Abstract
GPCR signaling is the most prevailing molecular mechanism for detecting ambient signals in eukaryotes. Chemotactic cells use GPCR signaling to process chemical cues for directional migration over a broad concentration range and with high sensitivity. Dictyostelium discoideum is a classical model, in which the molecular mechanism underlying eukaryotic chemotaxis has been well studied. Here, we describe protocols to evaluate the spatiotemporal chemotactic responses of Dictyostelium discoideum by different microscopic observations combined with biochemical assays. First, two different chemotaxis assays are presented to measure the dynamic concentration ranges for different cell strains or chemotactic parameters. Next, live-cell imaging and biochemical assays are provided to detect the activities of GPCR and its partner heterotrimeric G proteins upon chemoattractant stimulation. Finally, a method for detecting how a cell deciphers chemical gradients is described.
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Acknowledgments
We would like to thank NBRP-slime mold for distributing the bioresources described in this chapter. This work was supported by grants from JSPS KAKENHI Grant Numbers 17K07396 and 20K06631 to Y.K, and 19H00982 to M.U. This research was supported in part by AMED-CREST from Japan Agency for Medical Research and Development, AMED JP20gm0910001 to M.U.
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Kamimura, Y., Ueda, M. (2021). GPCR Signaling Regulation in Dictyostelium Chemotaxis. In: Kim, SB. (eds) Live Cell Imaging. Methods in Molecular Biology, vol 2274. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1258-3_27
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DOI: https://doi.org/10.1007/978-1-0716-1258-3_27
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