Analysis of properties of cilia using Tetrahymena thermophila.
Cilia and eukaryotic flagella are important structures required for the motility of cells, the movement of medium across the surfaces of cells, and the connections between the receptor and synthetic portions of sensory cells. The axoneme forms the cytoskeleton of the cilium comprising several hundreds of proteins that assemble into the 9 + 2 arrangement of outer doublet and central pair microtubules, the inner and outer rows of dynein arms, and many other structures. Tetrahymena thermophila is an excellent model organism for the study of cilia and ciliogenesis. The cell is covered by about 1,000 cilia which are essential for survival. Additionally, the Tetrahymena genome is available and targeted genetic manipulations are straightforward. In this chapter, we describe five protocols that examine properties of cilia: (a) measuring mRNA levels to see the effect of deciliation on gene expression; (b) swimming velocity and linearity; (c) ciliary length and density; (d) phagocytosis that occurs through the ciliated oral apparatus; and (e) depolarization-induced ciliary reversal.
Rajagopalan
Vidyalakshmi
V
Department of Biology, Indiana University-South Bend, South Bend, IN, USA.
Corpuz
Elizabeth O
EO
Hubenschmidt
Mark J
MJ
Townsend
Caroline R
CR
Asai
David J
DJ
Wilkes
David E
DE
D000818:Animals; D002923:Cilia / Q000235:genetics / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism; D015870:Gene Expression; D017125:Genes, Protozoan; D018715:Microscopy, Video; D010587:Phagocytosis / Q000235:genetics; D005082:Physical Exertion / Q000502:physiology; D012333:RNA, Messenger / Q000032:analysis; D016808:Tetrahymena thermophila / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism
D012333:RNA, Messenger; D014404:Tubulin; D004398:Dyneins