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Effects of buffering intracellular free calcium on neutrophil migration through three‐dimensional matrices
Author(s) -
Mandeville John T. H.,
Maxfield Frederick R.
Publication year - 1997
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199705)171:2<168::aid-jcp7>3.0.co;2-m
Subject(s) - vitronectin , fibronectin , integrin , microbiology and biotechnology , extracellular matrix , biophysics , adhesion , chemistry , matrix (chemical analysis) , cell migration , confocal microscopy , intracellular , cell adhesion , receptor , cell , biology , biochemistry , organic chemistry , chromatography
Repeated transient increases in intracellular free calcium levels ([Ca 2+ ] i ) are required for polymorphonuclear neutrophil migration on two‐dimensional surfaces coated with fibronectin or vitronectin. Cells in which [Ca 2+ ] i is buffered with quin2 become stuck on these substrates. Neutrophils migrating through the extracellular matrix in vivo encounter these and other substrates in a three‐dimensional architecture that may alter the spatial distribution of adhesion receptors in contact with the matrix. In this study, we used fluorescence confocal microscopy to obtain moving three‐dimensional images of neutrophils migrating through a biological tissue (human amnion) in the presence and absence of [Ca 2+ ] i ‐buffering with quin2. In the absence of buffering, [Ca 2+ ] i transients similar to those seen in cells migrating in two‐dimensions were observed. [Ca 2+ ] i ‐buffered neutrophils were able to migrate into the matrix, but they became attached firmly to the substrate at the rear of the cell, resulting in a drastically elongated morphology. Immunofluorescence revealed that neutrophils adhered to regions of the matrix that contained fibronectin. RGD‐containing peptides and antibodies that block integrin adhesion receptors for fibronectin and vitronectin were able to rescue the migration of quin2‐treated cells through three‐dimensional gels containing fibronectin and vitronectin. These data show that neutrophils migrating throughphysiologically relevant, three‐dimensional matrices undergo repetitive increases in [Ca 2+ ] i that are required for integrin‐mediated detachment from the matrix. J. Cell. Physiol. 171:168–178, 1997. © 1997 Wiley‐Liss, Inc.