Divalent cations, phospholipid asymmetry and osmotic swelling in electrically-induced lysis, cell fusion and giant cell formation with human erythrocytes.
We have previously reported that acidic phospholipids are exposed at the surface of human erythrocytes when the cells are subjected to electrical breakdown. It has now been shown that the prothrombinase assay, which was used previously for the determination of acidic phospholipids, is specific for phosphatidylserine under the conditions of our experiments. In the light of this finding, we have investigated and characterised factors that govern cell lysis, cell fusion, and the formation of giant cells induced by electrical breakdown with human erythrocytes in media of low ionic strength. Divalent cations (1.1 mM) protected the cells against haemolysis, in the order Mn2+ > Ca2+ > Ba2+ > Mg2+ >> Zn2+, whereas about 99% of the cells lysed immediately on breakdown in the presence of Na+ or K+ (2.1 mM), or Al3+ (0.95 mM). The lengths of pearl chains of fused erythrocytes formed was similarly greatest with Mn2+ and decreased progressively with Ba2+, Zn2+, Ca2+ and Mg2+. No cell fusion occurred with Na+, K+, or Al3+. It is suggested that interactions with phosphatidylserine, which is exposed at the cell surface by electrical breakdown, may enable Mn2+, Ba2+ and Ca2+ ions to inhibit cell lysis (via membrane resealing) and facilitate cell fusion. Following electrically-induced cell fusion, erythrocytes round-up into giant cells. It has previously been proposed that Ca2+ ions accelerate the rounding-up process. However, data are presented which show that, as with erythrocytes treated with Sendai virus, the formation of rounded, giant cells following cell fusion depends on the osmotic swelling properties of permeabilised erythrocytes. Osmotic swelling may also have induced any hemi-fused cells present to fuse completely. Zn2+ ions anomalously enabled erythrocytes to round-up very rapidly into giant cells following electrical breakdown. This phenomenon may result from an interaction of Zn2+ ions with cysteine groups in membrane proteins, which decreases the immediate loss of ions that occurs when erythrocytes are subjected to electrical breakdown in low-ionic-strength media.
Department of Biochemistry and Chemistry, Royal Free Hospital, School of Medicine, University of London, UK.
L Y
LY
Song
Q F
QF
Ahkong
J M
JM
Baldwin
R
R
O'Reilly
J A
JA
Lucy
D002104:Cadmium / Q000737:chemistry; D002413:Cations, Divalent / Q000737:chemistry*; D002459:Cell Fusion / Q000187:drug effects; D048429:Cell Size; D003035:Cobalt / Q000737:chemistry; D004558:Electric Stimulation; D004912:Erythrocytes / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D015726:Giant Cells; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D009994:Osmolar Concentration; D010718:Phosphatidylserines / Q000032:analysis; D010743:Phospholipids / Q000737:chemistry*; D013925:Thromboplastin; D015032:Zinc / Q000737:chemistry* / Q000378:metabolism
D002413:Cations, Divalent; D010718:Phosphatidylserines; D010743:Phospholipids; D002104:Cadmium; D003035:Cobalt; D013925:Thromboplastin; D015032:Zinc