Calcium Signaling throughout the Toxoplasma gondii Lytic Cycle A STUDY USING GENETICALLY ENCODED CALCIUM INDICATORS
Calcium Signaling throughout the Toxoplasma gondii Lytic Cycle A STUDY USING GENETICALLY ENCODED CALCIUM INDICATORS
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DOI：
10.1074/jbc.m115.652511
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发表时间：
2015-11-06
影响因子：
4.8
通讯作者：
Moreno, Silvia N. J.
中科院分区：
文献类型：
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作者：
Borges-Pereira, Lucas;Budu, Alexandre;Moreno, Silvia N. J.
背景：Ca ~（2+）信号在T细胞溶解周期中起重要作用。gondii.Results:遗传编码的Ca 2+指示剂揭示了细胞溶质Ca 2+在真实的时间内的变化。结论：新方法突出了溶解周期的重要特征。刚地弓形虫是一种专性细胞内寄生虫，侵入宿主细胞，产生一个寄生虫空泡，在那里它通过寄生虫空泡膜与宿主细胞胞质液连通。寄生虫的裂解周期开始于其离开宿主细胞，随后是滑行运动、锥形挤出、附着和侵入另一宿主细胞。在这里，我们报告说，Ca 2（+）振荡发生在寄生虫在出口，滑行，和入侵，这是裂解周期的关键步骤的胞质溶胶。细胞外Ca 2（+）增强了这些过程中的每一个。我们使用表达遗传编码的钙指示剂的速殖子克隆系与表达瞬时表达的不同颜色的钙指示剂的宿主细胞相结合，并且我们测量了寄生虫和宿主在排出过程中同时发生的Ca 2（+）变化。我们证明了宿主和寄生虫胞质Ca 2（+）振荡之间的联系。我们的方法还使我们能够测量运动寄生虫的两个新特征，这是由Ca 2（+）流入增强。这是第一项研究显示，实时，Ca 2（+）信号之前的出口和他们的直接联系与运动，一个重要的毒力性状。
Background: Ca2+ signaling is important for the lytic cycle of T. gondii.Results: Genetically encoded Ca2+ indicators revealed cytosolic Ca2+ changes in real time.Conclusion: New approach highlights important features of the lytic cycle.Significance: Ca2+ influx leads to signaling that results in enhancement of important lytic cycle features.Toxoplasma gondii is an obligate intracellular parasite that invades host cells, creating a parasitophorous vacuole where it communicates with the host cell cytosol through the parasitophorous vacuole membrane. The lytic cycle of the parasite starts with its exit from the host cell followed by gliding motility, conoid extrusion, attachment, and invasion of another host cell. Here, we report that Ca2(+) oscillations occur in the cytosol of the parasite during egress, gliding, and invasion, which are critical steps of the lytic cycle. Extracellular Ca2(+) enhances each one of these processes. We used tachyzoite clonal lines expressing genetically encoded calcium indicators combined with host cells expressing transiently expressed calcium indicators of different colors, and we measured Ca2(+) changes in both parasites and host simultaneously during egress. We demonstrated a link between cytosolic Ca2(+) oscillations in the host and in the parasite. Our approach also allowed us to measure two new features of motile parasites, which were enhanced by Ca2(+) influx. This is the first study showing, inreal time, Ca2(+) signals preceding egress and their direct link with motility, an essential virulence trait.