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Inhibition of the GTPase dynamin or actin depolymerisation initiates outward plasma membrane tubulation/vesiculation (cytoneme formation) in neutrophils
Author(s) -
Galkina Svetlana I.,
Fedorova Natalia V.,
Serebryakova Marina V.,
Arifulin Evgenii A.,
Stadnichuk Vladimir I.,
Gaponova Tatjana V.,
Baratova Ludmila A.,
Sud'ina Galina F.
Publication year - 2015
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1111/boc.201400063
Subject(s) - dynamin , biology , gtpase , microbiology and biotechnology , actin , biophysics , membrane , endocytosis , biochemistry , cell
Background information In a previous study, we demonstrated that human neutrophils can develop membrane tubulovesicular extensions (TVEs) that are 160–250 nm in width and several micrometres long. These extensions, or cytonemes, are capable of establishing long‐range contacts with other cells or bacteria. Cytonemes consist of membrane tubules and vesicles of a uniform diameter aligned in a row. The mechanism of membrane tubulation/vesiculation to form cytonemes remains unknown. Upon endocytosis, the GTPase dynamin and an intact actin cytoskeleton are required for endocytic vesicles scission from the plasma membrane. Results We examined the effects of dynasore (a dynamin specific inhibitor), and of cytochalasin D and latrunculin A (actin cytoskeleton disruption agents), on cytoneme formation in neutrophils. Scanning and transmission electron microscopy were used to observe cytoneme formation. High‐performance chromatography and mass spectrometry were used to estimate the protein composition of the cytonemes. In neutrophils, dynasore and cytochalasin D or latrunculin A initiated the formation of tubular cytonemes that were similar in diameter and composition. The formation of cytonemes in cells treated with cytochalasin D was accompanied by the appearance of tubular invaginations of the same diameter on the plasma membrane of neutrophils. The formation of dynasore‐ or cytochalasin D‐induced cytonemes, however, was blocked by the nitric oxide (NO) synthases inhibitor l ‐NAME, indicating that NO is involved in cytoneme development. Proteome analysis indicated that dynasore‐ or cytochalasin D‐induced cytonemes are secretory protrusions that contain neutrophil bactericides along with cytoplasmic proteins, such as glycolytic enzymes and actin cytoskeleton components. Conclusions Inhibition of dynamin with dynasore or actin depolymerisation with cytochalasin D or latrunculin A might impair the membrane fusion/fission events that are required for the separation of secretory vesicles from the plasma membrane and from each other. As a result, the secretory process extends from the cells as membrane TVEs or cytonemes. Modification of secretion gives neutrophils the possibility to communicate with other cells over distance via highly adhesive cellular secretory protrusions (cytonemes). Cytonemes deliver their membrane‐packed content exactly to the destination without dilution and without harm to the surrounding tissues.