Membrane potential dynamics of C5a-stimulated neutrophil granulocytes.

Publisher: Springer Country of Publication: Germany NLM ID: 0154720 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-2013 (Electronic) Linking ISSN: 00316768 NLM ISO Abbreviation: Pflugers Arch Subsets: MEDLINE

Original Publication: Berlin, New York, Springer.

Neutrophils*/metabolism ; Neutrophils*/drug effects ; Neutrophils*/physiology ; Complement C5a*/metabolism ; Complement C5a*/pharmacology ; Reactive Oxygen Species*/metabolism ; Mice, Inbred C57BL* ; Membrane Potentials*/physiology; Animals ; Mice ; NADPH Oxidases/metabolism ; Actin Cytoskeleton/metabolism ; Tumor Necrosis Factor-alpha/metabolism ; Tumor Necrosis Factor-alpha/pharmacology ; Cell Movement/drug effects ; Neutrophil Activation ; NADPH Oxidase 2/metabolism

Neutrophil granulocytes play a crucial role in host defense against invading pathogens and in inflammatory diseases. The aim of this study was to elucidate membrane potential dynamics during the initial phase of neutrophil activation and its relation to migration and production of reactive oxygen species (ROS). We performed ROS production measurements of neutrophils from healthy C57BL/6J mice after TNFα-priming and/or C5a stimulation. The actin cytoskeleton was visualized with fluorescence microscopy. Furthermore, we combined migration assays and measurements of membrane potential dynamics after stimulating unprimed and/or TNFα-primed neutrophils with C5a. We show that C5a has a concentration-dependent effect on ROS production and chemokinetic migration. Chemokinetic migration and chemotaxis are impaired at C5a concentrations that induce ROS production. The actin cytoskeleton of unstimulated and of ROS-producing neutrophils is not distributed in a polarized way. Inhibition of the phagocytic NADPH oxidase NOX2 with diphenyleneiodonium (DPI) leads to a polarized distribution of the actin cytoskeleton and rescues chemokinetic migration of primed and C5a-stimulated neutrophils. Moreover, C5a evokes a pronounced depolarization of the cell membrane potential by 86.6 ± 4.2 mV starting from a resting membrane potential of -74.3 ± 0.7 mV. The C5a-induced depolarization occurs almost instantaneously (within less than one minute) in contrast to the more gradually developing depolarization induced by PMA (lag time of 3-4 min). This initial depolarization is accompanied by a decrease of the migration velocity. Collectively, our results show that stimulation with C5a evokes parallel changes in membrane potential dynamics, neutrophil ROS production and motility. Notably, the amplitude of membrane potential dynamics is comparable to that of excitable cells.
(© 2024. The Author(s).)

Aging (Albany NY). 2017 Dec 16;9(12):2610-2628. (PMID: 29253841)
Am J Pathol. 2023 Apr;193(4):474-492. (PMID: 36669683)
Pflugers Arch. 2023 Oct;475(10):1225-1240. (PMID: 37566113)
Antioxid Redox Signal. 2014 Aug 20;21(6):915-32. (PMID: 24053140)
Front Physiol. 2011 Mar 11;2:10. (PMID: 21483726)
Front Immunol. 2020 Sep 08;11:2124. (PMID: 33013896)
Biochem J. 2000 Apr 1;347 Pt 1:285-9. (PMID: 10727429)
Nat Rev Immunol. 2007 Sep;7(9):678-89. (PMID: 17717539)
J Immunol. 2003 Jul 15;171(2):893-901. (PMID: 12847259)
Redox Biol. 2021 Nov;47:102133. (PMID: 34562872)
PLoS Comput Biol. 2022 May 18;18(5):e1010089. (PMID: 35584137)
J Immunol. 2022 Jul 1;209(1):136-144. (PMID: 35715008)
Blood. 2004 Nov 1;104(9):2947-53. (PMID: 15251984)
Proc Natl Acad Sci U S A. 2020 Sep 29;117(39):24403-24414. (PMID: 32929002)
Int J Mol Sci. 2019 Sep 10;20(18):. (PMID: 31510052)
Immunity. 2012 Dec 14;37(6):1037-49. (PMID: 23159440)
Blood Cells Mol Dis. 2012 Jan 15;48(1):53-61. (PMID: 22134107)
Immunol Rev. 2016 Sep;273(1):194-218. (PMID: 27558336)
Am J Physiol Cell Physiol. 2015 Feb 1;308(3):C264-76. (PMID: 25472961)
Biochimie. 2007 Sep;89(9):1089-106. (PMID: 17428601)
Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6885-9. (PMID: 10823889)
Nat Rev Immunol. 2013 Mar;13(3):159-75. (PMID: 23435331)
Biochem J. 1988 Oct 1;255(1):285-90. (PMID: 2848506)
J Exp Med. 2010 Jan 18;207(1):129-39. (PMID: 20026664)
Plant J. 2008 Jul;55(1):161-73. (PMID: 18363788)
J Leukoc Biol. 2020 Feb;107(2):285-297. (PMID: 31841231)
Biochem Biophys Res Commun. 2019 Jun 4;513(3):708-713. (PMID: 30987827)
Cells. 2022 Sep 16;11(18):. (PMID: 36139464)
J Biol Chem. 1980 Oct 10;255(19):9494-500. (PMID: 6251091)
Cytometry A. 2013 Jul;83(7):612-26. (PMID: 23650268)
Eur J Clin Invest. 2018 Nov;48 Suppl 2:e12964. (PMID: 29873837)
Eur J Immunol. 2015 Jul;45(7):2143-53. (PMID: 25912155)
Cardiovasc Res. 2022 Mar 25;118(5):1289-1302. (PMID: 33881519)
J Biol Chem. 1999 Sep 10;274(37):26098-104. (PMID: 10473559)
J Leukoc Biol. 2018 Jan;103(1):87-98. (PMID: 28798145)
J Immunol. 2003 Mar 1;170(5):2670-9. (PMID: 12594296)
Nature. 2003 Apr 3;422(6931):531-4. (PMID: 12673252)
Acta Physiol (Oxf). 2016 Jan;216(1):132-45. (PMID: 26138196)
Redox Biol. 2015 Dec;6:260-271. (PMID: 26296072)
J Immunol. 2013 Jun 1;190(11):5496-505. (PMID: 23636057)
Nature. 1998 Apr 16;392(6677):734-7. (PMID: 9565037)
Physiol Rev. 1999 Jul;79(3):763-854. (PMID: 10390518)
Immunol Rev. 2016 Sep;273(1):180-93. (PMID: 27558335)
J Exp Med. 2006 Aug 7;203(8):1927-37. (PMID: 16880254)
Biophys J. 1993 Oct;65(4):1590-8. (PMID: 7506066)

GRK 2515/1 Deutsche Forschungsgemeinschaft; SCHW 407/17-1 Deutsche Forschungsgemeinschaft; SH 167/9-1 Deutsche Forschungsgemeinschaft; Schw2/020/18 Interdisziplinäres Zentrum für Klinische Forschung, Universitätsklinikum Münster

Keywords: Chemotaxis; Membrane potential dynamics; Neutrophils; ROS

80295-54-1 (Complement C5a)
0 (Reactive Oxygen Species)
EC 1.6.3.- (NADPH Oxidases)
0 (Tumor Necrosis Factor-alpha)
EC 1.6.3.- (NADPH Oxidase 2)

Date Created: 20240413 Date Completed: 20240530 Latest Revision: 20240920

20240921

PMC11139730

10.1007/s00424-024-02947-8

38613695