Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADPH oxidase activity in neutrophils

Jens Christian Brasen; Lars Folke Olsen; Maurice B. Hallett

doi:10.1016/j.bbamcr.2011.05.002

Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADPH oxidase activity in neutrophils

Jens Christian Brasen, Lars Folke Olsen, Maurice B. Hallett

Research output: Contribution to journal › Journal article › Research › peer-review

Abstract

In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast damped Ca2+ spikes with a period of 15 s and slower irregular spikes with a period greater than 50 s. Spikes in Ca2+ occurred in the absence of Ca2+ influx, but the amplitude was damped by inhibition of Ca2+ influx. Using the oxidation of hydroethidine as a cytosolic marker of oxidant production, we show that the generation of reactive oxygen species by neutrophils adherent to glass was accelerated by ATP. The step-up in NADPH oxidase activity followed the first elevation of cytosolic Ca2+ but, despite subsequent spikes in Ca2+ concentration, no oscillations in oxidase activity could be detected. ATP induced spikes in Ca2+ in a very reproducible way and we propose that the Ca2+ signal is an on-switch for oxidase activity, but the activity is apparently not directly correlated with spiking activity in cytosolic Ca2+. (C) 2011 Elsevier B.V. All rights reserved.

Original language	English
Journal	B B A - Molecular Cell Research
Volume	1813
Issue number	8
Pages (from-to)	1446-1452
ISSN	0167-4889
DOIs	https://doi.org/10.1016/j.bbamcr.2011.05.002
Publication status	Published - 2011
Externally published	Yes

Keywords

ATP
Calcium
Ca2+
Spikes
Neutrophils
NADPH oxidase
BIOCHEMISTRY
CELL
ADHERENT HUMAN-NEUTROPHILS
PROTEIN-KINASE-C
INTRACELLULAR SUPEROXIDE
PLASMA-MEMBRANE
CALCIUM INFLUX
CA-2+ INFLUX
OSCILLATIONS
HYDROETHIDINE
PHAGOCYTOSIS
FLUORESCENCE
calcium concentration
enzyme activity
spiking activity
Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common
ATP 111839-44-2
calcium ion 14127-61-8
glass
hydroethidine 104821-25-2 oxidation
NADPH oxidase complex
reactive oxygen species 7782-44-7
02506, Cytology - Animal
02508, Cytology - Human
10060, Biochemistry studies - General
10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
10069, Biochemistry studies - Minerals
10802, Enzymes - General and comparative studies: coenzymes
15002, Blood - Blood and lymph studies
15004, Blood - Blood cell studies
34502, Immunology - General and methods
Biochemistry and Molecular Biophysics
Chemical Coordination and Homeostasis
cytosol
neutrophil immune system, blood and lymphatics
confocal microscopy laboratory techniques, imaging and microscopy techniques
Enzymology
Immune System

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10.1016/j.bbamcr.2011.05.002

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title = "Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADPH oxidase activity in neutrophils",

abstract = "In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast damped Ca2+ spikes with a period of 15 s and slower irregular spikes with a period greater than 50 s. Spikes in Ca2+ occurred in the absence of Ca2+ influx, but the amplitude was damped by inhibition of Ca2+ influx. Using the oxidation of hydroethidine as a cytosolic marker of oxidant production, we show that the generation of reactive oxygen species by neutrophils adherent to glass was accelerated by ATP. The step-up in NADPH oxidase activity followed the first elevation of cytosolic Ca2+ but, despite subsequent spikes in Ca2+ concentration, no oscillations in oxidase activity could be detected. ATP induced spikes in Ca2+ in a very reproducible way and we propose that the Ca2+ signal is an on-switch for oxidase activity, but the activity is apparently not directly correlated with spiking activity in cytosolic Ca2+. (C) 2011 Elsevier B.V. All rights reserved.",

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author = "Brasen, {Jens Christian} and Olsen, {Lars Folke} and Hallett, {Maurice B.}",

year = "2011",

doi = "10.1016/j.bbamcr.2011.05.002",

language = "English",

volume = "1813",

pages = "1446--1452",

journal = "B B A - Molecular Cell Research",

issn = "0167-4889",

publisher = "Elsevier",

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TY - JOUR

T1 - Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADPH oxidase activity in neutrophils

AU - Brasen, Jens Christian

AU - Olsen, Lars Folke

AU - Hallett, Maurice B.

