DOI: 10.5281/zenodo.13881396
title: Active fluctuations of axoneme oscillations scale with number of dynein motors

description:  

The dataset published here was used to measure the phase fluctuations (frequency jitter) of isolated and reactivated axonemes from Chlamydomonas reinhardtii as a function of the dynein motor density in these axonemes. The corresponding article is published in PNAS: https://doi.org/10.1073/pnas.2406244121

Axonemes were isolated from wt(cc-125) and oda1(cc-2228) cells.

Reactivation was performed:(1) after dynein extraction with different amounts of salt (KCL) or(2) in the presence of different concentrations of ATP .

For all experimental conditions we provide the following data files:

Movies of reactivated axonemes:

Reactivated axonemes were imaged with phase-contrast microscopy. Recorded movies are organized in folders (.zip) which are labeled according to the respective reactivation condition (KCL and ATP concentration as well as the Chlamydomonas strain, from which the axonemes were purified, e.g. 50mM_KCl_750uM_ATP_ODA_Extracted.zip).

Those folders contain (1) movie files (multilayer .tif) of single axonemes and (.txt) files with a corresponding number-label (with microscope, camera settings and experimental condition (Chlamydomonas strain, [ATP], [KCL]).

Using the number-label, the corresponding data file can be identified.  

Data files (for the corresponding movies):

The experimental data is organized in MATLAB (.mat) files. Those files contain shape and waveform information (see below) for the respective reactivation condition (axoneme type, [ATP], [KCL] detailed below the file name).

WT_KCL_master.mat - WT axonemes, KCL extracted and reactivated with 750uM ATP

0mM_KCl_750uM_ATP_WT_Extracted50mM_KCl_750uM_ATP_WT_Extracted100mM_KCl_750uM_ATP_WT_Extracted200mM_KCl_750uM_ATP_WT_Extracted300mM_KCl_750uM_ATP_WT_Extracted400mM_KCl_750uM_ATP_WT_Extracted

WT_ATP_master.mat  - WT axonemes reactivated with different ATP concentraions

0mM_KCl_50uM_ATP_WT0mM_KCl_100uM_ATP_WT0mM_KCl_370uM_ATP_WT0mM_KCl_500uM_ATP_WT0mM_KCl_750uM_ATP_WT

ODA_KCL_master.mat - ODA axonemes, KCL extracted and reactivated with 750uM ATP

0mM_KCl_750uM_ATP_ODA_Extracted50mM_KCl_750uM_ATP_ODA_Extracted100mM_KCl_750uM_ATP_ODA_Extracted200mM_KCl_750uM_ATP_ODA_Extracted300mM_KCl_750uM_ATP_ODA_Extracted

ODA_ATP_master.mat - ODA axonemes reactivated with different ATP concentraions

0mM_KCl_70uM_ATP_ODA0mM_KCl_100uM_ATP_ODA0mM_KCl_370uM_ATP_ODA0mM_KCl_500uM_ATP_ODA0mM_KCl_750uM_ATP_ODA

Each file includes a data-cell ‘Master’ with a columns for the different experimental conditions (e.g. concentrations of ATP or KCL). Data-cells contain one structure for each axoneme. 

 

These structures have the following fields:

nframe…                                  number of frames of the dataset

phi_in_rad…                            phase angle (beat-cycle-phase) (rad)

tangent_angle_psi_in_rad…   the tangent angle for 24 positions along arc-length (rad)

xy_in_micron…                       the x,y positions (µm)

dt_in_second…                       time between adjacent frames (second)

ds_in_micron…                       spacing between adjacent arc-length positions (µm)

f0_in_Hz…                               beat frequency (Hz)

A…                                           beat amplitude (arc-length average) (rad)

Q…                                           quality factor

sexp…                                      string with experiment label

KCL_in_mM…                          KCL concentration (mM) used for dynein extraction

ATP_in_uM…                           ATP concentration (mM) used for reactivation

File…                                        filename, includes a number-label that corresponds to the imaging data provided