Wp. Schilling et al., Maitotoxin and P2Z/P2X(7) purinergic receptor stimulation activate a common cytolytic pore, AM J P-CELL, 277(4), 1999, pp. C766-C776
The effects of maitotoxin (MTX) on plasmalemma permeability are similar to
these caused by stimulation of P2Z/P2X(7) ionotropic receptors, suggesting
that 1) MTX directly activates P2Z/P2X(7) receptors or 2) MTX and P2Z/P2X(7
) receptor stimulation activate a common cytolytic pore. To distinguish bet
ween these two possibilities, the effect of MTX was examined in 1) THP-1 mo
nocytic cells before and after treatment with lipopolysaccharide and interf
eron-gamma, a maneuver known to upregulate P2Z/P2X(7) receptor, 2) wild-typ
e HEK cells and HEK cells stably expressing the P2Z/P2X(7) receptor, and 3)
BW5147.3 lymphoma cells, a cell line that expresses functional P2Z/P2X(7)
channels that are poorly linked to pore formation. In control THP-1 monocyt
es, addition of MTX produced a biphasic increase in the cytosolic free Ca2 concentration ([Ca2+](i)); the initial increase reflects MTX-induced Ca2influx, whereas the second phase correlates in time with the appearance of
large pores and the uptake of ethidium. MTX produced comparable increases i
n [Ca2+](i) and ethidium uptake in THP-1 monocytes overexpressing the P2Z/P
2X(7) receptor. In both wild-type HEK and HEK cells stably expressing the P
2Z/P2X(7) receptor, MTX-induced increases in [Ca2+](i) and ethidium uptake
were virtually identical. The response of BW5147.3 cells to concentrations
of MTX that produced large increases in [Ca2+](i) had no effect on ethidium
uptake. In both THP-1 and HEK cells, MTX- and Bz-ATP-induced pores activat
e with similar kinetics and exhibit similar size exclusion. Last, MTX-induc
ed pore formation, but not channel activation, is greatly attenuated by red
ucing the temperature to 22 degrees C, a characteristic shared by the P2Z/P
2X(7)-induced pore. Together, the results demonstrate that, although MTX ac
tivates channels that are distinct from those activated by P2Z/P2X(7) recep
tor stimulation, the cytolytic/oncotic pores activated by MTX- and Bz-ATP a
re indistinguishable.