Toxoplasma gondii pp 209-220 | Cite as
Phenotyping Toxoplasma Invasive Skills by Fast Live Cell Imaging
Abstract
Host cell invasion by Toxoplasma gondii/T. gondii tachyzoites is an obligate but complex multistep process occurring in second-scale. To capture the dynamic nature of the whole entry process requires fast and high-resolution live cell imaging. Recent advances in T. gondii/host cell genome editing and in quantitative live cell imaging—image acquisition and processing included—provide a systematic way to accurately phenotype T. gondii tachyzoite invasive behaviour and to highlight any variation or default from a standard scenario. Therefore, applying these combined strategies allows gaining deeper insights into the complex mechanisms underlying host cell invasion.
Key words
T. gondii tachyzoite Host cell invasion Live cell microscopy Image processingReferences
- 1.Tardieux I, Baum J (2016) Reassessing the mechanics of parasite motility and host-cell invasion. J Cell Biol 214:507–515. https://doi.org/10.1083/jcb.201605100CrossRefPubMedPubMedCentralGoogle Scholar
- 2.Sidik SM, Huet D, Ganesan SM, Huynh M-H, Wang T, Nasamu AS, Thiru P, Saeij JPJ, Carruthers VB, Niles JC, Lourido S (2016) A genome-wide CRISPR screen in Toxoplasma identifies essential apicomplexan genes. Cell 166:1423–1435.e12. https://doi.org/10.1016/j.cell.2016.08.019CrossRefPubMedPubMedCentralGoogle Scholar
- 3.Sidik SM, Huet D, Lourido S (2018) CRISPR-Cas9-based genome-wide screening of Toxoplasma gondii. Nat Protoc 13:307–323. https://doi.org/10.1038/nprot.2017.131CrossRefPubMedPubMedCentralGoogle Scholar
- 4.Bichet M, Joly C, Henni AH, Guilbert T, Xémard M, Tafani V, Lagal V, Charras G, Tardieux I (2014) The Toxoplasma-host cell junction is anchored to the cell cortex to sustain parasite invasive force. BMC Biol 12:773. https://doi.org/10.1186/s12915-014-0108-yCrossRefPubMedPubMedCentralGoogle Scholar
- 5.Hu K, Johnson J, Florens L, Fraunholz M, Suravajjala S, DiLullo C, Yates J, Roos DS, Murray JM (2006) Cytoskeletal components of an invasion machine--the apical complex of Toxoplasma gondii. PLoS Pathog 2:e13. https://doi.org/10.1371/journal.ppat.0020013CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Mondragon R, Frixione E (1996) Ca(2+)-dependence of conoid extrusion in Toxoplasma gondii tachyzoites. J Eukaryot Microbiol 43:120–127CrossRefGoogle Scholar
- 7.Lourido S, Tang K, Sibley LD (2012) Distinct signalling pathways control Toxoplasma egress and host-cell invasion. EMBO J 31:4524–4534. https://doi.org/10.1038/emboj.2012.299CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Lourido S, Moreno SNJ (2015) The calcium signaling toolkit of the apicomplexan parasites Toxoplasma gondii and Plasmodium spp. Cell Calcium 57:186–193. https://doi.org/10.1016/j.ceca.2014.12.010CrossRefPubMedGoogle Scholar
- 9.Carey KL, Westwood NJ, Mitchison TJ, Ward GE (2004) A small-molecule approach to studying invasive mechanisms of Toxoplasma gondii. Proc Natl Acad Sci U S A 101:7433–7438. https://doi.org/10.1073/pnas.0307769101CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Hakimi M-A, Olias P, Sibley LD (2017) Toxoplasma effectors targeting host signaling and transcription. Clin Microbiol Rev 30:615–645. https://doi.org/10.1128/CMR.00005-17CrossRefPubMedPubMedCentralGoogle Scholar
