Efficient spatiotemporal analysis of the flagellar waveform of Chlamydomonas reinhardtii.
Efficient spatiotemporal analysis of the flagellar waveform of Chlamydomonas reinhardtii.
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DOI：
10.1002/cm.20424
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发表时间：
2010-01
期刊：
影响因子：
2.9
通讯作者：
Dutcher, S. K.
中科院分区：
文献类型：
--
作者：
Bayly, P. V.;Lewis, B. L.;Kemp, P. S.;Pless, R. B.;Dutcher, S. K.
9 + 2轴丝是一个微管为基础的机器，为纤毛和鞭毛的振荡跳动提供动力。其高度调节的运动对于许多器官的正常功能是必不可少的;纤毛病导致先天性缺陷、慢性呼吸道感染和不育。我们提出了一种有效的方法来获得鞭毛运动的定量描述，具有高的空间和时间分辨率，从高速视频记录的明场图像。这种高度自动化的技术提供了形状，剪切角，曲率，和弯曲传播速度沿着鞭毛的长度，与每拍约200个时间样本。我们比较了单鞭毛野生型和ida 3突变细胞的波形，缺乏I1内部动力蛋白复合物。视频图像以350 fps捕获。通过基于快速傅立叶变换（FFT）的配准消除刚体运动，并确定鞭毛上点的笛卡尔（x-y）坐标。这些x-y“点云”使用Isomap（一种非线性降维方法）嵌入两个数据维度中，并按鞭毛周期中的阶段进行排序。一个光滑的表面被拟合到排序的点云，这提供了高分辨率的剪切角和曲率的估计。野生型和ida 3细胞在剪切振幅上表现出很大的差异，但最大和最小曲率值相似。与野生型细胞相比，在ida 3细胞中，反向弯曲开始得更早，并且相对于主弯曲行进得更慢。鞭毛运动的调节必须涉及I1动力蛋白的方式与这些结果一致。
The 9 + 2 axoneme is a microtubule-based machine that powers the oscillatory beating of cilia and flagella. Its highly regulated movement is essential for the normal function of many organs; ciliopathies cause congenital defects, chronic respiratory tract infections and infertility. We present an efficient method to obtain a quantitative description of flagellar motion, with high spatial and temporal resolution, from high speed video recording of bright field images. This highly automated technique provides the shape, shear angle, curvature, and bend propagation speeds along the length of the flagellum, with ∼200 temporal samples per beat. We compared the waveforms of uniflagellated wild-type and ida3 mutant cells, which lack the I1 inner dynein complex. Video images were captured at 350 fps. Rigid-body motion was eliminated by fast Fourier transform (FFT)-based registration, and the Cartesian (x–y) coordinates of points on the flagellum were identified. These x–y “point clouds” were embedded in two data dimensions using Isomap, a nonlinear dimension reduction method, and sorted by phase in the flagellar cycle. A smooth surface was fitted to the sorted point clouds, which provides high-resolution estimates of shear angle and curvature. Wild-type and ida3 cells exhibit large differences in shear amplitude, but similar maximum and minimum curvature values. In ida3 cells, the reverse bend begins earlier and travels more slowly relative to the principal bend, than in wild-type cells. The regulation of flagellar movement must involve I1 dynein in a manner consistent with these results.
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DOI：
10.1083/jcb.92.3.722
发表时间：
1982-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Brokaw CJ;Luck DJ;Huang B
通讯作者：
Huang B
DOI：
10.1083/jcb.112.3.441
发表时间：
1991-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kamiya R;Kurimoto E;Muto E
通讯作者：
Muto E
影响因子：
--
作者：
BROKAW, CJ;LUCK, DJL
通讯作者：
LUCK, DJL
影响因子：
64.5
作者：
HUANG, B;RAMANIS, Z;LUCK, DJL
通讯作者：
LUCK, DJL
影响因子：
--
作者：
RUFFER, U;NULTSCH, W
通讯作者：
NULTSCH, W