Imaging cilia in zebrafish.
Research focused on cilia as extremely important cellular organelles has flourished in recent years. A thorough understanding of cilia regulation and function is critical, as disruptions of cilia structure and/or function have been linked to numerous human diseases and disorders. The tropical freshwater zebrafish is an excellent model organism in which to study cilia structure and function. We can readily image cilia and their motility in embryonic structures including Kupffer's vesicle during somite stages and the pronephros from 1 day postfertilization onward. Here, we describe how to image cilia by whole-mount immunofluorescence, transverse cryosection/immunohistochemistry, and transmission electron microscopy. We also describe how to obtain videos of cilia motility in living embryos.
Jaffe
Kimberly M
KM
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
Thiberge
Stephan Y
SY
Bisher
Margaret E
ME
Burdine
Rebecca D
RD
D000818:Animals; D002923:Cilia / Q000737:chemistry / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure*; D016616:Cryoultramicrotomy / Q000379:methods; D004625:Embryo, Nonmammalian; D006801:Humans; D008853:Microscopy / Q000379:methods*; D008954:Models, Biological; D009068:Movement / Q000502:physiology; D015027:Zebrafish / Q000196:embryology / Q000502:physiology