|Budget Amount *help
¥1,600,000 (Direct Cost : ¥1,600,000)
Fiscal Year 1992 : ¥500,000 (Direct Cost : ¥500,000)
Fiscal Year 1991 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1990 : ¥700,000 (Direct Cost : ¥700,000)
Three years ago, we started our project entitled, "Genetic analysis of cholinergic neurons". We established the method for the screening of mutants showing abnormalities in cholinergic neurons. With this method we isolated several mutants and identified genes contributing to the mutants. Of these genes, we focused our work on genes functioning at the presynaptic terminals.
Seven such genes (unc-11, unc-13, unc-17, unc-18, unc-41, unc-63 and unc-64) were identified. We found the genes function in the regulatory pathway controlling acetylcholine levels. To investigate further these genes at the molecular level, we cloned the unc-18 gene, one of the seven genes. DNAs of the genomic and the cDNA clone were sequenced and the deduced amino acid sequence of the cDNA was determined. Comparison of the predicted amino acid sequence revealed has a role in the release of the neurotransmitters.
We cloned the cDNA into the expression vector, which permit to produce the unc-18 gene product and made antisera against the gene product. With the antisera we stained animals and found the gene product is specifically localized at motor neurons. These findings indicate that UNC-18 has a role in the axonal transport and influences acetylcholine flow in motor neurons.