The long-term goals of this project are to elucidate the function of a G-rich 19-bp sequence in intron 3 of the PLP gene in PLP-DM20 splice site selection in vivo. The investigators have shown that deletion of this sequence causes a neurological disease in humans and that this sequence regulates selection of PLP splicing in oligodendrocytes in vitro. G-rich elements have been shown to regulate splice site selection in chimeric genes in vitro. The investigators propose to generate a line of knock-in mice carrying deletion of the G-rich 19-bp sequence and to utilize these mice to investigate PLP specific splicing in brain and peripheral nerves during development. In these studies, the investigators will characterize splicing at the early stages of myelination, peak of myelination, and adulthood. The production of PLP and other myelin proteins and myelin formation will be studied. The mouse phenotype will be characterized by motor testing. To the investigators' knowledge, mouse models designed to investigate in vivo regulation of alternative splicing are not available and none of the PLP mouse mutants allows studies of PLP splicing. The mouse model to be made will allow investigating the regulation of alternative splicing in the PLP gene in the context of the entire gene and in the presence of external cues. In addition, it has the potential to generate data of broader biological relevance about splicing in many other genes.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Small Research Grants (R03)
Project #
7R03HD043104-03
Application #
6897372
Study Section
Pediatrics Subcommittee (CHHD)
Program Officer
Oster-Granite, Mary Lou
Project Start
2004-05-25
Project End
2007-02-28
Budget Start
2004-05-25
Budget End
2007-02-28
Support Year
3
Fiscal Year
2004
Total Cost
$73,649
Indirect Cost
Name
University of Kentucky
Department
Neurology
Type
Schools of Medicine
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Wang, Erming; Dimova, Neviana; Sperle, Karen et al. (2008) Deletion of a splicing enhancer disrupts PLP1/DM20 ratio and myelin stability. Exp Neurol 214:322-30
Bohmer, Ralph M; Stroh, Helene P; Johnson, Kirby L et al. (2002) Fetal cell isolation from maternal blood cultures by flow cytometric hemoglobin profiles. Results of a preliminary clinical trial. Fetal Diagn Ther 17:83-9