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Comments on Ishihara T et al. (1995) International C. elegans Meeting "CONSTRUCTION OF FLUORESCENT MARKERS SPECIFIC TO THE NERVOUS SYSTEM OF C. ELEGANS" (0)
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Ishihara T, & Katsura I (1995). CONSTRUCTION OF FLUORESCENT MARKERS SPECIFIC TO THE NERVOUS SYSTEM OF C. ELEGANS presented in International C. elegans Meeting. Unpublished information; cite only with author permission.
To elucidate how the neuronal circuit is constructed, it may be useful to devise an easy way to observe the morphology of the nervous system in living animals. We are developing GFP markers that are expressed in various sets of neurons in C. elegans. The neuron specific markers are prepared by "promoter trapping" and by using promoters of putatively neuronal genes as predicted by the C. elegans genome project. From about 25,000 clones of a genomic library for promoter trapping constructed in lacZ fusion vector, we isolated three positive clones, of which the lacZ gene was exchanged subsequently for the GFP cDNA. One of them gives GFP expression in almost all neurons, while another gives very specific expression in the AFD cells. The C. elegans genome project predicts some neuronal genes by sequence homology. Among those, promoters of a glutamate receptor gene and a neurotransmitter transporter gene lead to expression in some interneurons and in the dopaminergic neurons, respectively. To identify the marked neurons or to observe neurites of these neurons, we use GFP fused with a nuclear localization signal or with a part of the Unc-76 protein as a marker, respectively. Using these markers, we hope to identify mutations causing defects in neural differentiation or connectivity. This method should be applicable even to lethal mutations, which prevent behavioral assay to detect neuronal disorder.