[Establishment of a cell based high throughput screening model for transient receptor potential V3 modulators].
We established a cell based high throughput screening model by calcium assay on fluorometric imaging plate reader for finding modulators of TRPV3. The TRPV3 expression vector was transfected into HEK-293 and stable cell line expressing TRPV3 was selected with antibiotics. Upon TRPV3 specific modulators stimulated, pharmacological characteristics of TRPV3 over expression cell line were detected by calcium assay on fluorometric imaging plate reader. Assay conditions were optimized and stability of the model was observed. The reliability and accuracy of application to 96 and 384 well format high throughput screening were also evaluated. A stable HEK-293 cell line highly expressing TRPV3 was established. TRPV3 specific modulators could modulate calcium signal through TRPV3 in dose dependent manner. Optimized screening condition was established by assay development. This model is stable and sensitive, and meets the requirement of high throughput screening by Z'factor validation and Spiking test. This cell model can be applied to screening TRPV3 modulators by calcium assay.
Wang
Shaopeng
S
Beijing Genomics Institute, Chinese Academy of Sciences, Beijing 101300, China. wangshp@genomics.org.cn
Li
Jixian
J
Peng
Zhigang
Z
Cao
Jing
J
D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D017136:Ion Transport; D007668:Kidney / Q000166:cytology; D049990:Membrane Transport Modulators / Q000378:metabolism*; D008954:Models, Biological; D050916:TRPV Cation Channels / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D049990:Membrane Transport Modulators; D050916:TRPV Cation Channels; C459445:TRPV3 protein, human; D002118:Calcium