RNA sequencing analysis of PINCH mutant sciatic nerves

During peripheral nervous system (PNS) development, Schwann cells (SC) depend on interactions between extracellular matrix proteins, integrins and growth factor signal transduction pathways for survival, sorting and wrapping of axon segments to form myelin. Together with integrin-linked-kinase and parvins, PINCH (Particularly-interesting-new-cystidin-hystidin-rich protein) forms a heterotrimeric complex referred to as the IPP complex, which functions as a signaling hub connecting integrins with various signaling pathways. Using SC-specific conditional gene targeting in mice, gene expression profiling by RNAseq, and other methodologies, we demonstrate distinct functional roles for PINCH1 and PINCH2 in SC. Overall design: Total RNA was prepared from nerves collected from control, PINCH1 mutant and PINCH2 mutant (three biological replicates each). Each biological replicate consisted in a pool of four SN, from 1 male and 1 female to avoid gender-related differences). All procedures involving mice were in strict accordance to the guidelines approved by the Portuguese Veterinary Office (DGAV) and EU directives.