MEASURING GLUTAMATE AND OXYGEN IN BRAIN REWARD CIRCUITS IN ANIMAL MODELS OF COCAINE ABUSE AND DECISION-MAKING

Drug-specific reward and associated effects on neural signaling are often studied between subjects, where one group self-administers drug and a separate group self-administers a natural reinforcer. However, exposure to drugs of abuse can cause long-term neural adaptations that can affect how an organism responds to drug reward, natural reward, and their reward-associated stimuli. Thus, to isolate drug-specific effects it is important to use models that expose the same organism to all of the aforementioned. Multiple schedules provide a means of dissociating the rewarding effects of a drug from the rewarding effects of food within a single animal. Further, drug users do not take drugs in isolation; rather, they are often faced with several concurrently available commodities (e.g. monetary goods, social relationships). Thus, using choice measures to assess the relative subjective value of drug reinforcers in both humans and animals promotes a translational understanding of mechanisms that govern drug-associated decision-making. Thus, in order to gain a more translational view of the neurobehavioral mechanisms that underlie drug-associated behavior, in the first study, glutamate was measured in the nucleus accumbens core (NAcC) and prefrontal cortex (PrL) in freely-moving rats as they behaved in a cocaine-food multiple schedule procedure. In the second study, oxygen dynamics were measured in the orbitofrontal cortex (OFC) of freely-moving rats as they behaved in a cocaine/food choice procedure. The results from the first study showed that, in the NAc and PrL, there was an increase in glutamate release when animals earned cocaine. Further, the number of glutamate peaks that occurred per cocaine lever press and per cocaine reinforcer was increased compared to food. In the second study, OFC oxygen dynamics were positively correlated with cocaine/food choice and generally tracked preference. Further, OFC oxygen dynamics were greater to cocaine related events. Taken together, these results showed the feasibility of combining electrochemical measurements with complex drug-related behavioral procedures. These results also highlight the importance of the PrL, NAcC, and OFC in the valuation of drug and non-drug commodities. Overall, these results add to our understanding of the neurobehavioral mechanisms that guide drug-associated behavior and create more precise experimental avenues to research potential treatments

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field