- 11.Graindorge A, Frénal K, Jacot D, Salamun J, Marq JB, Soldati-Favre D (2016) The conoid associated motor MyoH is indispensable for Toxoplasma gondii entry and exit from host cells. PLoS Pathog 12:e1005388. https://doi.org/10.1371/journal.ppat.1005388CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Mordue DG, Desai N, Dustin M, Sibley LD (1999) Invasion by Toxoplasma gondii establishes a moving junction that selectively excludes host cell plasma membrane proteins on the basis of their membrane anchoring. J Exp Med 190:1783–1792CrossRefGoogle Scholar
- 13.Charron AJ, Sibley LD (2004) Molecular partitioning during host cell penetration by Toxoplasma gondii. Traffic 5:855–867. https://doi.org/10.1111/j.1600-0854.2004.00228.xCrossRefPubMedGoogle Scholar
- 14.Suss-Toby E, Zimmerberg J, Ward GE (1996) Toxoplasma invasion: the parasitophorous vacuole is formed from host cell plasma membrane and pinches off via a fission pore. Proc Natl Acad Sci U S A 93:8413–8418CrossRefGoogle Scholar
- 15.Pavlou G, Biesaga M, Touquet B, Lagal V, Balland M, Dufour A, Hakimi M-A, Tardieux I (2018) Toxoplasma parasite twisting motion mechanically induces host cell membrane fission to complete invasion within a protective vacuole. Cell Host Microbe 24:81–96.e5. https://doi.org/10.1016/j.chom.2018.06.003CrossRefPubMedGoogle Scholar
- 16.Fox BA, Ristuccia JG, Gigley JP, Bzik DJ (2009) Efficient gene replacements in Toxoplasma gondii strains deficient for nonhomologous end joining. Eukaryot Cell 8:520–529. https://doi.org/10.1128/EC.00357-08CrossRefPubMedPubMedCentralGoogle Scholar
- 17.Huynh M-H, Carruthers VB (2009) Tagging of endogenous genes in a Toxoplasma gondii strain lacking Ku80. Eukaryot Cell 8:530–539. https://doi.org/10.1128/EC.00358-08CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Sidik SM, Hackett CG, Tran F, Westwood NJ, Lourido S (2014) Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9. PLoS One 9:e100450. https://doi.org/10.1371/journal.pone.0100450CrossRefPubMedPubMedCentralGoogle Scholar
- 19.Shen B, Brown KM, Lee TD, Sibley LD (2014) Efficient gene disruption in diverse strains of Toxoplasma gondii using CRISPR/CAS9. MBio 5:e01114–e01114. https://doi.org/10.1128/mBio.01114-14CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Andenmatten N, Egarter S, Jackson AJ, Jullien N, Herman J-P, Meissner M (2013) Conditional genome engineering in Toxoplasma gondii uncovers alternative invasion mechanisms. Nat Methods 10:125–127. https://doi.org/10.1038/nmeth.2301CrossRefPubMedGoogle Scholar
- 21.Bichet M, Touquet B, Gonzalez V, Florent I, Meissner M, Tardieux I (2016) Genetic impairment of parasite myosin motors uncovers the contribution of host cell membrane dynamics to Toxoplasma invasion forces. BMC Biol 14:97. https://doi.org/10.1186/s12915-016-0316-8CrossRefPubMedPubMedCentralGoogle Scholar
- 22.Rhee JM, Pirity MK, Lackan CS, Long JZ, Kondoh G, Takeda J, Hadjantonakis A-K (2006) In vivo imaging and differential localization of lipid-modified GFP-variant fusions in embryonic stem cells and mice. Genesis 44:202–218. https://doi.org/10.1002/dvg.20203CrossRefPubMedPubMedCentralGoogle Scholar
- 23.Egarter S, Andenmatten N, Jackson AJ, Whitelaw JA, Pall G, Black JA, Ferguson DJP, Tardieux I, Mogilner A, Meissner M (2014) The Toxoplasma Acto-MyoA motor complex is important but not essential for gliding motility and host cell invasion. PLoS One 9:e91819. https://doi.org/10.1371/journal.pone.0091819CrossRefPubMedPubMedCentralGoogle Scholar