Abstract

This project involves the application of recombinant DNA and immunochemical technologies to isolate and study cDNA clones of mRNAs specifically important to particular regions of the central nervous system. Using improved substractive hybridization procedures, we have isolated more than 150 cDNA clones of mRNAs expressed in cerebral cortex but not detectable in cerebellum; some of these clones have been partially characterized. Because most of these mRNAs are as abundant in hippocampus and caudate nucleus as in cerebral cortex, are present at much lower levels in diencephalon structures and are not detectable in either brain stem or cerebellum, the concept of a telencephalon-enriched battery of genes has emerged. The complete structures of the mRNAs showing the most interesting distributions will be determined by DNA sequence analysis and the encoded proteins will be studied for relationships to proteins whose sequences are known. Immunological reagents will be generated to identify and localize the protein in tissue extracts and in thin sections. To acquire functional information, we will use the """"""""anonymous"""""""" genes identified in this study to make new mutant mice affected in cortical function. Variants of the mouse analogues of these genes with a selectable marker inserted within their protein coding regions will be introduced by transformation into embryonic stem cells. Transformants will be screened for site specific integration (homologous recombination) by PCR. Embryonic stem cells heterozygous for such putative null mutations will be reintroduced into mice and germ line recipients will be bred to homozygosity. The phenotypes of mice homozygous for the disrupted versions of the genes will be analyzed to give an indication of the genes' function. As pilot studies, the genes for somatostatin and CCK will be disrupted. In related studies, the cellular phenotype of a mouse already identified to be deleted for one of the neuron- specific genes we have characterized by not grossly neurologically defective will be determined.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS022111-04A1
Application #
3404106
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1985-04-01
Project End
1991-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
4
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Scripps Research Institute
Department
Type
DUNS #
City
San Diego
State
CA
Country
United States
Zip Code
92037

  Publications

Iniguez, M A; De Lecea, L; Guadano-Ferraz, A et al. (1996) Cell-specific effects of thyroid hormone on RC3/neurogranin expression in rat brain. Endocrinology 137:1032-41
Chowdhury, D; Travis, G H; Sutcliffe, J G et al. (1995) Synaptotagmin I and 1B4 are identical: implications for synaptotagmin distribution in the primate brain. Neurosci Lett 190:9-12
Gerendasy, D D; Herron, S R; Jennings, P A et al. (1995) Calmodulin stabilizes an amphiphilic alpha-helix within RC3/neurogranin and GAP-43/neuromodulin only when Ca2+ is absent. J Biol Chem 270:6741-50
Ma, J; Norton, J C; Allen, A C et al. (1995) Retinal degeneration slow (rds) in mouse results from simple insertion of a t haplotype-specific element into protein-coding exon II. Genomics 28:212-9
Watson, J B; Coulter 2nd, P M; Margulies, J E et al. (1994) G-protein gamma 7 subunit is selectively expressed in medium-sized neurons and dendrites of the rat neostriatum. J Neurosci Res 39:108-16
Danielson, P E; Forss-Petter, S; Battenberg, E L et al. (1994) Four structurally distinct neuron-specific olfactomedin-related glycoproteins produced by differential promoter utilization and alternative mRNA splicing from a single gene. J Neurosci Res 38:468-78
Usui, H; Falk, J D; Dopazo, A et al. (1994) Isolation of clones of rat striatum-specific mRNAs by directional tag PCR subtraction. J Neurosci 14:4915-26
Godbout, M; Erlander, M G; Hasel, K W et al. (1994) 1G5: a calmodulin-binding, vesicle-associated, protein kinase-like protein enriched in forebrain neurites. J Neurosci 14:1-13
Danielson, P E; Watson, J B; Gerendasy, D D et al. (1994) Chromosomal mapping of mouse genes expressed selectively within the central nervous system. Genomics 19:454-61
Iniguez, M A; Morte, B; Rodriguez-Pena, A et al. (1994) Characterization of the promoter region and flanking sequences of the neuron-specific gene RC3 (neurogranin). Brain Res Mol Brain Res 27:205-14

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