Bulk RNA-Seq of male and female lumbosacral dorsal root ganglia after VEGFA-instillation
This study examined transcriptomics in lumbosacral dorsal root ganglia that carries bladder afferent signals in a bladder-origin visceral pain animal model. C57BL6/J male and female mice underwent intravesical instillations of VEGF165, a mouse recombinant of VEGFA to induce visceral hypersensitivity. Saline-instilled male and female mice were used as controls. Across the four experimental groups, we found significant sex differences in the transcriptome profile in both saline-instilled animals and VEGF-induced signaling pathway changes in the lumbosacral sensory ganglia. These findings provide insight into potential molecular targets for alleviating visceral pain symptoms when considering sex as a biological variable. Overall design: Total of 16 C57BL6/J mice were used in this study.  Four experimental groups are Saline-instilled male mice (n=4) Saline-instilled female mice (n=4),VEGFA -instilled male mice (n=4) and VEGFA -instilled female mice (n=4). Intravesical instillations of VEGF165 (6.41 nM in 100 µL saline) or saline (100µL) were instilled  on Day 1,Day 5 and Day 9.  Manual Von Frey testing (VFT) was used to assess visceral mechanical sensitivity on Day 16. Four animals from each group were euthanized and their lumbosacral L1, L2, L6, S1, and S2 DRGs were harvested immediately. Total RNA was extracted using a RNAeasy miniprep kit (Qiagen, CA, USA). Extracted RNA was resuspended in 15 µL of RNase-free water and stored at -80°C. QC passed 300 ng of RNA per sample was used for library preparation using  universal mRNA Poly-A tail prep kit.Samples were sequenced on  Illumina NovaSEQ 6000 platform  (Paired End 2x150bp) at a depth of 40 million reads per sample.