Chemotaxis: Under Agarose Assay

Methods Mol Biol. 2022:2364:327-338. doi: 10.1007/978-1-0716-1661-1_16.

Abstract

The unicellular eukaryotic amoeba, Dictyostelium discoideum, represents a superb model for examining the molecular mechanism of chemotaxis. Under vegetative conditions, the amoebae are chemotactically responsive to pterins, such as folic acid. Under starved conditions, they lose their sensitivity to pterins and become chemotactically responsive to cAMP. As an NIH model system, Dictyostelium offers a variety of advantages in studying chemotaxis, including ease of growth, genetic tractability, and the conservation of mammalian signaling pathways. In this chapter, we describe the use of the under-agarose chemotaxis assay to understand the signaling pathways controlling directional sensing and motility in Dictyostelium discoideum. Given the similarities between Dictyostelium and mammalian cells, this allows us to dissect conserved pathways involved in eukaryotic chemotaxis.

Keywords: Amoeba; Chemotaxis; Dictyostelium; Folic acid; cAMP.

MeSH terms

  • Amoeba
  • Animals
  • Chemotaxis*
  • Cyclic AMP
  • Dictyostelium* / genetics
  • Pterins
  • Sepharose

Substances

  • Pterins
  • Sepharose
  • Cyclic AMP