ARRA: PLP Alternative splicing and oligodendrocyte differentiation

PLP and DM20 are major myelin proteins, generated from the same gene by alternative splicing of exon 3B. lAs PLP becomes the predominant isoform, the PLP/DM20 ratio increases in differentiated oligodendrocytes (OL) versus progenitors (OPC). Mutations, which impair the PLP alternative splicing and alter the PLP/DM20 ratio, are associated with Pelizaeus-Merzbacher disease and hereditary spastic paraplegias. The scope of the parent grant is to investigate the regulation of PLP/DM20 ratio in OL. We have made significant progress and have identified a synergistic regulation of the PLP/DM20 ratio by hnRNPH and F. We have identified additional hnRNPs and splicing factors, whose role in controlling PLP/DM20 ratio will be investigated in the supplement by using experimental design and methodologies proposed in the parent grant. In Aim 1, we will investigate the function of PSF and PTBP1 in controlling the PLP alternative splicing with knock down studies by RNAi and determine the role of PSF and PTB1 in the synergistic regulation of the PLPIDM20 ratio by hnRNPH and F. In Aim2, we will investigate the function of hnRNPA2/B1 and PUF60 and PUF60 variant 1, the expression of which increase in OL vs. OPC and correlates with the increase in PLP/DM20 ratio. We will investigate their function in controlling PLP/DM20 ratio with knock down studies by RNAi. Studies proposed in this supplement will create new jobs. retain currently employed personnel, benefit vendors through the purchase of equipment and molecular reagents, thus creating jobs in these sectors and significantly enhance the pace of discovery and research progress. The University of Kentucky, College of Medicine is a principal force for economic development in the state of Kentucky. Salary and benefits of more than 2,300 faculty, staff and learners are supported by research dollars generated by College of Medicine faculty. From these research efforts, more than $56 million is paid to these individuals in their quest for new knowledge. Additional extramural research funding is important for he University of Kentucky to maintain and grow its research portfolio as well as preserve and create jobs. If funded, this research program will create and/or retain 2 jobs.