We would like to understand how gene expression is controlled in cells of the nervous system and how that control leads to the formation of neural cell networks which can process information. A simple model of gene expression in nervous tissue will be established using the cloned gene for the mouse thy-1 glycoprotein, which we have recently isolated and characterized. This glycoprotein is the major cell-surface protein in adult brain and is evolutionarily related to the immunoglobulins, transplantation antigens and lymphoid differentiation antigens, molecules which play a crucial role in cellular recognition. We propose to characterize both alleles of the thy-1 gene from the mouse (thy-1.1 and thy-1.2) and construct a series of plasmid expression vectors containing this gene and dominant selectable markers. These genes will then be introduced into cell lines derived from mouse nerve and glial cells by DNA-mediated gene transfer. After transfection and stable integration, the expression of the cloned gene procuct on the cell surface will be assessed using allele-specific monoclonal antibodies. The levels of cell-surface expression will be compared to levels of expression of the endogenous gene in an attempt to identify critical regulators of gene expression. The DNA sequences required for thy-1 gene expression will be localized in modifying the cloned gene in vitro. These critical sequences can be compared with sequences necessary for the expression of this gene in lymphoid cells or in fibroblasts. The relative glycosylation pattern of the cloned gene produce can be compared to that of the endogenous thy-1 gene in lymphoid and neural cells. Finally, to assess possible functions for this gene product in cellular interactions in the nervous system, specific modifications will be made in the thy-1 protein by manipulating the gene in vitro. The reconstitution of gene expression at physiological levels using a cloned gene in an expression vector may allow (1) identification of regulators of other neural-specific genes, (2) identification of gene families which are expressed during neuron differentiation, or (3) development of techniques to allow nerve cell function to be specifically altered. The long-term goals of this project are to understand how gene expression during development controls the interaction of neural cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033868-02
Application #
3284002
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1984-08-01
Project End
1987-07-31
Budget Start
1985-08-01
Budget End
1986-07-31
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Salk Institute for Biological Studies
Department
Type
DUNS #
005436803
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Selleri, L; Giovannini, M; Romo, A et al. (1994) Cloning of the entire FLI1 gene, disrupted by the Ewing's sarcoma translocation breakpoint on 11q24, in a yeast artificial chromosome. Cytogenet Cell Genet 67:129-36
Saleh, M; Selleri, L; Evans, G A (1993) A novel zinc finger gene on human chromosome 1qter that is alternatively spliced in human tissues and cell lines. Am J Hum Genet 52:192-203
Sun, W; McPherson, J D; Hoang, D Q et al. (1992) Mapping of a human brain voltage-gated calcium channel to human chromosome 12p13-pter. Genomics 14:1092-4
Giovannini, M; Selleri, L; Biegel, J A et al. (1992) Interphase cytogenetics for the detection of the t(11;22)(q24;q12) in small round cell tumors. J Clin Invest 90:1911-8
Eubanks, J H; Djabali, M; Selleri, L et al. (1992) Structure and linkage of the D2 dopamine receptor and neural cell adhesion molecule genes on human chromosome 11q23. Genomics 14:1010-8
Sun, W; Ferrer-Montiel, A V; Schinder, A F et al. (1992) Molecular cloning, chromosomal mapping, and functional expression of human brain glutamate receptors. Proc Natl Acad Sci U S A 89:1443-7
Eubanks, J H; Altherr, M; Wagner-McPherson, C et al. (1992) Localization of the D5 dopamine receptor gene to human chromosome 4p15.1-p15.3, centromeric to the Huntington's disease locus. Genomics 12:510-6
Saleh, M; Selleri, L; Little, P F et al. (1992) Isolation and expression of linked zinc finger gene clusters on human chromosome 11q. Genomics 14:970-8
Selleri, L; Eubanks, J H; Giovannini, M et al. (1992) Detection and characterization of ""chimeric"" yeast artificial chromosome clones by fluorescent in situ suppression hybridization. Genomics 14:536-41
Ahmed, C M; Ware, D H; Lee, S C et al. (1992) Primary structure, chromosomal localization, and functional expression of a voltage-gated sodium channel from human brain. Proc Natl Acad Sci U S A 89:8220-4

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