to impair normal learning behavior after 1 month of expression. These results provide the first evidence that NL1 significantly impacts key developmental processes that permanently shape circuit function and behavior, as well as the function of fully developed neural circuits. Overall, these manipulations of NL1 function illuminate the significance of NL1 intracellular signaling in vivo, and enhance our understanding of the factors that gate the maturation of glutamatergic synapses and complex behavior. This has significant implications for our ability to address disorders such as autism spectrum disorders.