Fig. 1.
Molecular cloning and expression pattern of 5-HTapAC1. (A) Multiple sequence alignments of TM5 and TM7 of invertebrate 5-HT7 receptors. (B) Deduced amino acid sequence of 5-HTapAC1. Seven transmembrane (TM) domains are indicated and numbered. Predicted phosphorylation sites for PKA (●) and PKC (*) are marked. Triangles (▴) indicate N-glycosylation sites. (C) Multiple tissue RT-PCR analysis. AG, abdominal ganglia; BG, buccal ganglia; BM, buccal mass; CG, central ganglia (cerebral, pleural and pedal); GL, gill; OVT, ovotestis. (D) Single cell RT-PCR analysis of 5-HTapAC1. 5-HTapAC1 was expressed in sensory neurons and LFS, L7 motor neurons. The sensory neuron specific gene sensorin was expressed only in sensory cells. The housekeeping gene S4 was used as a control. Total RNA isolated from the whole abdominal ganglion was used as another control. SN, sensory neuron; LSF, LSF motor neuron; L7, L7 motor neuron; Ab. g., total RNA from the abdominal ganglion. (E) In situ hybridization of 5-HTapAC1 mRNA in sensory clusters. Dorsal abdominal ganglia and left pleural ganglia were shown. 5-HTapAC1 was expressed in sensory cells in the sensory cluster in pleural ganglia, as well as in L7 motor neurons. LC, left pleuroabdominal connective; RC, right pleuroabdominal connective; PN, pericardial nerve; BN, branchial nerve; L7, L7 motor neuron; LPI1, left pleural giant neuron; SenC, sensory cluster; c-pl, cerebro-pleural connective; pl-p, pleuropedal connective. (Scale bars, 500 μm for Upper; 200 μm Lower.) (F) In situ hybridization of 5-HTapAC1 mRNA in sensory-to-motor coculture. (Right) Shown are the hybridization with sense nucleotide probe. (Scale bar, 50 μm.)