An exocytotic mutant of Paramecium caudatum: membrane fusion without secretory contents release

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1998
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Klauke, Norbert
Haga, Nobuyuki
Watanabe, Tsuyoshi
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Cell Calcium. 1998, 23(5), pp. 349-360. ISSN 0143-4160. Available under: doi: 10.1016/S0143-4160(98)90030-6
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This is a detailed characterization of a secretory mutant incapable of releasing secretory contents despite normal exocytotic membrane fusion performance. Trichocyst non-discharge strain tnd1 of Paramecium caudatum and its wildtype (wt) both show a transient cortical [Ca2+]i increase and exocytotic membrane fusion in response to the polyamine secretagogue, aminoethyldextran (AED), or to caffeine. tnd1 cells frequently display spontaneous Ca2+ signals parallelled by spontaneous exocytotic membrane fusion. This remains undetected, unless the trichocyst matrix is shown to be freely accessible to the inert, non-membrane permeable fluorochrome, F2FITC, from the outside. In these tnd1 cells, spontaneous and AED- or caffeine-induced membrane fusion, always without contents expulsion by decondensation (i.e. several-fold stretching), is ascertained by electron microscopy. Exocytotic openings, with condensed trichocysts retained, may persist for hours without impairing cells. Trichocyst decondensation normally requires micromolar [Ca2+]e, but an increase to 10 mM has no effect on tnd1 trichocyst expansion in vivo or in vitro (when isolated and exposed to ionophore A23187 + Ca2+). Paracrystalline packing of the major secretory components (trichynins) does occur, despite incomplete proteolytic precursor processing (according to SDS-PAGE). However, 45Ca(2+)-binding by secretory components is considerably reduced--the likely cause of the non-discharge phenotype. Our findings imply significant untriggered membrane fusion in a system normally following the triggered pathway and clear separation of exocytotic membrane fusion from any later Ca(2+)-dependent steps of the secretory cycle.

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ISO 690KLAUKE, Norbert, Roland KISSMEHL, Helmut PLATTNER, Nobuyuki HAGA, Tsuyoshi WATANABE, 1998. An exocytotic mutant of Paramecium caudatum: membrane fusion without secretory contents release. In: Cell Calcium. 1998, 23(5), pp. 349-360. ISSN 0143-4160. Available under: doi: 10.1016/S0143-4160(98)90030-6
BibTex
@article{Klauke1998exocy-6830,
  year={1998},
  doi={10.1016/S0143-4160(98)90030-6},
  title={An exocytotic mutant of Paramecium caudatum: membrane fusion without secretory contents release},
  number={5},
  volume={23},
  issn={0143-4160},
  journal={Cell Calcium},
  pages={349--360},
  author={Klauke, Norbert and Kissmehl, Roland and Plattner, Helmut and Haga, Nobuyuki and Watanabe, Tsuyoshi}
}
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