Meeting Abstract
Temperature-responsive genes, such as those coding for heat shock proteins, play a vital role in embryogenesis. Cold-inducible RNA-binding protein (Cirp) is a heat shock protein present in the gonadal tissues of multiple taxa with a potential regulatory role in the sex-determining pathway. The red-eared slider turtle (Trachmeys scripta elegans ) exhibits temperature-dependent sex determination (TSD), where thermal cues trigger gonadal differentiation during development. In T. s. elegans, Cirp is up-regulated concurrent with estrogen-inducing transcripts at female-producing temperatures. Intron retention has been proposed as a regulatory mechanism for sex-specific development, and RNA-binding proteins can regulate the retention of introns. As Cirp is an RNA-binding protein localized in developing gonads, we hypothesize that Cirp plays a regulatory role in gonadogenesis by impacting the stability of target transcripts via intron retention. T. s. elegans eggs were incubated under fluctuating temperature treatments and either held under conditions that should produce males, or given a simulated heatwave to induce female development. Gonads from embryos were dissected for immunoprecipitation and RNA-seq. Sequenced RNA product will be aligned to an internal transcriptome developed from published raw reads to identify the target transcripts. Target transcripts will be translated and aligned to the painted turtle (Chrysemys picta) proteome to identify alignment gaps that correspond to retained introns. Our approach will help us understand how Cirp responds to fluctuating temperature treatments, and how it may regulate the nuclear expression of reproductive genes.
