Emphatic visualization of sphingomyelin-rich domains by inter-lipid FRET imaging using fluorescent sphingomyelins

Masanao Kinoshita, Hikaru Ano, Michio Murata, Kenta Shigetomi, Junichi Ikenouchi, Nobuaki Matsumori

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Imaging the distribution of sphingomyelin (SM) in membranes is an important issue in lipid-raft research. Recently we developed novel fluorescent SM analogs that exhibit partition and dynamic behaviors similar to native SM, and succeeded in visualizing lateral domain-segregation between SM-rich liquid-ordered (Lo) and SM-poor liquid-disordered (Ld) domains. However, because the fluorescent contrast between these two domains depends directly on their partition ratio for the fluorescent SMs, domain-separation becomes indeterminate when the distribution difference is not great enough. In this study, we propose the use of inter-lipid Förster resonance energy transfer (FRET) imaging between fluorescent SMs to enhance the contrast of the two domains in cases in which the inter-domain difference in SM distribution is inadequate for conventional monochromic imaging. Our results demonstrate that inter-lipid FRET intensity was significantly higher in the Lo domain than in the Ld domain, resulting in a clear and distinguishable contrast between the two domains even in poorly phase-separated giant unilamellar vesicles. In addition, we show that inter-lipid FRET imaging is useful for selective visualization of highly condensed assemblies and/or clusters of SM molecules in living cell membranes. Thus, the inter-lipid FRET imaging technique can selectively emphasize the SM-condensed domains in both artificial and biological membranes.

Original languageEnglish
Article number16801
JournalScientific reports
Volume7
Issue number1
DOIs
Publication statusPublished - Dec 1 2017

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lipids
energy transfer
liquids
partitions
membranes
rafts
imaging techniques
assemblies
analogs

All Science Journal Classification (ASJC) codes

  • General

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Emphatic visualization of sphingomyelin-rich domains by inter-lipid FRET imaging using fluorescent sphingomyelins. / Kinoshita, Masanao; Ano, Hikaru; Murata, Michio; Shigetomi, Kenta; Ikenouchi, Junichi; Matsumori, Nobuaki.

In: Scientific reports, Vol. 7, No. 1, 16801, 01.12.2017.

Research output: Contribution to journalArticle

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