Identification of neuron-specific promoters in Ciona intestinalis.
We isolated 5' flanking regions of four genes, Ci-Galphai1, Ci-arr, Ci-vAChTP, and Ci-vGAT, each of which is expressed in distinct sets of neurons in the central nervous system of the ascidian Ciona intestinalis, and we examined their function by introducing green fluorescent protein (GFP)-fusion constructs into Ciona embryos. The reporter gene driven by the 5' flanking region of Ci-Galphai1, Ci-arr, and Ci-vAChTP recapitulated the endogenous gene expression patterns, while that of Ci-vGAT can drive GFP expression in particular subsets of neurons expressing the endogenous gene. Deletion analysis revealed that the Ci-Galphai1 promoter consists of multiple regulatory modules controlling the expression in different types of cells. The GFP fluorescence enabled visualization of cell bodies and axons of different sets of neurons in ascidian larvae. These promoters can be a powerful tool for studying molecular mechanisms of neuronal development as well as neuron networks and functions in ascidians.
Yoshida
Reiko
R
Department of Life Science, Graduate School of Science, Himeji Institute of Technology, Hyogo 678-1297, Japan.
Sakurai
Daisuke
D
Horie
Takeo
T
Kawakami
Isao
I
Tsuda
Motoyuki
M
Kusakabe
Takehiro
T
D000595:Amino Acid Sequence; D000818:Animals; D002938:Ciona intestinalis / Q000196:embryology* / Q000235:genetics; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007814:Larva / Q000166:cytology; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D050495:Vesicular Inhibitory Amino Acid Transport Proteins
D044385:GTP-Binding Protein alpha Subunits; D006023:Glycoproteins; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; C493406:vesicular GABA transporter; D049452:Green Fluorescent Proteins