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Article

Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets

Department of Biotechnology, University of Rijeka, 51000 Rijeka, Croatia
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Author to whom correspondence should be addressed.
Academic Editor: Friedrich Jung
Life 2021, 11(12), 1331; https://doi.org/10.3390/life11121331
Received: 10 November 2021 / Revised: 23 November 2021 / Accepted: 28 November 2021 / Published: 1 December 2021
(This article belongs to the Section Cell Biology and Tissue Engineering)
Phosphoinositides (PIs) are phosphorylated membrane lipids that have a plethora of roles in the cell, including vesicle trafficking, signaling, and actin reorganization. The most abundant PIs in the cell are phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] and phosphatidylinositol-4-monophosphate (PI4P). The localization and roles of both PI(4,5)P2 and PI4P are well established, is the broadly accepted methodological approach for their immunocytochemical visualization in different cell compartments in several cell lines. However, not much is known about these PIs in platelets (PLTs), the smallest blood cells that detect vessel wall injury, activate, and stop the bleeding. Therefore, we sought to investigate the localization of PI(4,5)P2 and PI4P in resting and activated PLTs by antibody staining. Here, we show that the intracellular pools of PI(4,5)P2 and PI4P can be detected by the established staining protocol, and these pools can be modulated by inhibitors of OCRL phosphatase and PI4KIIIα kinase. However, although resting PLTs readily stain for the plasma membrane (PM) pools of PI(4,5)P2 and PI4P, just a few activated cells were stained with the established protocol. We show that optimized protocol allows for the visualization of PI(4,5)P2 and PI4P at PM in activated PLTs, which could also be modulated by OCRL and PI4KIIIα inhibitors. We conclude that PI(4,5)P2 and PI4P are more sensitive to lipid extraction by permeabilizing agents in activated than in resting human PLTs, which suggests their different roles during PLT activation. View Full-Text
Keywords: platelets; phosphatidylinositol-4,5-bisphosphate; phosphatidylinositol-4-monophosphate; immunofluorescence; plasma membrane platelets; phosphatidylinositol-4,5-bisphosphate; phosphatidylinositol-4-monophosphate; immunofluorescence; plasma membrane
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MDPI and ACS Style

Bura, A.; Jurak Begonja, A. Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets. Life 2021, 11, 1331. https://doi.org/10.3390/life11121331

AMA Style

Bura A, Jurak Begonja A. Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets. Life. 2021; 11(12):1331. https://doi.org/10.3390/life11121331

Chicago/Turabian Style

Bura, Ana, and Antonija Jurak Begonja. 2021. "Imaging of Intracellular and Plasma Membrane Pools of PI(4,5)P2 and PI4P in Human Platelets" Life 11, no. 12: 1331. https://doi.org/10.3390/life11121331

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