Prenatal Heat Calls Program DNA Methylation in the Embryonic Zebra Finch Brain

We used reduced representation bisulfite sequencing (RRBS) to profile DNA methylation in whole brains of embryonic day 14 zebra finch embryos exposed prenatally to either heat call playback or control call playback from embryonic day 10 through embryonic day 13.

The study included embryos from both treatment groups, and samples failing RRBS quality control were excluded before downstream analysis, resulting in a final cohort of 35 embryos. Differential methylation analysis identified CpG sites associated with prenatal heat call exposure in genes and pathways related to nuclear receptor signaling, protein phosphorylation, synaptic organization, and GABAergic signaling. A recurrent signal across analytical models was intronic hypermethylation in RBFOX2, a gene involved in RNA splicing and neuronal development.