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Stages of NETosis Development upon Stimulation of Neutrophils with Activators of Different Types.
Inozemtsev, Vladimir; Sergunova, Viktoria; Vorobjeva, Nina; Kozlova, Elena; Sherstyukova, Ekaterina; Lyapunova, Snezhanna; Chernysh, Aleksandr.
Afiliação
  • Inozemtsev V; Laboratory of Biophysics of Cell Membranes under Critical State, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, V.A. Negovsky Scientific Research Institute of General Reanimatology, Petrovka Street 25c2, 107031 Moscow, Russia.
  • Sergunova V; Koltsov Institute of Developmental Biology, Russian Academy of Sciences, 26 Vavilov Street, 119334 Moscow, Russia.
  • Vorobjeva N; Laboratory of Biophysics of Cell Membranes under Critical State, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, V.A. Negovsky Scientific Research Institute of General Reanimatology, Petrovka Street 25c2, 107031 Moscow, Russia.
  • Kozlova E; Department of Immunology, Faculty of Biology, Lomonosov Moscow State University, Lenin Hills 1/12, 119234 Moscow, Russia.
  • Sherstyukova E; Laboratory of Biophysics of Cell Membranes under Critical State, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, V.A. Negovsky Scientific Research Institute of General Reanimatology, Petrovka Street 25c2, 107031 Moscow, Russia.
  • Lyapunova S; Department of Medical and Biological Physics, Sechenov First Moscow State Medical University, 119991 Moscow, Russia.
  • Chernysh A; Laboratory of Biophysics of Cell Membranes under Critical State, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, V.A. Negovsky Scientific Research Institute of General Reanimatology, Petrovka Street 25c2, 107031 Moscow, Russia.
Int J Mol Sci ; 24(15)2023 Aug 02.
Article em En | MEDLINE | ID: mdl-37569729
ABSTRACT
Before NETs are released, the neutrophil undergoes structural changes. First, it flattens, accompanied by a change in cell shape and rearrangement of the cytoskeleton. Then, nuclear swelling begins, which ends with the ejection of NETs into the extracellular space. We used widefield and confocal fluorescence microscopy to register morphological and structural changes in neutrophils during activation and NETosis. Different types of activators were used, such as NOX-dependent PMA and calcium ionophore A23187. The measurements were performed in a series of sequential stages. In the first stage (30 s after addition of activators and immediately after stimulation of neutrophils), the response of neutrophils to A23187 and PMA exposure was studied. Subsequently, the characteristics of neutrophils in different phases of activation were examined over a longer period of time (30, 60, 120, 180, and 240 min). The specific features of NETosis development were analyzed separately. During the first 30 s, neutrophils appeared to be heterogeneous in shape and structure of the actin cytoskeleton. Characteristic cell shapes included 30″ type 1 cells, similar in shape to the control, with F-actin concentrated in the center of the cytoplasm, and 30″ type 2 cells, which had flattened (spread) shapes with increased frontal dimensions and F-actin distributed throughout the cell. Later, the development of nuclear swelling, the corresponding changes in neutrophil membranes, and NET release into the extracellular space were evaluated. The conditions determining the initiation of chromatin ejection and two characteristic types of decondensed chromatin ejection were revealed. The results obtained contribute to a better understanding of the biophysical mechanisms of neutrophil activation and NETosis development.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Armadilhas Extracelulares / Neutrófilos Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Armadilhas Extracelulares / Neutrófilos Idioma: En Ano de publicação: 2023 Tipo de documento: Article