Regulation of cilia abundance in multiciliated cells
- Washington University, United States
- Center for Cancer Research, National Cancer Institute, United States
- Stanford University, United States
Figures
Figure 1 with 1 supplement
Parental centrioles are dispensable for centriole amplification.
(a) Schematic of the proliferation and differentiation steps of multiciliated respiratory epithelia. The key stages of centriole amplification from parental centrioles and deuterosomes are depicted. …
Figure 1—video 1
Centrinone-mediated inhibition of Plk4 results in loss of parental centrioles at ALI0.
Samples were serially sectioned from at the cell’s apical surface until the basal membrane above the filter. The thickness of each section was 120 nm. Serial sections were imaged using a …
Figure 2 with 3 supplements
Loss of parental centrioles does not affect dynamics of centriole amplification.
(a) Immunofluorescence images of control and Centrinone-treated cells. Samples were fixed on the indicated days and stained for Plk4, centrioles (γ-tubulin) and apical cell-cell junctions (ZO-1). …
Figure 2—figure supplement 1
– Analysis of procentriole assembly in control and Centrinone-treated MTEC.
Immunofluorescence images of control and Centrinone-treated cells. Samples were fixed on the indicated days and stained for markers of parental centrioles and early procentriole nucleation as …
Figure 2—figure supplement 2
– Analysis of procentriole growth in control and Centrinone-treated MTEC.
Immunofluorescence images of control and Centrinone-treated cells. Samples were fixed on the indicated days and stained for markers of parental centrioles and procentriole growth as follows: (a) …
Figure 2—figure supplement 3
– Analysis of centriole maturation and ciliogenesis in control and Centrinone-treated MTEC.
(a) 3D-SIM images of MTEC at ALI0 and 12 stained for a marker of mature centrioles (distal appendage marker Cep164), centrioles (centrin), and apical cell junctions (ZO-1). Control cells at ALI0 …
Figure 3 with 1 supplement
Deuterosome biogenesis is unaffected by loss of parental centrioles.
(a) Immunofluorescence images of control and Centrinone-treated cells stained with antibodies against deuterosomes (Deup1), centrioles (γ-tubulin) and apical cell-cell junctions (ZO-1). Upper panels …
Figure 3—figure supplement 1
– Deuterosome-mediated procentriole formation is unaffected by loss of parental centrioles.
Quantification of procentriole number per deuterosome in control and Centrinone-treated cells at ALI3. We noted a slight but significant decrease in the average (a) and distribution (b) of …
Figure 4
Centriole abundance correlates with surface area and is unaffected by loss of PC.
(a) Example 3D-SIM images of mature multiciliated cells at ALI12 stained with markers of centrioles (centrin) and apical cell junctions (ZO-1). Scale bar = 10 μm. (b–c) Quantification of centriole …
Figure 5 with 1 supplement
Depletion of Plk4 causes a delay in centriologenesis but does not affect abundance.
(a) Images of MTEC infected with control or Plk4-targeting shRNA. Cells were fixed on the indicated days and stained for centrioles (centrin), endogenous Plk4, procentrioles (Sas6 or centrin), …
Figure 5—figure supplement 1
– Combined depletion and Centrinone-mediated inhibition of Plk4 causes a delay in centriologenesis.
(a) 3D-SIM images of MTEC infected with Plk4-targeting shRNA and treated with 1 μM Centrinone. Cells were fixed on the indicated days and stained for centrioles (centrin), endogenous Plk4 and apical …
Figure 6
Centriole abundance correlates with surface area in vivo.
(a) Example 3D-SIM image of adult mouse trachea (P60) immunostained for centrioles (centrin) and apical cell junctions (ZO-1). Scale bar = 10 µm. (b–d) Centriole number ranges between 100 and 400 …
Figure 7
Enlargement of the surface area of progenitor cells results in increased centriole abundance.
(a) Example 3D-SIM images of MTEC grown on varying concentrations of extracellular collagen matrix. Top panels show X-Y orientation, and lower panels the X-Z dimension. Scale bar = 10 µm. (b–e) …
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.44039.015
