Unknown,Transcriptomics,Genomics,Proteomics

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Deficient of IL-18 in mice leads to the reduction of neuropeptides in the amygdala related with behavioral change

ABSTRACT: Interleukin (IL)-18 is recognized as a multifunctional mediator in inflammation and immune responses. In addition, the production of IL-18 is stimulated by physiological and/or psychological stress implicated in several behavioral disorders. However, it still remains unclear about the role of IL-18 on the central nervous system. In this study, we analyzed gene expression of 6 brain regions; the olfactory bulb, prefrontal cortex, striatum, amygdala, hypothalamus, and hippocampus. Microarray analysis indicated the regional differences of expression in IL-18-/- mice. In particular, the most profound change was observed in the amygdala, in which 1,237 genes were differentially expressed. We purchased IL-18-/- mice and C57BL/6 mice from Charles River Japan . To produce F1 generation mice, three male IL-18-/- mice were mated with six female C57BL/6J mice. We mated F1 mice and created their F2 littermates. Six week-old F2 mice littermates were weaned. At 12 weeks, mice were sacrificed at 14:00 under general anesthesia with diethyl ether. After the systemic perfusion with cold phosphate-buffered saline through the heart using a syringe attached to a 21-G needle, the whole mouse brains were removed. Coronal brain sections (1 mm thick) were prepared on ice using a brain slicer . The olfactory bulb was sliced between 3.5 and 4.5 anterior to bregma. The prefrontal cortex was sliced between 2.5 and 3.5 mm anterior to bregma. The striatum was sliced between -0.5 and 0.5 mm posterior to bregma. The hypothalamus, the amygdala, and the hippocampus were sliced between 1.5 and 2.5 mm posterior to bregma. Total RNA was immediately prepared from these samples using TRIzol and purified by RNeasy kit.

ORGANISM(S): Mus musculus

SUBMITTER: Yuta Yamamoto

PROVIDER: E-GEOD-19522 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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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Deficient of IL-18 in mice leads to the reduction of neuropeptides in the amygdala related with behavioral change

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