Physical activity as a promoter of stress resilience: An analysis of behavioral effects and brain connectivity with cytochrome c-oxidase activity in adult male Wistar rats.

Physical activity (PA) is very beneficial for physical and mental health. This study aims to examine the resilience-inducting effect of PA in adult male Wistar rats exposed to unpredictable chronic mild stress (UCMS). Furthermore, we analyzed the influence of PA on behavioral tasks and functional brain connectivity with cytochrome c oxidase technique. The cytochrome c oxidase (CCO) is a mitochondrial enzyme involved in oxidative phosphorylation and ATP generation. For this analysis, we included five groups: Basal (n = 10, to determine the basal level of brain activity), Behav (n = 15, subjected exclusively to behavioral tests), PA (n = 10, exposed to physical activity), UCMS (n = 15, subjected to a stress protocol) and PA + UCMS (n = 15, exposed to PA prior to stress). The UCMS protocol consisted of randomly presenting several different stressors over four consecutive weeks. We evaluated several behaviors of the Behav, UCMS, and PA + UCMS groups. This assessment includes the hedonic responses using the sucrose consumption task, unconditioned anxiety with the zero maze, and coping strategies assessed with the cat odor test. The UCMS group showed an anhedonia profile and increased anxiety compared with the other groups. Although in the exposure to cat odor test, the PA + UCMS remained for the same time in the cat odor compartment as the other groups, it did not approach the odor, showing that it detected the risk. This response is more adaptive than the responses of the UCMS and Behav groups. An exploratory analysis of the cerebral connections showed an increase in CCO activity in the UCMS group compared to the other groups. This overactivity was reduced in dorsal Cornu Ammonis 3(dCA3) by prior PA. In this region, PA + UCMS showed similar activity as the groups not subjected to chronic stress. Therefore, PA can prevent the harmful effects of chronic stress on dCA3.

An evaluation of hedonic responses in taste-potentiated odor aversion using the analysis of licking microstructure and orofacial reactivity.

Two experiments examined the hedonic responses conditioned to odor cues in the phenomenon of taste-potentiated odor aversion. Experiment 1 analyzed the microstructure of licking behavior during voluntary consumption. A tasteless odor (amyl acetate) was delivered to rats either diluted in water or mixed with saccharin before being injected with LiCl. At test, subjects which had received the odor-taste compound during conditioning showed both lower odor consumption and lick cluster size, a result indicating an increased negative evaluation of the odor. Experiment 2 examined the orofacial reactions elicited by the odor as index of its hedonic impact. During conditioning, the rats were intraorally infused with either the odor alone or the odor-saccharin compound before being injected with LiCl. At test, they were infused with the odor and their orofacial responses video recorded. More aversive orofacial responses were elicited by the odor cue in rats that had compound conditioning, again a result indicating a strengthened negative hedonic reactivity compared to animals experiencing odor aversion conditioning alone. Taken together, these results indicate that taste-mediated potentiation of odor aversion conditioning impacts on the acquisition of conditioned hedonic reactions as well as consumption.

Impact of environmental enrichment on the GABAergic neurons and glucocorticoid receptors in the hippocampus and nucleus accumbens of Wistar rats: Pro-resilient effects.

The unpredictable chronic mild stress (UCMS) model has been used to induce depressive-like symptoms in animal models. Our work aims to evaluate the impact of environmental enrichment on male Wistar rats in an animal model for depression. For this purpose, we aim to assess changes in GR and GABAergic (PV+) density in cerebral regions related to cognitive-affective processes associated with depressive disorder, such as the dorsal- ventral hippocampus and accumbens nuclei. Three groups of rats were used: UCMs (unpredictable chronic mild stress), EE+ UCMs (enrichment + stress) and CONT (behavioral tests only). Hedonic responses elicited by sucrose solution were examined by licking behavior analysis; the anxiety level was evaluated using the elevated zero maze and the forced swimming (passive coping) tests. The environmental enrichment reduced the effects of chronic stress, promoting greater resilience. Thus, the UCMs group showed an anhedonia response, more anxiety and immobility behavior than either the control or the EE+ UCMs groups. Regarding immunochemistry results, there was a reduction in GABAergic activity coupled with increased activation of GR in UCMs in the dorsal hippocampus, but there were no differences between groups in the ventral hippocampus. These results suggest environmental enrichment could enhance greater resilience, reducing the vulnerability of the subjects to develop disorders such as depression and anxiety.

