MicroRNA expression analysis of livers of F1 male offspring fathered from control mice or stressed mice

Psychological stress is highly prevalent in modern society. Both epidemiologic and experimental animal studies demonstrate that chronic psychological stress exerts adverse effects on the initiation and/or progression of many diseases. Here, using a mouse model of restraint stress, we report the discovery of a novel signaling pathway linking paternal stress exposure to the reprogram of hepatic gluconeogenesis in the offspring. Offspring fathered by stressed males (Stress-F1) exhibits hyperglycemia as a result of enhanced hepatic gluconeogenesis, conpared to offspring from control fathers (Control-F1). Mechanistically, protein levels of PEPCK, a key gluconeogenic enzyme, were significantly increased, while its mRNA levels were unaffected in the stress-F1 mice, pointing to a posttranscriptional regulatory mechanism. Because MicroRNAs often play an important role in regulating gene expression at the posttranscriptional level, we investigated the expression profile of MicroRNAs in livers from Control-F1 and Stress-F1 mice. Liver tissues were collected from 12-week-old male control-F1 and stress-F1 mice (n=3 for each group). After having passed RNA quantity measurement using the NanoDrop 1000, the samples were labeled using the miRCURY™ Hy3™/Hy5™ Power labeling kit and hybridized on the miRCURY™ LNA Array (v.18.0). Following the washing steps the slides were scanned using the Axon GenePix 4000B microarray scanner.