Synchronous exocytosis in Paramecium cells. IV. Polyamino compounds as potent trigger agents for repeatable...

We found that different polyamino compounds trigger the instantaneous and simultaneous release of trichocysts from Paramecium tetraurelia cells (monoxenically bacterized cultures), provided amino groups are spaced in intervals of approximately 1.0 nm; in this case even diamines or oligopeptides exert some trigger effect. The most potent trigger agent found was aminoethyldextran (AED, MW = 40 kDa) which was used mainly as a derivative with 40 -NH2/molecule. A maximal response (strain K401) was achieved at 1.38 X 10(-6) M, a half maximal response at 1.08 X 10(-6) M. AED acts by a dual effect, i.e., not only by statistically properly spaced amino groups but also by some additional effect of the dextran molecule, since this greatly enhances the effect of oligoamines, although it exerts no trigger effect per se. From a total of approximately 1120 or approximately 1230 trichocysts per cell (strain K401 or 7S) approximately 95% are releasable by AED. In these strains the number of non-releasable trichocysts corresponds closely to the number of undocked trichocysts floating in the cytoplasm, so that practically all trichocysts which are docked to the cell membrane, can be released. (We also analysed different mutant strains for their response to AED.) Massive trichocyst release does not impair cell viability or culture growth, and multiple release-redocking cycles can be performed; up to 5 trigger-docking cycles were tested with individual cells in 12 h intervals. AED-triggered exocytosis requires a free extracellular [Ca2+] of greater than or equal to 10(-5) M; it is inhibited by EGTA (ethyleneglycol-bis(beta-aminoethyl ether)-N,N'-tetraacetate), by a short pH 5.5 shock or by neomycin at 10(-5) M concentration.(ABSTRACT TRUNCATED AT 250 WORDS)