Chemical Screening for Flagella-Associated Phenotypes in Chlamydomonas reinhardtii
- 873 Downloads
Flagella of the unicellular green alga Chlamydomonas reinhardtii are nearly identical to cilia of vertebrate cells and provide an excellent model to study ciliogenesis. Cilia and flagella are important organelles used for motility and sensing the extracellular environment. Abnormalities in cilia structure or ciliary dysfunction can have devastating consequences ranging from diabetes and obesity to polycystic kidney disease and mental retardation. Small-molecule inhibitor libraries can be used to screen for flagellum-associated phenotypes in assembly, length, motility, deflagellation, and cellular toxicity. These phenotypes can be assessed from direct microscopic visualization and custom-designed assays. These methods identify fundamental regulators of ciliary biology as well as potential therapeutic interventions for ciliopathies.
Key wordsChemical screen Motility Chlamydomonas reinhardtii Flagellar length
We would like to thank all the members of the Avasthi Lab for the critical reading of the manuscript. This work was funded by P20 GM104936-09 (PA) and NSF GRFP 1518767 (BJ). Thanks to Soumita Dutta for providing images of healthy and contaminated cells for Fig. 1.
- 2.Cole DG, Diener DR, Himelblau AL, Beech PL, Fuster JC, Rosenbaum JL (1998) Chlamydomonas kinesin-II-dependent intraflagellar transport (IFT): IFT particles contain proteins required for ciliary assembly in Caenorhabditis elegans sensory neurons. J Cell Biol 141:993–1008CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Avasthi P, Marley A, Lin H, Gregori-Puigjane E, Shoichet BK, von Zastrow M et al (2012) A chemical screen identifies class a g-protein coupled receptors as regulators of cilia. Am Soc Chem Biol 7:911–919Google Scholar
- 7.Hutner SH, Provasoli L, Scahtz A, Haskins P (1950) Some approaches to the study of the role of metals in the metabolism of microorganisms. Proc Am Philos Soc 94:152–170Google Scholar
- 9.Kozminski KG, Beech PL, Rosenbaum JL (1995) The Chlamydomonas kinesin-like protein FLA10 is involved in motility associated with the flagellar membrane. J Cell Biol 12:222–234Google Scholar
- 16.Dutta S, Avasthi P (2017) Flagellar synchronization is a simple alternative to cell cycle synchronization for ciliary and flagellar studies. mSphere 2(2). https://doi.org/10.1128/mSphere.00003-17
- 17.Sato M, Murata Y, Mizusawa M, Iwashashi H, Shu-ichi O (2004) A simple and rapid dual-fluorescence viability assay for microalgae. Micrbiol Cult Coll 20:53–59Google Scholar