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The Journal of Neuroscience, January 1, 2000, 20(1):206-218
Visualization of Cranial Motor Neurons in Live Transgenic Zebrafish Expressing Green Fluorescent Protein Under the Control of the Islet-1 Promoter/Enhancer
Shin-ichi Higashijima1, 2, Yoshiki Hotta3, and Hitoshi Okamoto4 1 Inheritance and Variation Group, Precursory Research for Embryonic Science and Technology, Japan Science and Technology Corporation, Honmachi, Kawaguchi, Saitama 332-0012, Japan, 2 Division of Morphogenesis, National Institute for Basic Biology, Myodaijicho, Okazaki, Aichi 444-8585, Japan, 3 National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan, and 4 Laboratory for Developmental Gene Regulation, Brain Science Institute, RIKEN (The Institute of Physical and Chemical Research), Hirosawa, Wako, Saitama 351-0198, Japan
We generated germ line-transmitting transgenic zebrafish that express green fluorescent protein (GFP) in the cranial motor neurons. This was accomplished by fusing GFP sequences to Islet-1 promoter/enhancer sequences that were sufficient for neural-specific expression. The expression of GFP by the motor neurons in the transgenic fish enabled visualization of the cell bodies, main axons, and the peripheral branches within the muscles. GFP-labeled motor neurons could be followed at high resolution for at least up to day four, when most larval neural circuits become functional, and larvae begin to swim and capture prey. Using this line, we analyzed axonal outgrowth by the cranial motor neurons. Furthermore, by selective application of DiI to specific GFP-positive nerve branches, we showed that the two clusters of trigeminal motor neurons in rhombomeres 2 and 3 innervate different peripheral targets. This finding suggests that the trigeminal motor neurons in the two clusters adopt distinct fates. In future experiments, this transgenic line of zebrafish will allow for a genetic analysis of cranial motor neuron development.
Key words: zebrafish; neuron-specific promoter; transgenic; Islet-1; motor neuron; GFP; live visualization
Copyright © 2000 Society for Neuroscience 0270-6474/0/201206-13$05.00/0
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