Detail of Fig4B_DuringBend
Project
SSBD:Repository
Title
Calcium imaging of immotile cilia and crown cells of Inversin mutant embryo in the presence of BMP4 during manipulation of cilia using optical tweezer
Description
Calcium imaging of immotile cilia and crown cells of iv/iv mouse embryo in the presence of BMP4 during manipulation of cilia using optical tweezer. GCaMP6 was expressed in the cytoplasm for cytoplasmic calcium imaging. Immotile cilia at the node were visualized based on mCherry expression. From T=96 to T=908 of imaging, the immotile cilia was bent to the ventral side using optical tweezer. From T=1012 to T=1825 of imaging, the immotile cilia was bent to the dorsal side using the tweezer.
Release, Updated
2025-11-28
License
CC BY
Kind
Image data
File Formats
.tif
Data size
3.9 GB
Organism
Mus musculus
(
NCBI:txid10090
)
Strain(s)
Dnah11 tm1Bknr, B6-Tg(Nodal NDE-5HT6/GCaMP6,5HT6/mCherry)35Hmd, STOCK Tg(Nodal NDE-GCaMP6)29AHmd
Cell Line
-
Datatype
-
Molecular Function (MF)
Biological Process (BP)
calcium-mediated signaling
Cellular Component (CC)
non-motile cilium
Biological Imaging Method
time lapse microscopy
(
Fbbi:00000249
)
X scale
0.43 micrometer
Y scale
0.43 micrometer
Z scale
-
T scale
0.303 seconds (5-frame running average)
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-
Summary of Methods
Katoh TA, Lange T, Nakajima Y, Yashiro K, Okada Y, Hamada H. BMP4 regulates asymmetric Pkd2 distribution in mouse nodal immotile cilia and ciliary mechanosensing required for left-right determination. Dev Dyn. 2024 Jul 10.
Related paper(s)
Takanobu A Katoh, Tim Lange, Yoshiro Nakajima, Kenta Yashiro, Yasushi Okada, Hiroshi Hamada (2024) BMP4 regulates asymmetric Pkd2 distribution in mouse nodal immotile cilia and ciliary mechanosensing required for left-right determination., Developmental dynamics : an official publication of the American Association of Anatomists
Published in 2024 Jul 10 (Electronic publication in July 10, 2024, midnight )
- doi: 10.1002/dvdy.727
-
pmid: 38984461
(Abstract) BACKGROUND: Mouse nodal immotile cilia mechanically sense the bending direction for left-right (L-R) determination and activate the left-side-specific signaling cascade, leading to increased Nodal activity. Asymmetric distribution of Pkd2, a crucial channel for L-R determination, on immotile cilia has been reported recently. However, the causal relationship between the asymmetric Pkd2 distribution and direction-dependent flow sensing is not well understood. Furthermore, the underlying molecular mechanism directing this asymmetric Pkd2 distribution remains unclear. RESULTS: The effects of several recombinant proteins and inhibitors on the Pkd2 distribution were analyzed using super-resolution microscopy. Notably, bone morphogenetic protein 4 (BMP4) affected the Pkd2 distribution. Additionally, three-dimensional manipulation of nodal immotile cilia using optical tweezers revealed that excess BMP4 caused defects in the mechanosensing ability of the cilia. CONCLUSIONS: Experimental data together with model calculations suggest that BMP4 regulates the asymmetric distribution of Pkd2 in nodal immotile cilia, thereby affecting the ability of these cilia to sense the bending direction for L-R determination. This study, for the first time, provides insight into the relationship between the asymmetric protein distribution in cilia and their function.
Contact
Contributors
OMERO Project