The directed beating of motile cilia is a critical aspect of tissue function in a variety of developmental and physiological contexts including proper neural development, egg migration through the oviduct and mucus clearance in the respiratory tract. The loss of cilia motility results in a wide range of phenotypes including hydrocephaly, infertility, situs inversus, and respiratory dysfunction. We have developed the ciliated epithelium of Xenopus larval skin as a model system to ask: How do ciliated cells generate hundreds of cilia and how do they orient those cilia in an organized way? We have developed confocal light microscopic methods for visualizing specific aspects of ciliated cells in the developing skin of Xenopus embryos. These methods allow us to visualize the massive centriole duplication required to generate the approximately 100 basal bodies that nucleate the cilia. Additionally, we can visualize and accurately quantify the developmental process by which cilia orientation goes from weakly biased to precisely oriented, a process we call cilia refinement. Using these methods we will address: (1.) The dynamics of cilia refinement, (2.) The regulation of cilia refinement, and (3.) The regulation of centriole duplication. Our results will provide useful information regarding the development of ciliated epithelia. Additionally, loss of cilia and ciliated cells, followed by regrowth, occurs in mature ciliated epithelia in response to transient events including chemical insult (e.g. smoke inhalation), natural processes (e.g. menstrual cycle), and disease (e.g. asthma). Results from our experiments will provide a model for understanding the homeostasis of ciliated cell polarity that is relevant for designing novel therapies directed towards accelerating the rate of cilia reorientation after respiratory stress.

Public Health Relevance

The ability to generate directed fluid flow is critical to human health particularly in the respiratory system and the female reproductive tract. The goal of this project is to understand how any organ develops the cellular structures called cilia that are required to generate this flow. Specifically, we are interested in how cells generate hundreds of cilia, how these cilia become polarized and how they maintain this polarity under challenges such as respiratory disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM089970-05
Application #
8638975
Study Section
Development - 1 Study Section (DEV1)
Program Officer
Hoodbhoy, Tanya
Project Start
2010-04-02
Project End
2015-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
5
Fiscal Year
2014
Total Cost
$350,188
Indirect Cost
$121,030
Name
Northwestern University Chicago
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
Kim, Sun K; Zhang, Siwei; Werner, Michael E et al. (2018) CLAMP/Spef1 regulates planar cell polarity signaling and asymmetric microtubule accumulation in the Xenopus ciliated epithelia. J Cell Biol 217:1633-1641
Werner, Michael; Del Castillo, Urko; Ventrella, Rosa et al. (2018) The small molecule AMBMP disrupts microtubule growth, ciliogenesis, cell polarity, and cell migration. Cytoskeleton (Hoboken) 75:450-457
Galati, Domenico F; Mitchell, Brian J; Pearson, Chad G (2016) Subdistal Appendages Stabilize the Ups and Downs of Ciliary Life. Dev Cell 39:387-389
Silva, Erica; Betleja, Ewelina; John, Emily et al. (2016) Ccdc11 is a novel centriolar satellite protein essential for ciliogenesis and establishment of left-right asymmetry. Mol Biol Cell 27:48-63
Vladar, Eszter K; Mitchell, Brian J (2016) It's a family act: the geminin triplets take center stage in motile ciliogenesis. EMBO J 35:904-6
Jaffe, Kimberly M; Grimes, Daniel T; Schottenfeld-Roames, Jodi et al. (2016) c21orf59/kurly Controls Both Cilia Motility and Polarization. Cell Rep 14:1841-9
Wong, Yao Liang; Anzola, John V; Davis, Robert L et al. (2015) Cell biology. Reversible centriole depletion with an inhibitor of Polo-like kinase 4. Science 348:1155-60
Zhang, Siwei; Mitchell, Brian J (2015) Centriole biogenesis and function in multiciliated cells. Methods Cell Biol 129:103-127
Werner, Michael E; Mitchell, Jennifer W; Putzbach, William et al. (2014) Radial intercalation is regulated by the Par complex and the microtubule-stabilizing protein CLAMP/Spef1. J Cell Biol 206:367-76
Chien, Yuan-Hung; Werner, Michael E; Stubbs, Jennifer et al. (2013) Bbof1 is required to maintain cilia orientation. Development 140:3468-77

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