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Hagiwara, Haruo; Kano, Akira; Aoki, Takeo; Ohwada, Nobuo; Takata, Kuniaki

doi:10.1023/a:1004163315822

Cited by 30 publications

(16 citation statements)

References 12 publications

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“…Galectin-3 was detected at the ciliary rootlet and at the basal foot cap (Figure 3C, black arrows and black arrowheads, respectively). Basal foot caps and ciliary rootlets have been suggested to be the MT anchoring sites of the BB, and are now considered as the MT organizing centers (MTOCs) of the cilium base 26, 27, 28. Consistently, immunogold staining revealed that γ-tubulin is located at the basal foot cap and at the ciliary rootlet in mouse tracheal multiciliated cells (Figure 3D, arrows and arrowheads, respectively), as previously observed in oviduct epithelial cells 27.…”

Section: Resultssupporting

confidence: 79%

“…Basal foot caps and ciliary rootlets have been suggested to be the MT anchoring sites of the BB, and are now considered as the MT organizing centers (MTOCs) of the cilium base 26, 27, 28. Consistently, immunogold staining revealed that γ-tubulin is located at the basal foot cap and at the ciliary rootlet in mouse tracheal multiciliated cells (Figure 3D, arrows and arrowheads, respectively), as previously observed in oviduct epithelial cells 27. Given Galectin-3 localization with the basal foot cap, we conclude that this lectin is associated with the cilium base MTOC.…”

Section: Resultsmentioning

confidence: 99%

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Basal foot MTOC organizes pillar MTs required for coordination of beating cilia

et al. 2014

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Coordination of ciliary beating is essential to ensure mucus clearance in the airway tract. The orientation and synchronization of ciliary motion responds in part to the organization of the underlying cytoskeletal networks. Using electron tomography on mouse trachea, we show that basal bodies are collectively hooked at the cortex by a regular microtubule array composed of 4-5 microtubules. Removal of Galectin-3, one of basal body components, provokes misrecruitment of γ-tubulin, disorganization of this microtubule framework emanating from the basal foot cap, together with loss of basal body alignment and cilium orientation, defects in cilium organization and reduced fluid flow in the tracheal lumen. We conclude that Galectin-3 plays a crucial role in the maintenance of the microtubule organizing center of the cilium and the “pillar” microtubules, and that this network is instrumental for the coordinated orientation and stabilization of motile cilia.

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Section: Resultssupporting

confidence: 79%

Section: Resultsmentioning

confidence: 99%

Basal foot MTOC organizes pillar MTs required for coordination of beating cilia

et al. 2014

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“…Either antibodies against known markers of the basal body and the basal foot or lentivirally expressed epitope-tagged transgenes can be used with the various super-resolution microscopy techniques. Here, we will focus on three-dimensional structured illumination microscopy (3D-SIM) immunofluorescence imaging of the transition fiber component Cep164 (Graser et al, 2007; Lau, Lee, Sahl, Stearns, & Moerner, 2012) and the basal foot component γ-tubulin (Hagiwara, Kano, Aoki, Ohwada, & Takata, 2000) (Figure 3). 3D-SIM requires minimal adjustment to established immunofluorescence protocols, while permitting super-resolution imaging in up to four channels.…”

Section: Assessment Of Ultrastructural Planar Polarity Of Motile Cmentioning

confidence: 99%

Observing planar cell polarity in multiciliated mouse airway epithelial cells

Vladar

Lee

Stearns

et al. 2015

Methods in Cell Biology

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The concerted movement of cilia propels inhaled contaminants out of the lungs, safeguarding the respiratory system from toxins, pathogens, pollutants, and allergens. Motile cilia on the multiciliated cells (MCCs) of the airway epithelium are physically oriented along the tissue axis for directional motility, which depends on the planar cell polarity (PCP) signaling pathway. The MCCs of the mouse respiratory epithelium have emerged as an important model for the study of motile ciliogenesis and the PCP signaling mechanism. Unlike other motile ciliated or planar polarized tissues, airway epithelial cells are relatively easily accessible and primary cultures faithfully model many of the essential features of the in vivo tissue. There is growing interest in understanding how cells acquire and polarize motile cilia due to the impact of mucociliary clearance on respiratory health. Here, we present methods for observing and quantifying the planar polarized orientation of motile cilia both in vivo and in primary culture airway epithelial cells. We describe how to acquire and evaluate electron and light microscopy images of ciliary ultrastructural features that reveal planar polarized orientation. Furthermore, we describe the immunofluorescence localization of PCP pathway components as a simple readout for airway epithelial planar polarization and ciliary orientation. These methods can be adapted to observe ciliary orientation in other multi- and monociliated cells and to detect PCP pathway activity in any tissue or cell type.

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“…Almost all cell types have a monocilium, a microtubule-based structure protruding into the extracellular space [49]. In the case of an epithelial cell, the primary cilium is localized in the apical surface.…”

Section: Function At the Primary Ciliummentioning

confidence: 99%

“…In the case of an epithelial cell, the primary cilium is localized in the apical surface. The cilium is formed from the basal body in growth arrested cells [49]. While the exact function of the immotile, primary cilium of kidney epithelial cells is not well understood, it is now well established that loss of ciliary function and/or formation results in cystic diseases pointing to a central role of this organelle in PKD pathophysiology [50,51].…”

Section: Function At the Primary Ciliummentioning

confidence: 99%

Cell biology of polycystin-2

Tsiokas¹,

Kim²,

Ong³

2007

Cellular Signalling

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Naturally occurring mutations in two separate, but interacting loci, pkd1 and pkd2 are responsible for almost all cases of autosomal dominant polycystic kidney disease (ADPKD). ADPKD is one of the most common genetic diseases resulting primarily in the formation of large kidney, liver, and pancreatic cysts. Homozygous deletion of either pkd1 or pkd2 results in embryonic lethality in mice due to kidney and heart defects illustrating their indispensable roles in mammalian development. However, the mechanism by which mutations in these genes cause ADPKD and other developmental defects are unknown. Research in the past several years has revealed that PKD2 has multiple functions depending on its subcellular localization. It forms a receptor-operated, non-selective cation channel in the plasma membrane, a novel intracellular Ca 2+ release channel in the endoplasmic reticulum (ER), and a mechanosensitive channel in the primary cilium. This review focuses on the functional compartmentalization of PKD2, its modes of activation, and PKD2-mediated signal transduction.

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Cited by 30 publications

References 12 publications

Basal foot MTOC organizes pillar MTs required for coordination of beating cilia

Basal foot MTOC organizes pillar MTs required for coordination of beating cilia

Observing planar cell polarity in multiciliated mouse airway epithelial cells

Cell biology of polycystin-2

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