A Plasmodium plasma membrane reporter reveals membrane dynamics by live-cell microscopy

Abstract

During asexual replication within the Anopheles mosquito and their vertebrate host, Plasmodium parasites depend on the generation of a massive amount of new plasma membrane to produce thousands of daughter parasites. How the parasite plasma membrane (PPM) is formed has mostly been studied by electron microscopy, which does not allow an insight into the dynamics of this process. We generated a Plasmodium berghei reporter parasite line by GFP-tagging of a non-essential PPM-localized protein, and followed plasma membrane development in living parasites through the entire Plasmodium life cycle. By generating double-fluorescent parasites in which the PPM is visualized in combination with the parasite endoplasmic reticulum, we show that membrane contact sites are formed between both membrane systems during oocyst and liver stage development that might be used to deliver lipids to the dramatically expanding PPM. In conclusion, we have established a powerful tool to follow PPM development in living parasites, which promises to greatly expand our knowledge of membrane biology in the Plasmodium parasite.

Microtubule number and length determine cellular shape and function in Plasmodium

“…Strikingly, co‐labelling with the DNA stain Hoechst and our TEM images consistently showed that nuclear division and alignment at the periphery of the sporoblast (Burda et al , ) appeared initially not affected in α1‐tubulin (‐) parasites (Figs E and ) suggesting that chromosome segregation could proceed in the absence of microtubules, a phenomenon only reported once before for eukaryotic cells in fission yeast (Castagnetti et al , ). To investigate whether microtubules were absent in early stages of nuclear division within oocysts, we imaged oocysts from day 4 after infection.…”

BioID Reveals Novel Proteins of the Plasmodium Parasitophorous Vacuole Membrane

“…Instead, we decided to include liver stage parasites in our analysis, where such discrimination by IFA is possible. The cytomere stage is particularly suited to localization studies, since parasites are very large at that time point and the PPM has already started to invaginate ( 19 ), allowing a clear differentiation between a potential PVM and PV/PPM localization. We therefore concentrated on fully developed cytomere liver stage parasites, which were fixed and stained with an antiserum against EXP1 to perform colocalization analysis by confocal microscopy.…”

Transcriptome analysis ofPlasmodium bergheiduring exo-erythrocytic development

“…Each exo-erythrocytic stage parasite (EEF: exo-erythrocytic form) generates tens of thousands of nuclei by the process of exo-erythrocytic schizogony. This rapid nuclear division is accompanied by growth and replication of organelles including the Golgi apparatus, endoplasmic reticulum, mitochondrion and apicoplast and by the vast expansion of the plasma membrane (4–6). Nuclei and organelles are eventually segregated into individual merozoites.…”