Paramecium swimming and ciliary beating patterns: a study on four RNA interference mutations.
Paramecium cells swim and feed by beating their thousands of cilia in coordinated patterns. The organization of these patterns and its relationship with cell motility has been the subject of a large body of work, particularly as a model for ciliary beating in human organs where similar organization is seen. However the rapid motion of the cells makes quantitative measurements very challenging. Here we provide detailed measurements of the swimming of Paramecium cells from high-speed video at high magnification, as they move in microfluidic channels. An image analysis protocol allows us to decouple the cell movement from the motion of the cilia, thus allowing us to measure the ciliary beat frequency (CBF) and the spatio-temporal organization into metachronal waves along the cell periphery. Two distinct values of the CBF appear at different regions of the cell: most of the cilia beat in the range of 15 to 45 Hz, while the cilia in the peristomal region beat at almost double the frequency. The body and peristomal CBF display a nearly linear relation with the swimming velocity. Moreover the measurements do not display a measurable correlation between the swimming velocity and the metachronal wave velocity on the cell periphery. These measurements are repeated for four RNAi silenced mutants, where proteins specific to the cilia or to their connection to the cell base are depleted. We find that the mutants whose ciliary structure is affected display similar swimming to the control cells albeit with a reduced efficiency, while the mutations that affect the cilia's anchoring to the cell lead to strongly reduced ability to swim. This reduction in motility can be related to a loss of coordination between the ciliary beating in different parts of the cell.
Department of Mechanics, LadHyX, Ecole Polytechnique-CNRS, F-91128 Palaiseau, France. anette.funfak@espci.fr.
Anette
A
Funfak
Cathy
C
Fisch
Hatem T
HT
Abdel Motaal
Julien
J
Diener
Laurent
L
Combettes
Charles N
CN
Baroud
Pascale
P
Dupuis-Williams
D001683:Biological Clocks / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D002923:Cilia / Q000502:physiology* / Q000648:ultrastructure; D018715:Microscopy, Video / Q000379:methods; D020409:Molecular Motor Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009991:Oscillometry / Q000379:methods; D010247:Paramecium / Q000166:cytology* / Q000502:physiology*; D034622:RNA Interference / Q000502:physiology; D013550:Swimming / Q000502:physiology*
D020409:Molecular Motor Proteins