Disruption of TgPHIL1 alters specific parameters of Toxoplasma gondii motility measured in a quantitative, three-dimensional live motility assay.
Disruption of TgPHIL1 alters specific parameters of Toxoplasma gondii motility measured in a quantitative, three-dimensional live motility assay.
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DOI：
10.1371/journal.pone.0085763
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Ward GE
中科院分区：
文献类型：
--
作者：
Leung JM;Rould MA;Konradt C;Hunter CA;Ward GE
T.弓形虫利用底物依赖性滑动运动侵入其宿主细胞，在其裂解周期结束时从这些细胞中排出，并在感染期间通过宿主生物体传播。因此，寄生虫的移动能力对其毒力至关重要。T.刚地虫在有涂层的二维表面上进行三种不同类型的滑动运动：旋转、圆形滑动和螺旋形滑动。我们在这里表明，运动在一个三维的基质胶为基础的环境是惊人的不同，在所有的寄生虫移动不规则的螺旋状轨迹。为定量分析运动参数而开发的方法沿着平滑轨迹显示了复杂但周期性的运动模式，平均和最大速度分别为0.58±0.07 µm/s和2.01±0.17 µm/s。为了测试寄生虫新月形的变化如何影响轨迹参数，我们将Δ phil 1寄生虫的运动性与相应的亲本和互补系进行了比较，Δ phil 1寄生虫比野生型更短更宽。尽管所有三个品系的寄生虫移动百分比相当，但与亲本寄生虫品系相比，Δ phil 1突变体表现出显著降低的轨迹长度以及平均和最大速度。这些效应在Δ phil 1突变体的互补作用下部分或完全恢复。这些结果表明，形态的改变可能对T。弓形虫在细胞外基质样环境中的运动性，为Δ phil 1寄生虫体内适应性降低提供了可能的解释，并证明了定量三维测定用于研究寄生虫运动性的实用性。
T. gondii uses substrate-dependent gliding motility to invade cells of its hosts, egress from these cells at the end of its lytic cycle and disseminate through the host organism during infection. The ability of the parasite to move is therefore critical for its virulence. T. gondii engages in three distinct types of gliding motility on coated two-dimensional surfaces: twirling, circular gliding and helical gliding. We show here that motility in a three-dimensional Matrigel-based environment is strikingly different, in that all parasites move in irregular corkscrew-like trajectories. Methods developed for quantitative analysis of motility parameters along the smoothed trajectories demonstrate a complex but periodic pattern of motility with mean and maximum velocities of 0.58±0.07 µm/s and 2.01±0.17 µm/s, respectively. To test how a change in the parasite's crescent shape might affect trajectory parameters, we compared the motility of Δphil1 parasites, which are shorter and wider than wild type, to the corresponding parental and complemented lines. Although comparable percentages of parasites were moving for all three lines, the Δphil1 mutant exhibited significantly decreased trajectory lengths and mean and maximum velocities compared to the parental parasite line. These effects were either partially or fully restored upon complementation of the Δphil1 mutant. These results show that alterations in morphology may have a significant impact on T. gondii motility in an extracellular matrix-like environment, provide a possible explanation for the decreased fitness of Δphil1 parasites in vivo, and demonstrate the utility of the quantitative three-dimensional assay for studying parasite motility.
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DOI：
10.1084/jem.20020258
发表时间：
2002-06-17
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Barragan A;Sibley LD
通讯作者：
Sibley LD
影响因子：
48
作者：
Andenmatten, Nicole;Egarter, Saskia;Jackson, Allison J;Jullien, Nicolas;Herman, Jean-Paul;Meissner, Markus
通讯作者：
Meissner, Markus
影响因子：
--
作者：
Donald, RGK;Allocco, J;Liberator, PA
通讯作者：
Liberator, PA
影响因子：
3.3
作者：
Håkansson, S;Morisaki, H;Sibley, LD
通讯作者：
Sibley, LD
影响因子：
4.7
作者：
Hegge, Stephan;Kudryashev, Mikhail;Frischknecht, Friedrich
通讯作者：
Frischknecht, Friedrich