| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
Choline acetyltransferase (ChAT), the biosynthetic enzyme for ACh, is the most specific marker for the cholinergic neuron. Using the molecular biological techniques, we previously produced an antiserum to rat ChAT and established the immunohistochemistry. Recently we found that the antiserum stains cholinergic neurons in paraffin-embedded sections, and this enables us to study the phylogeny and comparative anatomy of the cholinergic neuron. We found, hoever, that cross-species reactivity of the antiserum is not wide enough to complete the present project. We, therefore, produced an antiserum to vesicular ACh transporter (VAChT), another specific protein for the cholinergic neuron, and examined whether or not it can be used for the present project. For this, using antisear to and digoxigenin-laveled RNA proves for ChAT and VAChT,we compared extensively the distribution of mRNAs and the corresponding proteins for ChAT and VAChT by immunohistochemistry and in situ hybridization histochemistry, both alone and with various combinations of them, and demonstrated that the antiserum to VAChT can be used for the present project.
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