Secretory organelles of Paramecium cells (trichocysts) are not remarkably acidic compartments.
Acridine orange (AO) trapping in conjunction with fluorescence microscopy was applied to Paramecium cells. Trichocysts were not labeled when analyzed with an image intensification system (as opposed to a lysosomal population). Only with increasing intensity of ultraviolet light (UV) did trichocysts (and to some extent the cytosol) exhibit orange fluorescence, both effects being paralleled by increasing cell damage. Therefore, in comparison with the reported cytosolic pH (6.8), trichocysts cannot be considered as essentially acidic compartments. This is supported by experiments in vitro, using isolated cortex fragments or isolated fractions of membrane-bounded trichocysts (greater than or equal to 90% non-leaky). Again, during UV illumination orange fluorescence was observed even in the absence of ATP and Mg2+. Furthermore, this AO fluorescence and the condensation state of trichocyst contents were not affected by NH3 or by any of the widely differing ion- and H(+)-exchange inhibitors or ionophores tested. Decondensation of trichocyst contents occurred only when Ca2+ ionophore A23187 or X537A was incorporated into trichocyst membranes and when Ca2+ was then added. In this case all trichocysts partially decondensed within their intact membranes. We conclude that AO might be trapped in trichocysts by the abundant acidic secretory components during observation with UV light, rather than by acidic luminal pH.
Faculty of Biology, University of Konstanz, Federal Republic of Germany.
C J
CJ
Lumpert
R
R
Glas-Albrecht
E
E
Eisenmann
H
H
Plattner
D000165:Acridine Orange / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D006863:Hydrogen-Ion Concentration; D007091:Image Processing, Computer-Assisted; D007473:Ion Channels / Q000378:metabolism; D008856:Microscopy, Fluorescence; D015388:Organelles / Q000378:metabolism* / Q000648:ultrastructure; D010247:Paramecium / Q000166:cytology / Q000378:metabolism*; D014466:Ultraviolet Rays / Q000009:adverse effects
D007473:Ion Channels; D000165:Acridine Orange