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Refilling of cortical calcium stores in Paramecium cells : in situ analysis in correlation with store-operated calcium influx

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2003

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Mohamed, Ihab K.
Husser, Marc R.
Sehring, Ivonne Margarete
Hentschel, C.

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Cell Calcium. 2003, 34(1), pp. 87-96. ISSN 0143-4160. Available under: doi: 10.1016/S0143-4160(03)00025-3

Zusammenfassung

This is the first thorough study of refilling of a cortical calcium store in a secretory cell after stimulation in which we combined widely different methodologies. Stimulation of dense-core vesicle ( trichocysts ) exocytosis in Paramecium involves a Ca2+-influx superimposed to Ca2+-release from cortical stores ( alveolar sacs (ASs)). In quenched-flow experiments, membrane fusion frequency rose with increasing [Ca2+]o in the medium, from 20 25% at [Ca2+]o ≤0.25 μM to 100% at [Ca2+]o between 2 and 10 μM, i.e. close to the range of estimated local intracellular [Ca2+] during membrane fusion. Next, we analyzed Ca2+-specific fluorochrome signals during stimulation under different conditions. Treatment with actin-reactive drugs had no effect on Ca2+-signaling. In double trigger experiments, with BAPTA in the second secretagogue application (BAPTA only for stimulation and analysis), the cortical Ca2+-signal (due solely to Ca2+ released from cortical stores) recovered with t1/2 65 min. When ASs were analyzed in situ by X-ray microanalysis after different trigger times (+Ca2+o), t1/2 for store refilling was similar, 60 min. These values are similar to previously measured 45Ca2+-uptake by isolated ASs. In sum we find, (i) exogenous Ca2+ increases exocytosis/membrane fusion performance with EC50=0.7 μM, (ii) Ca2+-signaling in this system is not sensitive to actin-reactive drugs, and (iii) refilling of these cortical calcium stores goes on over hours and thus is much slower than expected.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie

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Calcium, Ciliates, Exocytosis, Paramecium, Secretion

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ISO 690MOHAMED, Ihab K., Marc R. HUSSER, Ivonne Margarete SEHRING, Joachim HENTSCHEL, C. HENTSCHEL, Helmut PLATTNER, 2003. Refilling of cortical calcium stores in Paramecium cells : in situ analysis in correlation with store-operated calcium influx. In: Cell Calcium. 2003, 34(1), pp. 87-96. ISSN 0143-4160. Available under: doi: 10.1016/S0143-4160(03)00025-3
BibTex
@article{Mohamed2003Refil-8529,
  year={2003},
  doi={10.1016/S0143-4160(03)00025-3},
  title={Refilling of cortical calcium stores in Paramecium cells : in situ analysis in correlation with store-operated calcium influx},
  number={1},
  volume={34},
  issn={0143-4160},
  journal={Cell Calcium},
  pages={87--96},
  author={Mohamed, Ihab K. and Husser, Marc R. and Sehring, Ivonne Margarete and Hentschel, Joachim and Hentschel, C. and Plattner, Helmut}
}
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