Use of a highly specific kinase inhibitor for rapid, simple and precise synchronization of Plasmodium falciparum and Plasmodium knowlesi asexual blood-stage parasites
Use of a highly specific kinase inhibitor for rapid, simple and precise synchronization of Plasmodium falciparum and Plasmodium knowlesi asexual blood-stage parasites
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使用高度特异性激酶抑制剂快速、简单且精确地同步恶性疟原虫和诺氏疟原虫无性血期寄生虫
DOI：
10.1101/2020.04.24.059493
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
C. van Ooij
中科院分区：
文献类型：
--
作者：
M. Ressurreição;James A. Thomas;S. Nofal;Christian Flueck;R. Moon;D. Baker;C. van Ooij
在红细胞性发育的无性阶段，疟原虫spp.寄生虫经历了一系列的形态变化，并诱导宿主细胞发生变化。在这一阶段结束时，寄生虫离开宿主细胞，之后后代入侵新的宿主细胞。这些过程是快速的，并且以依赖于时间的方式发生。尤其重要的是，在不同步的培养中，甚至在几个小时内同步的培养中，很难捕捉到寄生虫对红细胞的外泄和入侵。因此，寄生虫培养的精确同步对于研究这些过程是至关重要的。在这里，我们描述了一种使恶性疟原虫和诺氏疟原虫无性血液期寄生虫与ML10同步的方法，ML10是一种高度特异的cGMP依赖蛋白激酶(PKG)抑制剂，可在出口前大约15分钟阻止寄生虫的生长。这种抑制剂允许寄生虫培养同步到几分钟内，只需一个简单的清洗步骤。此外，我们表明，寄生虫在被该化合物阻止后仍能存活数小时，并且ML10比以前使用的PKG抑制剂化合物2具有优势。在这里，我们证明ML10是研究疟原虫种属的一个宝贵的工具。无性血液期生物学以及恶性疟原虫和诺氏疟原虫培养的常规同步。
During the course of the asexual erythrocytic stage of development, Plasmodium spp. parasites undergo a series of morphological changes and induce alterations in the host cell. At the end of this stage, the parasites exit the host cell, after which the progeny invade a new host cell. These processes are rapid and occur in a time-dependent manner. Of particular importance, egress and invasion of erythrocytes by the parasite are difficult to capture in an unsynchronized culture, or even a culture that has been synchronized to within hours. Therefore, precise synchronization of parasite cultures is of paramount importance for the investigation of these processes. Here we describe a method for synchronizing Plasmodium falciparum and Plasmodium knowlesi asexual blood stage parasites with ML10, a highly specific inhibitor of the cGMP-dependent protein kinase (PKG) that arrests parasite growth approximately 15 minutes prior to egress. This inhibitor allows parasite cultures to be synchronized to within minutes, with a simple wash step. Furthermore, we show that parasites remain viable for several hours after becoming arrested by the compound and that ML10 has advantages over the previously used PKG inhibitor Compound 2. Here, we demonstrate that ML10 is an invaluable tool for the study of Plasmodium spp. asexual blood stage biology and for the routine synchronization of P. falciparum and P. knowlesi cultures.
TNFα表达和食物摄入的每日节奏调节质量阶段与宿主昼夜节律的同步。
影响因子：
30.3
作者：
Hirako IC;Assis PA;Hojo-Souza NS;Reed G;Nakaya H;Golenbock DT;Coimbra RS;Gazzinelli RT
通讯作者：
Gazzinelli RT
恶性疟原虫：与 DL-α-二氟甲基鸟氨酸（一种多胺生物合成抑制剂）同步培养。
影响因子：
2.1
作者：
Assaraf,YG;Golenser,J;Spira,DT;Bachrach,U
通讯作者：
Bachrach,U