Myelination is essential for proper conduction across all neural networks. Myelination in the central nervous system is performed by glial cells called oligodendrocytes. Mature, myelinating glia develop from progenitor cells through distinct differentiation steps, and gene expression markers have been established that are unique to oligodendrocyte lineage cells. This study aims to establish a reliable RT-qPCR protocol to quantify oligodendrocyte-specific gene expression according to the MIQE guidelines. Primers specific to the progenitor, immature, and myelinating stages of oligodendrocyte differentiation were validated, showing 90-100% efficiency. Reference gene primers were also validated, and their stability was determined to make recommendations for those to use as normalization factors. The expression pattern of oligodendrocyte-specific genes across stages of postnatal development was similar to previously defined RNA-seq profiles in isolated cell types. The validated RT-qPCR assays developed build a framework for future investigation on myelination during development and remyelination in disease.
