Surfactant uptake dynamics in mammalian cells elucidated with quantitative coherent anti-stokes Raman scattering microspectroscopy.
The mechanism of surfactant-induced cell lysis has been studied with quantitative coherent anti-Stokes Raman scattering (CARS) microspectroscopy. The dynamics of surfactant molecules as well as intracellular biomolecules in living Chinese Hamster Lung (CHL) cells has been examined for a low surfactant concentration (0.01 w%). By using an isotope labeled surfactant having CD bonds, surfactant uptake dynamics in living cells has been traced in detail. The simultaneous CARS imaging of the cell itself and the internalized surfactant has shown that the surfactant molecules is first accumulated inside a CHL cell followed by a sudden leak of cytosolic components such as proteins to the outside of the cell. This finding indicates that surfactant uptake occurs prior to the cell lysis, contrary to what has been believed: surface adsorption of surfactant molecules has been thought to occur first with subsequent disruption of cell membranes. Quantitative CARS microspectroscopy enables us to determine the molecular concentration of the surfactant molecules accumulated in a cell. We have also investigated the effect of a drug, nocodazole, on the surfactant uptake dynamics. As a result of the inhibition of tubulin polymerization by nocodazole, the surfactant uptake rate is significantly lowered. This fact suggests that intracellular membrane trafficking contributes to the surfactant uptake mechanism.
Department of Chemistry, School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Masanari
M
Okuno
Department of Chemistry, School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Hideaki
H
Kano
Safety Science Research Laboratories, Kao Corporation, Haga-Gun, Tochigi, Japan.
Kenkichi
K
Fujii
Analytical Science Research Laboratories, Kao Corporation, Haga-Gun, Tochigi, Japan.
Kotatsu
K
Bito
Analytical Science Research Laboratories, Kao Corporation, Haga-Gun, Tochigi, Japan.
Satoru
S
Naito
Institut de Recherche XLIM, UMR CNRS, Limoges, France.
Philippe
P
Leproux
Institut de Recherche XLIM, UMR CNRS, Limoges, France.
Vincent
V
Couderc
Department of Chemistry, School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan; Institute of Molecular Science and Department of Applied Chemistry, National Chiao Tung University, Hsinchu, Taiwan.
Hiro-o
HO
Hamaguchi
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D001693:Biological Transport, Active / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D015739:Nocodazole / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D013059:Spectrum Analysis, Raman; D013501:Surface-Active Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D014404:Tubulin / Q000378:metabolism
D000970:Antineoplastic Agents; D013501:Surface-Active Agents; D014404:Tubulin; D015739:Nocodazole