Long-range migration of centrioles to the apical surface of the olfactory epithelium
- Department of Biology, Stanford University, United States
- Department of Genetics, Stanford University School of Medicine, United States
Figures
Figure 1 with 1 supplement
Centrioles in olfactory sensory neurons (OSNs) migrate tens of micrometers to the apical surface.
(A) Overview of the olfactory epithelium. Single-plane fluorescence image of a side view of the olfactory epithelium (left), corresponding to a schematic of cell types in the olfactory epithelium …
Figure 1—figure supplement 1
Centriole and cilium numbers in olfactory sensory neuron (OSN) progenitors and mature OSNs.
(A) Example of Imaris segmentation of centrioles in progenitor cells. (i) 3D image of centrioles in a progenitor cell. Green: staining for acetylated tubulin. (ii) The same image after segmentation …
Figure 2 with 2 supplements
Centrioles migrate in multiple groups during dendrite elongation.
(A) Within a single dendrite, centrioles migrate in multiple groups. Expansion microscopy – side view of expanded olfactory epithelium. In all side-view images, apical is oriented toward the top of …
-
Figure 2—source data 1
Migration rates of centriole groups from time-lapse imaging.
- https://cdn.elifesciences.org/articles/74399/elife-74399-fig2-data1-v2.xlsx
Figure 2—video 1
A centriole group migrates toward the apical surface.
Movie of the same centriole group as depicted in Figure 2C. Maximum x-projection image, showing a side view. White: eGFP-centrin2; blue: Hoechst, marking DNA in the most apical layer of nuclei, …
Figure 2—video 2
A centriole group with no net movement.
Movie of the same centriole group as depicted in Figure 2D. Maximum x-projection image, showing a side view. White: eGFP-centrin2; blue: Hoechst. Time between frames = 2 min.
Figure 3 with 1 supplement
Composition of centriole groups during olfactory sensory neuron (OSN) differentiation.
(A) The cohesion fiber and striated rootlet protein rootletin is absent during centriole migration but is gained at the mature dendritic knob. Expansion microscopy, maximum z-projection of confocal …
Figure 3—figure supplement 1
Expansion microscopy staining of rootletin in cycling cells.
Expansion microscopy – single-plane fluorescence image of a centrosome in expanded tissue culture cells (retinal pigment epithelium cells, RPE-1). Staining for rootletin (CROCC, cyan) demonstrates …
Figure 4 with 1 supplement
A single cilium forms prior to formation of multiple cilia.
(A) An olfactory sensory neuron (OSN) bearing a single cilium in the subapical compartment of the olfactory epithelium. Expansion microscopy – single-plane fluorescence image of a side view of the …
Figure 4—figure supplement 1
Centriole maturation in progenitors and migrating centriole groups.
(A) During centriole migration in immature olfactory sensory neurons (OSNs), a single centriole bears distal and subdistal appendages. Expansion microscopy – single-plane fluorescence image of …
Figure 5 with 1 supplement
Centriole progression toward the apical surface can be altered by stabilizing microtubules.
(A) Schematic: workflow from sample collection through imaging of olfactory epithelium explants. Septal olfactory epithelium was taken from adult mice expressing eGFP-centrin2 and Arl13b-mCherry and …
-
Figure 5—source data 1
Counts of centriole groups - paclitaxel treatment.
- https://cdn.elifesciences.org/articles/74399/elife-74399-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Migrating centriole groups in olfactory epithelium explants.
(A) Schematic diagram showing the locations of the subapical and middle/basal regions. Cilia are located in the apical region (above green region). The subapical compartment is located between the …
-
Figure 5—figure supplement 1—source data 1
Counts of centriole groups - explants prior to treatment.
- https://cdn.elifesciences.org/articles/74399/elife-74399-fig5-figsupp1-data1-v2.xlsx
-
Figure 5—figure supplement 1—source data 2
Counts of centriole groups - nocodazole treatment.
- https://cdn.elifesciences.org/articles/74399/elife-74399-fig5-figsupp1-data2-v2.xlsx
Figure 6
Summary of centriole migration and maturation during olfactory sensory neuron (OSN) differentiation.
Diagram summarizing the migration and maturation of centrioles over the course of OSN differentiation. A double solid line marks the basal lamina, and a dashed line is drawn above the apical …
Author response image 1
EB1 staining in cultured NIH3T3 cells and olfactory epithelium.
(A) Left: Mitotic 3T3 cell. Right: Interphase 3T3 cell. EB1 (green) and DAPI (blue). Both images are maximum z-projections of confocal stacks. (B) Olfactory epithelium. EB1 (white), β- tubulin III …
Tables
Table 1
Antibodies used in this study.
| Target | Source | Dilution |
|---|---|---|
| Acetylated tubulin | Sigma-Aldrich, clone 6-11B-1, RRID:AB_477585 | 1:1000 |
| CDK5RAP2 | Millipore, 06-1398 rabbit polyclonal, RRID:AB_11203651 | 1:200 |
| Centrin | EMD Millipore, clone 20H5, RRID:AB_10563501 | 1:1000 |
| CEP164 | Rabbit polyclonal previously described in Lee et al., 2014 | 1:500 |
| GFP | Rabbit antibody previously described in Hatch et al., 2010 | 1:2000 |
| GAP43 | Novus Biologicals, NB300, rabbit polyclonal, RRID:AB_921392 | 1:250 (Blocking buffer: PBSBT. 2% BSA, 0.1% Triton X-100, 1x PBS) |
| Pericentrin | BD Biosciences, clone 30, RRID:AB_399294 | 1:500 |
| Centriolin | Santa Cruz Biotech, clone C-9, RRID: AB_10851483 | 1:50 |
| STIL | Abcam, rabbit polyclonal, RRID:AB_2197878 | 1:1,000 |
| SASS6 | Santa Cruz Biotech, RRID:AB_1128357 | 1:200 |
| ODF2 | Novus Biologicals, mouse IgG2a, RRID:AB_1146453 | 1:200 |
| Rootletin (CROCC) | Santa Cruz Biotech, clone C-2, RRID:AB_10918081 | 1:100–200 |
| β-Tubulin III | BioLegend, clone TuJ1, RRID:AB_2313773 | 1:1000–2000 |
| Gamma-tubulin | Sigma-Aldrich, clone GTU88, RRID:AB_532292 | 1:1000 |
