Hypothalamic reprogramming by sperm microRNA

Nine microRNAs (miRs) previously identified as increased in the sperm of stressed sires were microinjected into C57/Bl6:129S6/SvEvTac mouse single cell zygotes to examine the hypothesis that specific sperm miRs function postfertilization to alter offspring stress responsivity. Derived mice were exmined for hypothalamic-pituitary-adrenal axis stress response in adulthood, and the paraventricular nucleus of the hypothalamus was subsequently collected for gene expression analysis by next gen sequencing. Similar to what we reported previously in our paternal stress model, the majority of diffferenitally expressed genes in the PVN exhibited decreased expression, supporting that an increase of specific sperm miRs in the zygote can elicit long-term genetic reprogramming. Futher, marked changes in the expression of extracellular matrix and collagen gene sets suggested altered blood-brain barrier permeability with potential consequences for neuroendocrine function. mRNA profiling of paraventricular nucleus from adult male mice derived from zygote microinjection of nine miRs (multi-miR; miR-29, miR-30a, miR-30c, miR-32, miR-193-5p, miR-204, miR-375, miR-532-3p, miR-698) or PBS. Six biological replicates per group (12 total cDNA libraries) were multiplexed and sequenced on two identical HiSeq2000 lanes (Illumina).