Engineered synapse model cell: genetic construction and chemical evaluation for reproducible high-throughput analysis.
Bioassay models of neural functions must lend themselves to high-throughput analysis in neural drug discovery. However, smart analysis methods for these functions have not yet been fully established. Here, we describe the development of a synapse model for cell-based biosensing. The engineered synapse model cell expresses ionotropic glutamate receptor on its surface, like the neural postsynaptic membrane. The advantages of the model cell are the ease of handling and reproducibility as compared with the cultured neural cell, and it can be employed to evaluate receptor function through ion flux analysis. The agonist-induced sodium influx was monitored as an agonist concentration-dependent increase in the observed fluorescence signal. Furthermore, we found that our model cell enables the correction of uneven cellular signal levels using a reporter system. Our engineered synapse model cell can be employed as a powerful tool for the screening of lead substances in pharmaceutical high-throughput analysis.
Migita
Satoshi
S
Department of Biological Functions and Engineering, Kyushu Institute of Technology, Kitakyushu Science and Research Park, Fukuoka, 808-0196, Japan.
Tateishi
Akito
A
Keinänen
Kari
K
Haruyama
Tetsuya
T
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D019556:COS Cells; D002522:Chlorocebus aethiops; D055808:Drug Discovery; D005453:Fluorescence; D015870:Gene Expression; D018698:Glutamic Acid / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008959:Models, Neurological; D017470:Receptors, Glutamate / Q000235:genetics / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D013569:Synapses / Q000378:metabolism*; D014162:Transfection
D017470:Receptors, Glutamate; C109433:glutamate receptor type D; D049452:Green Fluorescent Proteins; D018698:Glutamic Acid; D012964:Sodium