BODIPY-cholesterol: a new tool to visualize sterol trafficking in living cells and organisms.
Analysis of sterol distribution and transport in living cells has been hampered by the lack of bright, photostable fluorescent sterol derivatives that closely resemble cholesterol. In this study, we employed atomistic simulations and experiments to characterize a cholesterol compound with fluorescent boron dipyrromethene difluoride linked to sterol carbon-24 (BODIPY-cholesterol). This probe packed in the membrane and behaved similarly to cholesterol both in normal and in cholesterol-storage disease cells and with trace amounts allowed the visualization of sterol movement in living systems. Upon injection into the yolk sac, BODIPY-cholesterol did not disturb zebrafish development and was targeted to sterol-enriched brain regions in live fish. We conclude that this new probe closely mimics the membrane partitioning and trafficking of cholesterol and, because of its excellent fluorescent properties, enables the direct monitoring of sterol movement by time-lapse imaging using trace amounts of the probe. This is, to our knowledge, the first cholesterol probe that fulfills these prerequisites.
Hölttä-Vuori
Maarit
M
Institute of Biomedicine/Anatomy, Haartmaninkatu 8, University of Helsinki, Helsinki 00014, Finland.
Uronen
Riikka-Liisa
RL
Repakova
Jarmila
J
Salonen
Emppu
E
Vattulainen
Ilpo
I
Panula
Pertti
P
Li
Zaiguo
Z
Bittman
Robert
R
Ikonen
Elina
E
D000818:Animals; D001692:Biological Transport; D001896:Boron Compounds / Q000737:chemistry*; D016466:CHO Cells; D002784:Cholesterol / Q000737:chemistry*; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes; D007150:Immunohistochemistry; D013261:Sterols / Q000378:metabolism*; D015027:Zebrafish
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D013261:Sterols; D002784:Cholesterol