These authors have contributed equally to this work.
Electron tomography of Plasmodium falciparum merozoites reveals core cellular events that underpin erythrocyte invasion
Article first published online: 22 MAR 2013
© 2013 John Wiley & Sons Ltd
Volume 15, Issue 9, pages 1457–1472, September 2013
How to Cite
Hanssen, E., Dekiwadia, C., Riglar, D. T., Rug, M., Lemgruber, L., Cowman, A. F., Cyrklaff, M., Kudryashev, M., Frischknecht, F., Baum, J. and Ralph, S. A. (2013), Electron tomography of Plasmodium falciparum merozoites reveals core cellular events that underpin erythrocyte invasion. Cellular Microbiology, 15: 1457–1472. doi: 10.1111/cmi.12132
- Issue published online: 8 AUG 2013
- Article first published online: 22 MAR 2013
- Accepted manuscript online: 5 MAR 2013 08:16AM EST
- Manuscript Accepted: 27 FEB 2013
- Manuscript Revised: 25 FEB 2013
- Manuscript Received: 29 NOV 2012
- National Center for Research Resources. Grant Number: 5P41RR019664-08
- National Institute of General Medical Sciences. Grant Number: 8 P41 GM103445-08
- Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS
- National Health and Medical Research Council of Australia. Grant Numbers: NHMRC Project Grants 637340, APP1047085 JB & SAR
- The Australian Research Council. Grant Number: Discovery Project Grant DP120103161
- Human Frontier Science Organisation. Grant Number: HFSP YI Program Grant RGY0071/2011
- Australian Synchrotron and the Australian Academy of Science, the US Department of Energy, Office of Biological and Environmental Research. Grant Number: DE-AC02-05CH11231
- National Center for Research Resources of the National Institutes of Health. Grant Number: RR019664
- US Department of Energy, Office of Science
- University of Melbourne
- Australian Academy of Science
- OzeMalaR Network
- University of Heidelberg cluster of excellence CellNetworks
- Henning Stahlberg (University of Basel)
- Australian Research Council. Grant Numbers: FT100100112, FT0990350
Fig. S1. Sections from ROTO prepared tomograms of free intact Plasmodium falciparum merozoites.
A–C. Virtual sections through a merozoite showing connection between the inner membrane complex (IMC) and Endoplasmic reticulum (ER) membranes. Each image represents a 10 nm slice and each image is separated by 15 nm.
D–F. Virtual sections through a merozoite showing connection between the inner membrane complex (IMC) and Endoplasmic reticulum (ER) membranes. Scale bars are 200 nm.
Fig. S2. Cryo-electron tomography of a free intact Plasmodium falciparum merozoite.
A. Tomogram of a free intact Plasmodium falciparum merozoite. Note the discontinuities in the inner membrane complex (arrowheads).
B. Model constructed from CET of the apex of a free merozoite showing the distinct interruptions behind the apical third of the inner membrane complex (green). Rhoptries are shown in pink, apical rings are shown in yellow, microtubules are shown in dark blue, plasma membrane is shown light blue. Scale bars are 200 nm.
Fig. S3. Immunoelectron microscopy (IEM) of merozoites pre- and post-erythrocyte invasion. After sectioning, parasites are labelled with primary antibody then primary antibody is detected with a secondary antibody conjugated to colloidal gold (white arrows). Scale bars are 200 nm.
A. IEM of free merozoite (pre-invasion) labelled with the microneme (Mi) marker anti-AMA1.
B. IEM of free merozoite (pre-invasion) labelled with the rhoptry (Rh) maker anti-RAP1. IEM of merozoite (post-invasion) labelled with dense granule (Dg) marker anti-RESA.
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Movie S1. Electron tomography of a whole cryo-preserved Plasmodium falciparum merozoite. Cryo-electron tomogram of a free, intact Plasmodium falciparum merozoite. The tomogram was acquired on a FEI Titan Krios, and reconstructed with IMOD. Two rhoptries, numerous micronemes and two microtubules are visible at the apex of the merozoite. Note the opening in the inner membrane complex starting at the third of the apical rings.
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Movie S2. Serial electron tomogram of a ROTO-fixed free merozoite. Tomogram of a free Plasmodium falciparum merozoite reconstructed from serial sections. The tomogram was acquired on a Tecnai F30 (FEI, NL), reconstructed and segmented with IMOD. In the model, the nucleus is represented in red, the apicoplast in blue, the mitochondria in orange, the rhoptries in cyan and the dense granule and microneme are multicoloured. The continuum of the IMC/nuclear membrane and the ER is represented in gold.
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Movie S3. Serial electron tomogram of a GF-fixed merozoite with fusing rhoptries. Tomogram of the apex of an invading P. falciparum merozoite reconstructed from serial sections. The tomogram was acquired on a Tecnai F30 (FEI, NL), reconstructed with IMOD. The tomogram span 3 physical sections and details the entire volume of fusing rhoptries from the neck to the top of the bulbs.
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