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Cell-specific association of heat shock-induced proton flux with actin ring formation in Chenopodium cells: comparison of auto- and heterotroph cultures

Chaidee, Anchalee, Foissner, Ilse, Pfeiffer, Wolfgang
Protoplasma 2008 v.234 no.1-4 pp. 33-50
Chenopodium rubrum, actin, alkalinization, buffering capacity, calcium, chelating agents, cytochalasin D, ethylene glycol tetraacetic acid, heat, heterotrophs, microfilaments, nigericin, pH, sodium, temperature
A comparison of the responses of extracellular pH, buffering capacity and actin cytoskeleton in autotroph and heterotroph Chenopodium rubrum cells to heat shock revealed cell-specific reactions: alkalinization caused by the heat shock at 25-35°C was higher in heterotroph cells and characterized by heat shock-induced changes in the actin cytoskeleton and ring formation at 35-37°C. Rings (diameter up to 3 μm) disappeared and extracellular pH recovered after the heat-shocked cells were transferred into control medium. At 41°C, no rings but a network of coarse actin filaments were induced; at higher temperatures, fragmentation of the actin cytoskeleton and release of buffering compounds occurred, indicating sudden membrane leakage at 45-47°C. The calcium chelator EGTA [ethylene-glycol-bis(β-aminoethyl-ether)-N,N,N',N'-tetraacetic-acid] increased the frequency of heat shock-induced rings. Ionophore (10 μM nigericin) and the sodium/proton antiport blocker [100 μM 5-(N-ethyl-N-isopropyl)-amiloride] mimicked the effect of the 37°C heat shock. The cytoskeleton inhibitors latrunculin B, cytochalasin D and 2,3-butanedione monoxime inhibited ring formation but not alkalinization. In autotroph cells, the treatment with nigericin (10 μM) produced rings, although the actin cytoskeleton was not affected by temperatures up to 45°C. We conclude that Chenopodium cells express a specific temperature sensor that has ascendancy over the organization of the actin cytoskeleton; this is probably a temperature- and potential-sensitive proton-transporting mechanism that is dependent on the culture conditions of the heterotroph cells.