Studying cilia in zebrafish.
Zebrafish are ideally suited for analysis of genes required for ciliogenesis and cilia function. Combining genetic manipulation with high-quality in vivo imaging, zebrafish embryos provide a high-throughput system for annotation of the cilia proteome. The specific advantages of the system are the availability of cilia mutants, the ability to target genes of unknown function using antisense methods, the feasibility of observing cilia in living embryos, and the ability to image fixed cilia in wholemount at high resolution. Techniques are described for analysis of mutants, gene knockdown using antisense morpholino oligos, visualizing cilia and cilia orientation in wholemount zebrafish embryos, cilia imaging by high-speed video, and electron microscopy of zebrafish cilia.
Drummond
Iain
I
Department of Medicine, Harvard Medical School and Massachusetts General Hospital,Charlestown, Massachusetts 02129, USA.
D000818:Animals; D030801:Animals, Genetically Modified; D002923:Cilia / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D018715:Microscopy, Video / Q000379:methods; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000033:anatomy & histology* / Q000196:embryology / Q000235:genetics / Q000378:metabolism
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger