Kinetics of Empty Store-activated Ca2+ Influx in HeLa Cells

Ludwig Missiaen, Humbert De Smedt, Jan B. Pary, Masahiro Oike, Rik Casteels

Research output: Contribution to journalArticle

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Abstract

The intracellular Ca2+ indicator Indo-1 was used to monitor changes in cytosolic [Ca2+] ([Ca2+]i) in single HeLa cells upon readmission of external Ca2+ after a short incubation in Ca2+-free solution. HeLa cells were responsive to histamine but not to caffeine, and their histamine-sensitive store was totally depleted by a 60-min exposure to 2 μM thapsigargin. The resting [Ca2+]i in thapsigargin-treated cells was higher than in control cells and low amplitude [Ca2+]i oscillations were observed in about 20% of the cells. Readmission of external Ca2+ after a brief withdrawal of extracellular Ca2+ resulted in a transient [Ca2+]i rise, which then decayed to the same elevated [Ca2+]i measured before the Ca2+ withdrawal period. The [Ca2+]i rise was associated with an increased rate of Mn2+ entry, measured as the rate of quenching of intracellular Fura-2. The same procedure did not affect the [Ca2+]i in control cells not pretreated with thapsigargin. The amplitude of this [Ca2+]i transient in thapsigargin pretreated cells depended on the duration of prior incubation in Ca2+-free medium. The [Ca2+]i rise induced by elevating the extracellular [Ca2+] from 1.5 to 10 mM was more pronounced if the [Ca2+]i during the initial incubation in 1.5 mM Ca2+ was first lowered by depolarizing the cells. We conclude that an empty store stimulates a Ca2+ entry pathway consisting of two components: a continuously elevated basal leak and a second component that is transient due to the high [Ca2+]i-induced inhibition of the Ca2+ entry pathway. This inhibition and the subsequent recovery from it as the [Ca2+]i is brought to resting levels could cause the oscillatory Ca2+ entry that we recorded in a fraction of the thapsigargin-treated cells.

Original languageEnglish
Pages (from-to)5817-5823
Number of pages7
JournalJournal of Biological Chemistry
Volume269
Issue number8
Publication statusPublished - Feb 25 1994
Externally publishedYes

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Thapsigargin
HeLa Cells
Kinetics
Histamine
Cells
Fura-2
Caffeine
Quenching
Recovery

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Missiaen, L., De Smedt, H., Pary, J. B., Oike, M., & Casteels, R. (1994). Kinetics of Empty Store-activated Ca2+ Influx in HeLa Cells. Journal of Biological Chemistry, 269(8), 5817-5823.

Kinetics of Empty Store-activated Ca2+ Influx in HeLa Cells. / Missiaen, Ludwig; De Smedt, Humbert; Pary, Jan B.; Oike, Masahiro; Casteels, Rik.

In: Journal of Biological Chemistry, Vol. 269, No. 8, 25.02.1994, p. 5817-5823.

Research output: Contribution to journalArticle

Missiaen, L, De Smedt, H, Pary, JB, Oike, M & Casteels, R 1994, 'Kinetics of Empty Store-activated Ca2+ Influx in HeLa Cells', Journal of Biological Chemistry, vol. 269, no. 8, pp. 5817-5823.
Missiaen L, De Smedt H, Pary JB, Oike M, Casteels R. Kinetics of Empty Store-activated Ca2+ Influx in HeLa Cells. Journal of Biological Chemistry. 1994 Feb 25;269(8):5817-5823.
Missiaen, Ludwig ; De Smedt, Humbert ; Pary, Jan B. ; Oike, Masahiro ; Casteels, Rik. / Kinetics of Empty Store-activated Ca2+ Influx in HeLa Cells. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 8. pp. 5817-5823.
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