A Head‐Specific Transcriptomic Study Reveals Key Regulatory Pathways for Winter Diapause in the Mosquito <i>Culex pipiens</i>

The primary vector of the West Nile virus, Culex pipiens, undergoes reproductive dormancy during the adverse winter season. While our current understanding has mainly focused on cellular signals and phenotypic shifts occurring at a global scale during diapause, information on tissue-specific transcriptomic changes remains limited. This knowledge gap is a major challenge in interpreting the regulatory mechanisms at the tissue level. To address this, the present work utilized RNA-seq technology to investigate the transcriptional changes in the head that house the brain and crucial endocrinal organs such as corpora allata. We obtained RNA samples from the heads of diapausing and nondiapausing female mosquitoes at two specific time intervals, ZT0 and ZT16, and then subjected them to sequencing. Our results revealed differences in differentially expressed genes between diapause and non-diapause at ZT0 and ZT16, highlighting the phenotypic and diel variations in gene expression. We also selected twelve genes associated with the diapause phenotype and examined the transcript abundance at six different time points over 24 h. qRT-PCR analysis showed similar up- and downregulation of transcripts between the diapause and nondiapause phenotypes thus validating the results of RNA-seq. In summary, our study identified new genes with phenotypic and diel differentiation in their expression, potentially linking photoperiod to seasonal reproductive dormancy in insects. The newly presented information will significantly advance our understanding of head-specific genes crucial for insect diapause.