Recent studies in our laboratory have shown that the 32 kb myelin basic protein (MBP) transcription unit is contained within an overlapping transcription unit, termed the Golli-mbp gene, which is approximately 105 kb in the mouse. The Golli-mbp transcription unit consists of eleven exons of which parts or all of the last seven exons form the MBP transcription unit. Expression of the MBP transcription unit begins at a promoter upstream of exon 5b of the Golli-mbp gene, and expression of the Golli-mbp transcription unit begins at exon 1, producing a number of tissue, cell and developmentally regulated splice products at least three of which contain one or more exons found within the MBP transcription unit. The overall objective of this proposal is to determine the developmental and tissue-specific regulatory element of the Golli-mbp transcription unit in the mouse using in vitro transfection of cells and transgenic mice. We will analyze the role of these elements in modulating the expression of the MBP transcription with respect to levels of transcription as well as the tissue, cell and developmental expression of the gene. Initially, we will analyze the region upstream of Golli-mbp exon 1 for elements involved in the tissue, cell and developmental regulation of this transcription unit. We w ill also examine the possibility that one or more of these regulatory elements are found elsewhere in the gene, particularly the first intron. We will prepare constructs of putative regulatory regions plus a reporter gene and test them by transfection analysis in cell lines that do and do not express the Golli-mbp gene. We will then test the most promising constructs in transgenic animals to provide proof that the putative regulatory regions are operative in vivo. In other experiments we will examine the influence of the Golli-mbp promoter and any other regulatory elements we find on the levels and nature of expression of the MBP transcription unit. The deletion in the shiverer mouse genome affects at least two products of the Golli-mbp transcription unit in addition to MBPs. In view of the pleiotropic nature of the shiverer deletion on oligodendrocyte function we will introduce a Golli-mbp minigene into shiverer mice and carefully examine all the effects known to be associated with the shiverer mutation (beyond MBP expression and the """"""""shivering"""""""" phenotype). These studies could help us define the consequences of the expression of the Golli-mbp gene products. They would also help us determine if the produces themselves had an effect on the expression of the MBP transcription unit during oligodendrocyte development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS033091-01
Application #
2271658
Study Section
Neurology C Study Section (NEUC)
Project Start
1994-07-01
Project End
1998-06-30
Budget Start
1994-07-01
Budget End
1995-06-30
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Smith, Graham S T; Paez, Pablo M; Spreuer, Vilma et al. (2011) Classical 18.5-and 21.5-kDa isoforms of myelin basic protein inhibit calcium influx into oligodendroglial cells, in contrast to golli isoforms. J Neurosci Res 89:467-80
Paez, Pablo M; Fulton, Daniel J; Spreur, Vilma et al. (2010) Multiple kinase pathways regulate voltage-dependent Ca2+ influx and migration in oligodendrocyte precursor cells. J Neurosci 30:6422-33
Fisher, Robin; Xie, Yuan-Yun (2010) Growth defects in the dorsal pallium after genetically targeted ablation of principal preplate neurons and neuroblasts: a morphometric analysis. ASN Neuro 2:e00046
Xie, Yuan-Yun; Jacobs, Erin; Fisher, Robin (2009) Targeted ablation and reorganization of the principal preplate neurons and their neuroblasts identified by golli promoter transgene expression in the neocortex of mice. ASN Neuro 1:
Paez, Pablo M; Fulton, Daniel J; Spreuer, Vilma et al. (2009) Golli myelin basic proteins regulate oligodendroglial progenitor cell migration through voltage-gated Ca2+ influx. J Neurosci 29:6663-76
Paez, Pablo M; Spreuer, Vilma; Handley, Vance et al. (2007) Increased expression of golli myelin basic proteins enhances calcium influx into oligodendroglial cells. J Neurosci 27:12690-9
Jacobs, Erin C; Campagnoni, Celia; Kampf, Kathy et al. (2007) Visualization of corticofugal projections during early cortical development in a tau-GFP-transgenic mouse. Eur J Neurosci 25:17-30
Feng, Ji-Ming; Hu, Yanhong K; Xie, Lai-Hua et al. (2006) Golli protein negatively regulates store depletion-induced calcium influx in T cells. Immunity 24:717-27
Fernandes, Augustine O; Campagnoni, Celia W; Kampf, Kathy et al. (2004) Identification of a protein that interacts with the golli-myelin basic protein and with nuclear LIM interactor in the nervous system. J Neurosci Res 75:461-71
Feng, Ji-Ming; Fernandes, Augustine O; Campagnoni, Celia W et al. (2004) The golli-myelin basic protein negatively regulates signal transduction in T lymphocytes. J Neuroimmunol 152:57-66

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