Fluorescent lipid probes 12-AS and TMA-DPH report on selective, purinergically induced fluidity changes in plasma membranes of lymphoid cells

J. Matkó, Péter Nagy

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8 Citations (Scopus)

Abstract

The effect of extracellular ATP (ATP(ex)) on the anisotropy of 1-[4-(trimethylamino) phenyl]-6-phenyl-hexa-3,5 triene (TMA-DPH) and 12-anthroyloxi-stearic acid (12-AS) fluorescence was investigated in Balb/C mouse thymocytes and in JY human lymphoblasts. These cells have been shown recently to be sensitive and resistant to ATP(ex), respectively, in terms of cellular responses. Extracellular ATP (1 mM) induced a time-dependent elevation in the emission anisotropy of both probes (indicating an increased lipid packing density) in the plasma membrane of thymocytes. The maximal effect, at 37°C, was observed between 20 and 60 min after ATP(ex) administration, and followed by a gradual decrease of fluorescence anisotropy at longer times (60-180 min). ATP(ex) did not change membrane fluidity of thymocytes below the phase transition temperature (at 18°C). Oxidized ATP (oATP), a selective antagonist of P2, purinoreceptors, blocked the ATP(ex)-induced decrease in membrane fluidity. Low ATP(ex) concentrations (100-300 μM) - which are known to induce distinct signals (changes in membrane potential and intracellular Ph) - slightly fluidized the plasma membrane of thymocytes. This effect was partially blocked by quinine, a blocker of Ca2+ - activated K+ channels. Neither 12-AS nor TMA-DPH showed any change in their emission anisotropy upon ATP(ex) - treatment in the plasma membrane of the resistant human JY lymphoblast cells. No other signalling event (membrane potential change, Ca2+ response) is elicited by ATP(ex) in this cell line. These data suggest that the changes in the membrane fluidity are likely consequences of specific, purinoreceptor-mediated signalling events, such as hyper-or depolarization of the plasma membrane or Ca2+ influx. These signals may induce changes in the conformation or lateral organization of membrane proteins, perturbing protein-lipid interactions, as well.

Original languageEnglish
Pages (from-to)120-125
Number of pages6
JournalJournal of Photochemistry and Photobiology, B: Biology
Volume40
Issue number2
DOIs
Publication statusPublished - Sep 1997

Fingerprint

Stearic acid
thymocytes
Fluidity
adenosine triphosphate
Adenosinetriphosphate
Cell membranes
stearic acid
Fluorescent Dyes
Lipids
membrane fluidity
lipids
plasma membrane
Adenosine Triphosphate
Cell Membrane
Lymphocytes
membranes
acids
probes
cells
membrane potential

Keywords

  • Cell death
  • Extracellular ATP
  • Fluidity
  • Membrane pores
  • Purinoreceptors
  • Signalling

ASJC Scopus subject areas

  • Plant Science
  • Bioengineering
  • Physical and Theoretical Chemistry

Cite this

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title = "Fluorescent lipid probes 12-AS and TMA-DPH report on selective, purinergically induced fluidity changes in plasma membranes of lymphoid cells",
abstract = "The effect of extracellular ATP (ATP(ex)) on the anisotropy of 1-[4-(trimethylamino) phenyl]-6-phenyl-hexa-3,5 triene (TMA-DPH) and 12-anthroyloxi-stearic acid (12-AS) fluorescence was investigated in Balb/C mouse thymocytes and in JY human lymphoblasts. These cells have been shown recently to be sensitive and resistant to ATP(ex), respectively, in terms of cellular responses. Extracellular ATP (1 mM) induced a time-dependent elevation in the emission anisotropy of both probes (indicating an increased lipid packing density) in the plasma membrane of thymocytes. The maximal effect, at 37°C, was observed between 20 and 60 min after ATP(ex) administration, and followed by a gradual decrease of fluorescence anisotropy at longer times (60-180 min). ATP(ex) did not change membrane fluidity of thymocytes below the phase transition temperature (at 18°C). Oxidized ATP (oATP), a selective antagonist of P2, purinoreceptors, blocked the ATP(ex)-induced decrease in membrane fluidity. Low ATP(ex) concentrations (100-300 μM) - which are known to induce distinct signals (changes in membrane potential and intracellular Ph) - slightly fluidized the plasma membrane of thymocytes. This effect was partially blocked by quinine, a blocker of Ca2+ - activated K+ channels. Neither 12-AS nor TMA-DPH showed any change in their emission anisotropy upon ATP(ex) - treatment in the plasma membrane of the resistant human JY lymphoblast cells. No other signalling event (membrane potential change, Ca2+ response) is elicited by ATP(ex) in this cell line. These data suggest that the changes in the membrane fluidity are likely consequences of specific, purinoreceptor-mediated signalling events, such as hyper-or depolarization of the plasma membrane or Ca2+ influx. These signals may induce changes in the conformation or lateral organization of membrane proteins, perturbing protein-lipid interactions, as well.",
keywords = "Cell death, Extracellular ATP, Fluidity, Membrane pores, Purinoreceptors, Signalling",
author = "J. Matk{\'o} and P{\'e}ter Nagy",
year = "1997",
month = "9",
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volume = "40",
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journal = "Journal of Photochemistry and Photobiology B: Biology",
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TY - JOUR

