Abstract
External cues that dictate the direction of cell migration are likely dynamic during many biological processes such as embryonic development and wound healing. Until recently, how cells integrate spatial and temporal information to determine the direction of migration has remained elusive. In Dictyostelium discoideum, the chemoattractant cAMP that directs cell aggregation propagates as periodic waves. In light of the fact that any temporally evolving complex signals, in principle, can be expressed as a sum of sinusoidal functions with various frequencies, the Dictyostelium system serves as a minimal example, where the dynamic signal is in the simplest form of near sinusoidal wave with one dominant frequency. Here, we describe a method to emulate the traveling waves in a fluidics device. The text provides step-by-step instructions on the device setup and describes ways to analyze the acquired data. These include quantification of membrane translocation of fluorescently labeled proteins in individual Dictyostelium cells and estimation of exogenous cAMP profiles. The described approach has already helped decipher spatial and temporal aspects of chemotactic sensing in Dictyostelium. More specifically, it allowed one to discriminate the temporal and the spatial sensing aspects of directional sensing. With some modifications, one should be able to implement similar analysis in other cell types.
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Lauffenburger D, Farrell B, Tranquillo R et al (1987) Gradient perception by neutrophil leucocytes, continued. J Cell Sci 88:415–416
Futrelle RP (1982) Dictyostelium chemotactic response to spatial and temporal gradients. Theories of the limits of chemotactic sensitivity and of pseudochemotaxis. J Cell Biochem 18:197–212
Xu X, Meier-Schellersheim M, Yan J, Jin T (2007) Locally controlled inhibitory mechanisms are involved in eukaryotic GPCR-mediated chemosensing. J Cell Biol 178:141–153
Zhang S, Charest PG, Firtel RA (2008) Spatiotemporal regulation of Ras activity provides directional sensing. Curr Biol 18:1587–1593
Fisher PR, Merkl R, Gerisch G (1989) Quantitative analysis of cell motility and chemotaxis in Dictyostelium discoideum by using an image processing system and a novel chemotaxis chamber providing stationary chemical gradients. J Cell Biol 108:973–984
Vicker MG (1994) The regulation of chemotaxis and chemokinesis in Dictyostelium amoebae by temporal signals and spatial gradients of cyclic AMP. J Cell Sci 107:659–667
van Haastert PJ (1983) Sensory adaptation of Dictyostelium discoideum cells to chemotactic signals. J Cell Biol 96:1559–1565
Tani T, Naitoh Y (1999) Chemotactic responses of Dictyostelium discoideum amoebae to a cyclic AMP concentration gradient: evidence to support a spatial mechanism for sensing cyclic AMP. J Exp Biol 202:1–12
Ebrahimzadeh PR, Högfors C, Braide M (2000) Neutrophil chemotaxis in moving gradients of fMLP. J Leukoc Biol 67:651–661
Irimia D, Liu SY, Tharp WG et al (2006) Microfluidic system for measuring neutrophil migratory responses to fast switches of chemical gradients. Lab Chip 6:191–198
Meier B, Zielinski A, Weber C et al (2011) Chemotactic cell trapping in controlled alternating gradient fields. Proc Natl Acad Sci U S A 108:11417–11422
Nakajima A, Ishihara S, Imoto D, Sawai S (2014) Rectified directional sensing in long-range cell migration. Nat Commun 5:5367
Acknowledgements
This work was supported by grants from the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research on Innovative Areas (23111506, 25111704) (to S.S.), JSPS Grant-in-Aid for Young Scientists (A) (22680024, 25710022) (to S.S.), JSPS Grant-in-Aid for Young Scientists (Start-up) and (B) (23870006, 25840069) (to A.N.), Japan Science and Technology Agency (JST) Precursory Research for Embryonic Science and Technology (PRESTO) (to S.S.), Platform for Dynamic Approaches to Living System from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and in part by the Human Frontier Science Programme (RGY 70/2008) and JSPS Grant-in-Aid for Scientific Research on Innovative Areas (25103008) (to S.S.).
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Nakajima, A., Sawai, S. (2016). Dissecting Spatial and Temporal Sensing in Dictyostelium Chemotaxis Using a Wave Gradient Generator. In: Jin, T., Hereld, D. (eds) Chemotaxis. Methods in Molecular Biology, vol 1407. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3480-5_8
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DOI: https://doi.org/10.1007/978-1-4939-3480-5_8
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