PY - 2011

Y1 - 2011

N2 - In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast damped Ca2+ spikes with a period of 15 s and slower irregular spikes with a period greater than 50 s. Spikes in Ca2+ occurred in the absence of Ca2+ influx, but the amplitude was damped by inhibition of Ca2+ influx. Using the oxidation of hydroethidine as a cytosolic marker of oxidant production, we show that the generation of reactive oxygen species by neutrophils adherent to glass was accelerated by ATP. The step-up in NADPH oxidase activity followed the first elevation of cytosolic Ca2+ but, despite subsequent spikes in Ca2+ concentration, no oscillations in oxidase activity could be detected. ATP induced spikes in Ca2+ in a very reproducible way and we propose that the Ca2+ signal is an on-switch for oxidase activity, but the activity is apparently not directly correlated with spiking activity in cytosolic Ca2+. (C) 2011 Elsevier B.V. All rights reserved.

AB - In order to establish whether non-mitochondrial oxidase activity in human neutrophils is tightly related to cytosolic Ca2+ concentration, we simultaneously measured Ca2+ oscillations induced by ATP and oxidant production in single adherent neutrophils using confocal microscopy. ATP induced fast damped Ca2+ spikes with a period of 15 s and slower irregular spikes with a period greater than 50 s. Spikes in Ca2+ occurred in the absence of Ca2+ influx, but the amplitude was damped by inhibition of Ca2+ influx. Using the oxidation of hydroethidine as a cytosolic marker of oxidant production, we show that the generation of reactive oxygen species by neutrophils adherent to glass was accelerated by ATP. The step-up in NADPH oxidase activity followed the first elevation of cytosolic Ca2+ but, despite subsequent spikes in Ca2+ concentration, no oscillations in oxidase activity could be detected. ATP induced spikes in Ca2+ in a very reproducible way and we propose that the Ca2+ signal is an on-switch for oxidase activity, but the activity is apparently not directly correlated with spiking activity in cytosolic Ca2+. (C) 2011 Elsevier B.V. All rights reserved.

KW - ATP

KW - Calcium

KW - Ca2+

KW - Spikes

KW - Neutrophils

KW - NADPH oxidase

KW - BIOCHEMISTRY

KW - CELL

KW - ADHERENT HUMAN-NEUTROPHILS

KW - PROTEIN-KINASE-C

KW - INTRACELLULAR SUPEROXIDE

KW - PLASMA-MEMBRANE

KW - CALCIUM INFLUX

KW - CA-2+ INFLUX

KW - OSCILLATIONS

KW - HYDROETHIDINE

KW - PHAGOCYTOSIS

KW - FLUORESCENCE

KW - calcium concentration

KW - enzyme activity

KW - spiking activity

KW - Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common

KW - ATP 111839-44-2

KW - calcium ion 14127-61-8

KW - glass

KW - hydroethidine 104821-25-2 oxidation

KW - NADPH oxidase complex

KW - reactive oxygen species 7782-44-7

KW - 02506, Cytology - Animal

KW - 02508, Cytology - Human

KW - 10060, Biochemistry studies - General

KW - 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines

KW - 10069, Biochemistry studies - Minerals

KW - 10802, Enzymes - General and comparative studies: coenzymes

KW - 15002, Blood - Blood and lymph studies

KW - 15004, Blood - Blood cell studies

KW - 34502, Immunology - General and methods

KW - Biochemistry and Molecular Biophysics

KW - Chemical Coordination and Homeostasis

KW - cytosol

KW - neutrophil immune system, blood and lymphatics

KW - confocal microscopy laboratory techniques, imaging and microscopy techniques

KW - Enzymology

KW - Immune System

U2 - 10.1016/j.bbamcr.2011.05.002

DO - 10.1016/j.bbamcr.2011.05.002

M3 - Journal article

C2 - 21596069

SN - 0167-4889

VL - 1813

SP - 1446

EP - 1452

JO - B B A - Molecular Cell Research

JF - B B A - Molecular Cell Research

IS - 8

ER -

Extracellular ATP induces spikes in cytosolic free Ca2+ but not in NADPH oxidase activity in neutrophils

Abstract

Keywords

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