Calcium ions trigger the exposure of phosphatidylserine on the surface of necrotic cells

English version

Autoři: Yoshitaka Furuta ^aff001; Omar Pena-Ramos ^aff001; Zao Li ^aff001; Lucia Chiao ^aff001; Zheng Zhou ^aff001
Působiště autorů: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, United States of America ^aff001; School of Pharmacy, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, Japan ^aff002
Vyšlo v časopise: Calcium ions trigger the exposure of phosphatidylserine on the surface of necrotic cells. PLoS Genet 17(2): e1009066. doi:10.1371/journal.pgen.1009066
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1009066

Souhrn

Intracellular Ca²⁺ level is under strict regulation through calcium channels and storage pools including the endoplasmic reticulum (ER). Mutations in certain ion channel subunits, which cause mis-regulated Ca²⁺ influx, induce the excitotoxic necrosis of neurons. In the nematode Caenorhabditis elegans, dominant mutations in the DEG/ENaC sodium channel subunit MEC-4 induce six mechanosensory (touch) neurons to undergo excitotoxic necrosis. These necrotic neurons are subsequently engulfed and digested by neighboring hypodermal cells. We previously reported that necrotic touch neurons actively expose phosphatidylserine (PS), an “eat-me” signal, to attract engulfing cells. However, the upstream signal that triggers PS externalization remained elusive. Here we report that a robust and transient increase of cytoplasmic Ca²⁺ level occurs prior to the exposure of PS on necrotic touch neurons. Inhibiting the release of Ca²⁺ from the ER, either pharmacologically or genetically, specifically impairs PS exposure on necrotic but not apoptotic cells. On the contrary, inhibiting the reuptake of cytoplasmic Ca²⁺ into the ER induces ectopic necrosis and PS exposure. Remarkably, PS exposure occurs independently of other necrosis events. Furthermore, unlike in mutants of DEG/ENaC channels, in dominant mutants of deg-3 and trp-4, which encode Ca²⁺ channels, PS exposure on necrotic neurons does not rely on the ER Ca²⁺ pool. Our findings indicate that high levels of cytoplasmic Ca²⁺ are necessary and sufficient for PS exposure. They further reveal two Ca²⁺-dependent, necrosis-specific pathways that promote PS exposure, a “two-step” pathway initiated by a modest influx of Ca²⁺ and further boosted by the release of Ca²⁺ from the ER, and another, ER-independent, pathway. Moreover, we found that ANOH-1, the worm homolog of mammalian phospholipid scramblase TMEM16F, is necessary for efficient PS exposure in thapsgargin-treated worms and trp-4 mutants, like in mec-4 mutants. We propose that both the ER-mediated and ER-independent Ca²⁺ pathways promote PS externalization through activating ANOH-1.

Klíčová slova:

Apoptosis – Caenorhabditis elegans – Cytoplasm – Fluorescence imaging – Larvae – Motor neurons – Necrosis – Neurons

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