This application proposes studies directed towards the identification and characterization of novel brain specific proteins. Rat brain cDNA clones were selected which correspond to mRNAs expressed in brain but not in liver or kidney. The nucleotide sequence of one such clone (plB236) has been determined, providing the amino acid sequence of the protein (1B236) encoded by the cloned mRNA. Antibodies raised against synthetic peptides corresponding to regions of this sequence detect immunoreactivity in a series of widely distributed neuronal structures in rat brain distinct from any other immunocytochemically defined neuronal system. In addition, the amino acid sequence of 1B236 has structural features in common with neuropeptide precursors, suggesting that the 1B236 protein may be cleaved to generate several novel neuropeptides. Preliminary evidence indicates that 1B236 immunoreactivity can be detected in rat brain extracts as both a 100,000 dalton glycoprotein and as processed peptides (15-40 amino acids). Synthetic peptides corresponding to 1B236 sequences have behavioral and electrophysiological activity. It is the aim of the proposed studies to: 1. Determine the predominant molecular forms of 1B236 immunoreactivity in rat brain and their relative amounts. 2. Isolate and characterize the 1B236 protein from rat brain. 3. Determine the exact forms of any peptides generated from the 1B236 protein in vivo and released in vitro from nerve terminals. 4. Deduce the sequence of steps involved in the post-translational processing of the 1B236 protein. 5. Determine the structure of the gene coding for the 1B236 protein. 6. Investigate the expression of the 1B236 protein and its products during development of the rat. 7. Test for similar molecules in other species. These studies will generate structural, genetic and developmental information on a novel brain specific protein which may represent a previously undetected neurotransmitter system. The data will provide the basis for functional testing of this protein and its peptide products and may provide a new avenue for the elucidation of neurologic disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS020728-02
Application #
3401283
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1984-04-01
Project End
1987-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
San Diego
State
CA
Country
United States
Zip Code
92037
Milner, R J (1990) Differential cloning approaches to the nervous system. Ann N Y Acad Sci 579:273-80
Milner, R J; Bloom, F E; Sutcliffe, J G (1987) Brain-specific genes: strategies and issues. Curr Top Dev Biol 21:117-50
Lai, C; Brow, M A; Nave, K A et al. (1987) Two forms of 1B236/myelin-associated glycoprotein, a cell adhesion molecule for postnatal neural development, are produced by alternative splicing. Proc Natl Acad Sci U S A 84:4337-41
Lai, C; Watson, J B; Bloom, F E et al. (1987) Neural protein 1B236/myelin-associated glycoprotein (MAG) defines a subgroup of the immunoglobulin superfamily. Immunol Rev 100:129-51
Milner, R J; Randolph, L; Bahr, D et al. (1987) Molecular biological approaches to the brain and their application to the study of alcoholism. Prog Clin Biol Res 241:291-302
Miller, F D; Naus, C C; Higgins, G A et al. (1987) Developmentally regulated rat brain mRNAs: molecular and anatomical characterization. J Neurosci 7:2433-44
Lenoir, D; Battenberg, E; Kiel, M et al. (1986) The brain-specific gene 1B236 is expressed postnatally in the developing rat brain. J Neurosci 6:522-30
Milner, R J; Lai, C; Lenoir, D et al. (1986) Brain-specific gene expression. Biochem Soc Symp 52:107-17
Bloom, F E; Battenberg, E; Ferron, A et al. (1985) Neuropeptides: interactions and diversities. Recent Prog Horm Res 41:339-67
Bloom, F E; Battenberg, E L; Milner, R J et al. (1985) Immunocytochemical mapping of 1B236, a brain-specific neuronal polypeptide deduced from the sequence of a cloned mRNA. J Neurosci 5:1781-802

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