Use of a highly specific kinase inhibitor for rapid, simple and precise synchronization of Plasmodium falciparum and Plasmodium knowlesi asexual blood-stage parasites.

PloS One
M. RessurreicaoChristiaan van Ooij

Abstract

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 egress from the infected 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 within a window of one to several 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 so that all parasites are within a window of development of several minutes, with a simple wash step. Furthermore, we show that parasites remain viable for several hours after becoming arrested by the compound a...Continue Reading

References

Aug 20, 1976·Science·W Trager, J B Jensen
May 1, 1968·Proceedings of the Society for Experimental Biology and Medicine·C Y Chang, E Simon
May 1, 1983·The Journal of protozoology·E M RivadeneiraC T Espinal
Jan 1, 1982·Transactions of the Royal Society of Tropical Medicine and Hygiene·A Knight, R E Sinden
Aug 2, 2003·Science·Karine G Le RochElizabeth A Winzeler
Apr 24, 2004·Antimicrobial Agents and Chemotherapy·Martin SmilksteinMichael Riscoe
Aug 1, 1989·Parasitology Today·D Kwiatkowski, B M Greenwood
Nov 5, 2004·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Lawrence Howard BannisterGraham Howard Mitchell
Dec 6, 2005·Molecular and Biochemical Parasitology·Carmen A DiazPaul A Liberator
Feb 24, 2006·Nucleic Acids Research·Manuel LlinásJoseph L DeRisi
Jun 13, 2006·Molecular and Biochemical Parasitology·Robert G K DonaldPaul A Liberator
Dec 17, 2009·Nature Protocols·Azar RadfarJosé M Bautista
Jun 23, 2010·Malaria Journal·Lisa C Ranford-CartwrightJonathan M Mwangi
Jul 28, 2010·Proceedings of the National Academy of Sciences of the United States of America·Michelle J BoyleJames G Beeson
Jan 7, 2011·Proceedings. Biological Sciences·Aidan J O'DonnellSarah E Reece
Jan 27, 2011·Nature Communications·Christof GrüringTobias Spielmann
Dec 21, 2012·Trends in Parasitology·Nicole MideoC Jessica E Metcalf
Dec 26, 2012·Proceedings of the National Academy of Sciences of the United States of America·Robert W MoonMichael J Blackman
Mar 29, 2013·Nature Communications·Caeul LimManoj T Duraisingh
Aug 14, 2013·Cellular Microbiology·Ellen KnuepferAnthony A Holder
Jun 16, 2016·Proceedings of the National Academy of Sciences of the United States of America·Robert W MoonAnthony A Holder
Mar 16, 2017·Proceedings of the National Academy of Sciences of the United States of America·Victoria L HaleHelen R Saibil
Jul 27, 2017·Scientific Reports·Lee MurrayDavid J Conway
Sep 30, 2017·The Journal of Antimicrobial Chemotherapy·Donelly A van SchalkwykColin J Sutherland

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Citations

Mar 10, 2021·Journal of Cell Science·Michele S Y Tan, Michael J Blackman
Aug 22, 2021·Scientific Reports·Hassan HakimiShinichiro Kawazu

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Methods Mentioned

BETA
transgenic
flow cytometry
flow cyctometry

Software Mentioned

- Elements
Attune NxT Flow Cytometer
Nikon NIS
LifeArc
GraphPad Prism
ImageJ

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