Odor-taste pairings lead to the acquisition of negative hedonic qualities by the odor in aversion learning.

Three experiments examined the affective responses conditioned to an odorous stimulus in the taste-mediated odor aversion learning paradigm. Experiment 1 analyzed the microstructure of licking behavior during voluntary consumption. Before conditioning, water-deprived rats had access to a bottle containing either a tasteless odor (0.01% amyl acetate) diluted in water or mixed with 0.05% saccharin. Next, the rats were injected with either LiCl or saline immediately after drinking saccharin. At test, they received the odor and taste solutions on separate days. Lick cluster size was used as a direct measure of the hedonic response to the odor cue. Rats receiving odor-taste pairings prior to the saccharin devaluation showed both lower consumption and lick cluster size, reflecting a reduced hedonic evaluation of the odor. Experiments 2a and 2b used the orofacial reactivity method. After pretraining in the drinking boxes with the odor alone or mixed with saccharin, the rats were intraorally infused with saccharin before injection with LiCl or saline. At test, they were infused in separate sessions with the odor and taste and their orofacial reactions video recorded. There were increased aversive orofacial responses to the odor in rats that had prior odor-taste experience, a result indicating a negative hedonic evaluation of the odor. These results provide evidence of conditioned changes in affective value of odor cues through taste-mediated learning and are consistent with the idea that odor-taste pairings lead to the acquisition of taste qualities by the odor.

Reorganization of Brain Networks as a Substrate of Resilience: An Analysis of Cytochrome c Oxidase Activity in Rats.

The unpredictable chronic mild stress (UCMS) model has been used to induce depressive-like symptoms in animal models, showing adequate predictive validity. Our work aims to evaluate the effects of environmental enrichment (EE) on resilience in this experimental model of depression. We also aim to assess changes in brain connectivity using cytochrome c oxidase histochemistry in cerebral regions related to cognitive-affective processes associated with depressive disorder: dorsal hippocampus, prefrontal cortex, amygdala, accumbens, and habenula nuclei. Five groups of rats were used: UCMS, EE, EE + UCMS (enrichment + stress), BG (basal level of brain activity), and CONT (behavioral tests only). We assessed the hedonic responses elicited by sucrose solution using a consumption test; the anxiety level was evaluated using the elevated zero maze test, and the unconditioned fear responses were assessed by the cat odor test. The behavioral results showed that the UCMS protocol induces elevated anhedonia and anxiety. But these responses are attenuated previous exposure to EE. Regarding brain activity, the UCMS group showed greater activity in the habenula compared to the EE + UCMS group. EE induced a functional reorganization of brain activity. The EE + UCMS and UCMS groups showed different patterns of connections between brain regions. Our results showed that EE favors greater resilience and could reduce vulnerability to disorders such as depression and anxiety, modifying metabolic brain activity.

Characterizing Hedonic Responses to Flavors Paired with Internal Pain and Nausea through the Taste Reactivity Test in Rats.

Feeding behavior is a complex experience that involves not only sensory (i.e., visual, odor, taste, or texture) but also affective or emotional aspects (i.e., pleasure, palatability, or hedonic value) of foods. As such, behavioral tests that assess the hedonic impact of foods are necessary to fully understand the factors involved in ingestive behavior. In this protocol, we use the taste reactivity (TR) test to characterize the hedonic responses of rats to flavors paired with either lithium chloride-induced nausea or internal pain produced by hypertonic NaCl, two treatments that reduce voluntary consumption. This application of the TR test demonstrates how emetic and non-emetic (somatic pain in particular) treatments produce dissociable patterns of hedonic reactions to fluids: only emetic treatments result in the production of aversive orofacial responses, reflecting conditioned nausea, whereas somatic pain produces immobility, reflecting conditioned fear. Other methods, such as the microstructural analysis of licking behavior, do not reliably distinguish conditioned nausea and fear, a key advantage of the more selective TR procedure. This protocol also contains guidance for adaptation to other species and designs.