T1 - Fluorescent lipid probes 12-AS and TMA-DPH report on selective, purinergically induced fluidity changes in plasma membranes of lymphoid cells

AU - Matkó, J.

AU - Nagy, Péter

PY - 1997/9

Y1 - 1997/9

N2 - The effect of extracellular ATP (ATP(ex)) on the anisotropy of 1-[4-(trimethylamino) phenyl]-6-phenyl-hexa-3,5 triene (TMA-DPH) and 12-anthroyloxi-stearic acid (12-AS) fluorescence was investigated in Balb/C mouse thymocytes and in JY human lymphoblasts. These cells have been shown recently to be sensitive and resistant to ATP(ex), respectively, in terms of cellular responses. Extracellular ATP (1 mM) induced a time-dependent elevation in the emission anisotropy of both probes (indicating an increased lipid packing density) in the plasma membrane of thymocytes. The maximal effect, at 37°C, was observed between 20 and 60 min after ATP(ex) administration, and followed by a gradual decrease of fluorescence anisotropy at longer times (60-180 min). ATP(ex) did not change membrane fluidity of thymocytes below the phase transition temperature (at 18°C). Oxidized ATP (oATP), a selective antagonist of P2, purinoreceptors, blocked the ATP(ex)-induced decrease in membrane fluidity. Low ATP(ex) concentrations (100-300 μM) - which are known to induce distinct signals (changes in membrane potential and intracellular Ph) - slightly fluidized the plasma membrane of thymocytes. This effect was partially blocked by quinine, a blocker of Ca2+ - activated K+ channels. Neither 12-AS nor TMA-DPH showed any change in their emission anisotropy upon ATP(ex) - treatment in the plasma membrane of the resistant human JY lymphoblast cells. No other signalling event (membrane potential change, Ca2+ response) is elicited by ATP(ex) in this cell line. These data suggest that the changes in the membrane fluidity are likely consequences of specific, purinoreceptor-mediated signalling events, such as hyper-or depolarization of the plasma membrane or Ca2+ influx. These signals may induce changes in the conformation or lateral organization of membrane proteins, perturbing protein-lipid interactions, as well.

AB - The effect of extracellular ATP (ATP(ex)) on the anisotropy of 1-[4-(trimethylamino) phenyl]-6-phenyl-hexa-3,5 triene (TMA-DPH) and 12-anthroyloxi-stearic acid (12-AS) fluorescence was investigated in Balb/C mouse thymocytes and in JY human lymphoblasts. These cells have been shown recently to be sensitive and resistant to ATP(ex), respectively, in terms of cellular responses. Extracellular ATP (1 mM) induced a time-dependent elevation in the emission anisotropy of both probes (indicating an increased lipid packing density) in the plasma membrane of thymocytes. The maximal effect, at 37°C, was observed between 20 and 60 min after ATP(ex) administration, and followed by a gradual decrease of fluorescence anisotropy at longer times (60-180 min). ATP(ex) did not change membrane fluidity of thymocytes below the phase transition temperature (at 18°C). Oxidized ATP (oATP), a selective antagonist of P2, purinoreceptors, blocked the ATP(ex)-induced decrease in membrane fluidity. Low ATP(ex) concentrations (100-300 μM) - which are known to induce distinct signals (changes in membrane potential and intracellular Ph) - slightly fluidized the plasma membrane of thymocytes. This effect was partially blocked by quinine, a blocker of Ca2+ - activated K+ channels. Neither 12-AS nor TMA-DPH showed any change in their emission anisotropy upon ATP(ex) - treatment in the plasma membrane of the resistant human JY lymphoblast cells. No other signalling event (membrane potential change, Ca2+ response) is elicited by ATP(ex) in this cell line. These data suggest that the changes in the membrane fluidity are likely consequences of specific, purinoreceptor-mediated signalling events, such as hyper-or depolarization of the plasma membrane or Ca2+ influx. These signals may induce changes in the conformation or lateral organization of membrane proteins, perturbing protein-lipid interactions, as well.

KW - Cell death

KW - Extracellular ATP

KW - Fluidity

KW - Membrane pores

KW - Purinoreceptors

KW - Signalling

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