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PMID: 8188746 Published · ppublish English Journal Article

The kinetic aspects of intracellular fluorescence labeling with TMA-DPH support the maturation model for endocytosis in L929 cells.

The Journal of cell biology ·Vol. 125 ·No. 4 ·1994-05-00 ·Pages 783-94

Illinger D, Kuhry JG

Abstract

TMA-DPH (1-(4-trimethylammonium)-6-phenyl-1,3,5-hexatriene), a hydrophobic fluorescent membrane probe, interacts with living cells by instantaneous incorporation into the plasma membrane, where it becomes fluorescent. It then follows the intracellular constitutive membrane traffic and acts as a bulk membrane marker of the endocytic pathway (Illinger, D., P. Poindron, P. Fonteneau, M. Modolell, and J. G. Kuhry. 1990. Biochim. Biophys. Acta. 1030:73-81; Illinger, D., P. Poindron, and J. G. Kuhry. 1991. Biol. Cell. 73:131-138). As such, TMA-DPH displays particular properties mainly due to partition between membranes and aqueous media. From these properties, original arguments can be inferred in favor of the maturation model for the endocytic pathway, against that of pre-existing compartments, in L929 cultured mouse fibroblasts. (a) TMA-DPH labeling is seen to progress from the cell periphery to perinuclear regions during endocytosis without any noticeable loss in fluorescence intensity; with a vesicle shuttle model this evolution would be accompanied by probe dilution with a decrease in the overall intracellular fluorescence intensity, and the labeling of the inner (late) compartments could in no way become more intense than that of the peripheral (early) ones. (b) From TMA-DPH fluorescence anisotropy assays, it is concluded that membrane fluidity is the same in the successive endocytic compartments as in the plasma membrane, which probably denotes a similar phospholipidic membrane composition, as might be expected in the maturation model. (c) TMA-DPH internalization and release kinetics are more easily described with the maturation model.

MeSH Terms
Animals Cell Line Diphenylhexatriene/analogs & derivatives,metabolism Endocytosis/physiology Fluorescence Polarization Fluorescent Dyes/metabolism Kinetics Mice Models, Biological
Chemicals
Fluorescent Dyes Diphenylhexatriene 1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Illinger D
Laboratoire de Biophysique, URA491 du Centre National de la Recherche Scientifique, Université Louis Pasteur, Strasbourg, France.
Kuhry J G
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.125.4.783
Published
1994-05-00
Pages
783-94
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120073
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.125.4.783 PMC Full Text View journal details
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