Abstract
Embryonic Dissociated Dorsal Root Ganglia cultures are often used to investigate the role of novel molecular pathways or drugs in Schwann cell development and myelination. These cultures largely recapitulate the order of cellular and molecular events that occur in Schwann cells of embryonic nerves. However, the timing of Schwann cell developmental transitions, notably the transition from Schwann Cell Precursors to immature Schwann cells, has not been estimated so far in this culture system. In this study, we use RTqPCR to determine the expression profiles of Schwann cell developmental genes during the first week of culture. We first identified stable reference genes that show minimal variation across different experimental time points. Consequently, we normalized the mRNA profiles of Schwann cell developmental genes using the best internal reference genes. We then compared our data to the expression profiles of these genes in developing spinal nerves elaborated in numerous high-throughput and lineage tracing studies. This comparison helped in identifying that Schwann Cell Precursors transition into immature Schwann Cells between the 5th and 7th day in vitro. In effect, our data allows for a better understanding and interpretation of DRG culture experiments especially in studies that aim to elucidate the role of a novel gene in Schwann Cell development and myelination.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Funding: No specific funding was sought for this study. VKS is funded by a PhD fellowship from the French Ministry of Research and Innovation and Université de Paris. RB is funded by a doctoral contract from INSERM U1016, Institut Cochin, Paris.