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10th International Congress "Cell Volume Regulation: Novel Therapeutic Targets and Pharmacological Approaches"
HYPOCHLOROUS ACID EFFECTS ON VOLUME REGULATION OF MIGRATING NEUTROPHILS
Kuznetsova, T.G.1, Kulahava, T.A.2, Zholnerevich, I.I.2, Divakova, N.V.2, Nebieridze, M.I.3, and Semenkova, G.N.2
1 Department of Histology, Cytology & Embryology, Pirogov Russian National Medical Research University, Moscow, Russian Federation
2 Department of Biophysics, Physical Faculty, Belarusian State University, Minsk, Belarus
3 Department of Cerebral Blood Circulation and Metabolism, Ivane Beritashvili Center of Experimental Biomedicine, Tbilisi, Georgia
Cell volume regulatory mechanisms participate in different activities of cells including neutrophils migration and ROS generation [1, 2]. The effect of HOCl on the random migration of human neutrophils was studied.
Isolated neutrophils (untreated and HOCl-treated) were placed on glass for 20 min and fixed. The volume (V) and projection area (S) were calculated for single control (n = 130) and treated cells (n = 160) using atomic-force microscopy.
Because neutrophils movement consists of repeated cycles including the formation of lamellipodium during cell spreading and a cell retraction, we have identified three cell groups according to its area: 1) S < 75 |m2, inactive cells and hemispherical retractile cells; 2) S from 75 to 150 |im2, cells with growing lamellipodium; 3) S > 150 |im2, fully spreading cells.
The average V of control cells in group 2 increased by 30% as compared to group 1 cells and the values of cell volume varied around a constant mean of 149 |im3 (from 98 to 225 |im3). As volume changes associated with lamellipodium growth, as individual size differences of neutrophils may cause this volume fluctuation. With a further area rise above 150 |m2 the variability of volume
values became very low (average V 155 |m3, max V 220 |im3). The rapid increasing of cell area led to marked S/V ratio changes. It varied from 0.4 to 1.8.
The average V of exposed to HOCl cells from group 1 was less than control value (96 |im3). With cell spreading it increased initially by 57%. A slow gradual increase of cell volume was observed for the entire further area growth from 75 |im2 to the maximum. As a result the average V of treated cells reached to control value (group 2) and then overpassed (165 |im3, group 3). The fluctuation of volume values was greater and persisted in group 3. Because cell area dynamics was similar to the control, the average ratio S/V of treated cells was lower. Thus, HOCl alters cell volume regulation of random migrating human neutrophils.
References
1. Jakab, M., Ritter, M. Cell volume regulatory ion transport in the regulation of cell migration, 2006, Contrib. Nephrol., 152, pp. 161-180.
2. Kuchkina, N.V., Orlov, S.N., Pokudin, N.I., Chuchalina, A.G. Volume-dependent regulation of the respiratory burst of activated human neutrophils. Experientia, 1993, 49, pp. 995997.
Бюллетень сибирской медицины, 2013, том 12, № 4, с. 24-68
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