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Adaptive Regulatory Mechanisms of the Proteome and Phosphorylation Modifications in Multiple Organs of Mice during Torpor

ABSTRACT: Torpor is a crucial adaptive strategy employed by mammals to cope with low-temperature environments and/or seasonal food shortages. Investigating the torpor behavior of animals not only helps to elucidate their ecological adaptation mechanisms under conditions of food scarcity and harsh environments but also provides potential scientific insights for exploring organ transplantation, delaying aging, and extending lifespan. In this study, we established a torpor model using C57BL mice subjected to fasting and water deprivation at environmental temperatures of 22℃, 17℃, and 12℃. Through a combination of ultrastructural histology, proteomics, molecular biology, network analysis, and bioinformatics, we systematically analyzed the changes in protein expression and phosphorylation modification regulation in the kidney, liver, heart, and brown adipose tissue (BAT) of torpid mice. Notably, proteins such as phosphoenolpyruvate carboxykinase (PCK), tyrosine aminotransferase (TAT), and microtubule-associated protein 2 (MAP2) play significant roles during the torpor state in mice. These findings lay a crucial foundation for a deeper understanding of the molecular regulatory mechanisms underlying mammalian torpor and their tolerance strategies in adverse environments.

ORGANISM(S): Mus Musculus

SUBMITTER: Hui Wang

PROVIDER: PXD062891 | iProX | Fri Apr 11 00:00:00 BST 2025

REPOSITORIES: iProX

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Project description:Moderate caloric restriction (CR) and weight loss are beneficial for the promotion of health; however, there is controversy regarding the effects of dieting regimens on behavior. In this study, we investigated two different dieting regimens: repeated fasting and refeeding (RFR) and daily feeding of half the amount of food consumed by RFR mice (CR). Mice in both regimens were subjected to 20% reduction in food intake and transiently reduced their body weights during the first 12 days of the study. Open field, light-dark transition, elevated plus maze, and forced swimming tests indicated that CR, but not RFR, reduced anxiety- and depressive-like behaviors, with a peak on day 8. Using a mouse whole genome microarray, we analyzed gene expression in the prefrontal cortex, amygdala, and hypothalamus. In addition to the caloric restriction-responsive genes commonly modified by RFR and CR, each regimen differentially changed the expression of distinct genes in each region. The most profound change was observed in the amygdala of CR mice: 884 genes were specifically up-regulated. Ingenuity pathway analysis showed that these 884 genes significantly modified 9 canonical pathways in the amygdala. alpha-adrenergic and dopamine receptor signaling were the two top-scoring pathways. Quantitative real-time RT-PCR confirmed the up-regulation of 6 genes in these pathways. Ppp1r1b encoded Darpp-32 including dopamine receptor signaling, and the increased protein was specific for CR mice. Our results suggest that moderate CR may modify anxiety- and depressive-like behaviors and alter gene expression especially in the mouse amygdala. Experiment Overall Design: We tested three feeding regimens. One group of mice had ad libitum (AD) access to food, and was used as a control group. The repeated fasting and refeeding (RFR) group of mice were fasted on day 0 and allowed to feed ad libitum on day 1. Amounts of food consumed by the three mice on day 1 were measured. The half amount of food consumed by the RFR mice was given to three mice in another cage for two days (days 0 and 1) (CR group). The fasting followed by refeeding (RFR) and the restriction of food (CR) were repeated up to day 16. Body weights and consumed chow were measured at 20:00 every day, and then the fasting or feeding period was started. In RFR mice, fasting was started on day 0.Total RNA was prepared from the prefrontal cortex, hypothalamus, and amygdala in the mice of each group on day 8. An equal amount of RNA from 4 mice in each group was pooled and used for microarray analysis.

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Adaptive Regulatory Mechanisms of the Proteome and Phosphorylation Modifications in Multiple Organs of Mice during Torpor

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