A Role for Sphingomyelin-Rich Lipid Domains in the Accumulation of Phosphatidylinositol-4,5-Bisphosphate to the Cleavage Furrow during Cytokinesis

Lipid Biology Laboratory, RIKEN Advanced Science Institute, Wako, Saitama, Japan.
Molecular and Cellular Biology (Impact Factor: 4.78). 02/2012; 32(8):1396-407. DOI: 10.1128/MCB.06113-11
Source: PubMed


Cytokinesis is a crucial step in the creation of two daughter cells by the formation and ingression of the cleavage furrow. Here, we show that sphingomyelin (SM), one of the major sphingolipids in mammalian cells, is required for the localization of phosphatidylinositol-4,5-bisphosphate (PIP(2)) to the cleavage furrow during cytokinesis. Real-time observation with a labeled SM-specific protein, lysenin, revealed that SM is concentrated in the outer leaflet of the furrow at the time of cytokinesis. Superresolution fluorescence microscopy analysis indicates a transbilayer colocalization between the SM-rich domains in the outer leaflet and PIP(2)-rich domains in the inner leaflet of the plasma membrane. The depletion of SM disperses PIP(2) and inhibits the recruitment of the small GTPase RhoA to the cleavage furrow, leading to abnormal cytokinesis. These results suggest that the formation of SM-rich domains is required for the accumulation of PIP(2) to the cleavage furrow, which is a prerequisite for the proper translocation of RhoA and the progression of cytokinesis.

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    • "Measuring interactions: co-localization and clustering Already relatively early in the development of localization microscopy, different techniques were developed to quantify clustering/grouping of the probe. Popular techniques are Ripley's K, H and L statistics (Abe et al. 2012; Hess et al. 2007; Lee et al. 2011; Lehmann et al. 2011; Owen et al. 2010) and pair correlation (Notelaers et al. 2014; Sengupta et al. 2011, 2013; Shelby et al. 2013; Sherman et al. 2013; Veatch et al. 2012), which deliver clustering statistics in a single step starting from the set of localized positions. Other techniques require a two-step process in which individual clusters are first identified and then analyzed in a second step (Bar-On et al. 2012; Buss et al. 2013; Greenfield et al. 2009; Itano et al. 2012; Kaufmann et al. 2011; Manley et al. 2008; Nan et al. 2013; Nickerson et al. 2014; Ori et al. 2013; Rossy et al. 2013; Scarselli et al. 2012; Sherman et al. 2011; Wang et al. 2014a; Williamson et al. 2011). "
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    Cell and Tissue Research 02/2015; 360(1). DOI:10.1007/s00441-014-2109-0 · 3.57 Impact Factor
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    • "They detected aligned PI(4,5)P 2 /SM clusters that were on average ~250 nm [26]. SM clustering appeared to be required for PI(4,5)P 2 domain formation [26]. These studies provide evidence that cytoplasmic leaflet PI(4,5)P 2 domains may correspond in part to cholesterol/SM-rich raft domains. "
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    Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids 09/2014; 1851(6). DOI:10.1016/j.bbalip.2014.09.017 · 5.16 Impact Factor
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    • "SM content affects PI(4,5)P2-PLCγ signaling in synaptosomes Alterations in phosphoinositide PI(4,5)P2 levels seem to be at the base of neuronal dysfunction in different cognitive pathological conditions and in aging (Landman et al., 2006; Berman et al., 2008). In turn, phosphoinositide clustering is critically dependent on SM-rich domains (Abe et al., 2012). These two series of evidence led us to propose that the cognitive deficits in ASMko mice may be due, at least in part, to SMmediated PI(4,5)P2 signaling defects. "
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    ABSTRACT: Niemann-Pick disease type A (NPDA) is a fatal disease due to mutations in the acid sphingomyelinase (ASM) gene, which triggers the abnormal accumulation of sphingomyelin (SM) in lysosomes and the plasma membrane of mutant cells. Although the disease affects multiple organs, the impact on the brain is the most invalidating feature. The mechanisms responsible for the cognitive deficits characteristic of this condition are only partially understood. Using mice lacking the ASM gene (ASMKO), a model system in NPDA research, we here report that high sphingomyelin levels in mutant neurons lead to low synaptic levels of phosphoinositide PI(4,5)P2 and reduced activity of its hydrolyzing phosphatase PLCγ, which are key players in synaptic plasticity events. In addition, mutant neurons have reduced levels of membrane-bound MARCKS, a protein required for PI(4,5)P2 membrane clustering and hydrolysis. Intracerebroventricular infusion of a peptide that mimics the effector domain of MARCKS increases the content of PI(4,5)P2 in the synaptic membrane and ameliorates behavioral abnormalities in ASMko mice.
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Mitsuhiro Abe