Skip to main content
placeholder image

Calcium entry-dependent oscillations of cytoplasmic calcium concentration in cultured endothelial cell monolayers

Journal Article


Abstract


  • Bovine endothelial cell monolayers grown to confluence and stimulated with bradykinin responded with periodic fluctuations in intracellular Ca2+ concentration ([Ca2+]i) when exposed to K+-free Hepes-buffered saline. The fluctuations in [Ca2+]i measured with fura-2 were synchronized among the population of cells observed and were sensitive to extracellular Ca2+ concentration ([Ca2+]o). Thapsigargin, which inhibits the endoplasmic reticular Ca2+-ATPase, did not inhibit the [Ca2+]i oscillations. Removal of extracellular Ca2+ or inhibition of Ca2+ entry by using La3+ or l-{β-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl}-1H-imidazole hydrochloride (SKF 96365) abolished the [Ca2+]i oscillations in endothelial cell monolayers. The fluctuations in [Ca2+]i were therefore dependent on Ca2+ influx rather than Ca2+ mobilization from intracellular stores. Simultaneous measurements of membrane potential (Em) using the potential-sensitive bisoxonol dye bis(1,3-dibutylbarbituric acidltrimethine oxonol [Di-BAC4(3)] and [Ca2+]i using fura-2 showed that Em oscillated at the same frequency as the fluctuations in [Ca2+]i. The peak depolarization signal coincided with the maximum rate of increase in the [Ca2+]i signal. Oscillations in the Em signal were inhibited by removal of Ca2+ or by addition of 1 mM Ni2+ to the external solution. Taken together, these observations suggest that the change in Em is the consequence of oscillatory changes in a membrane conductance that also allows Ca2+ to enter the cell. Oscillations in the DiBAC4(3) signal may reflect a rhythmic entry of Ca2+ through nonselective cation channels.

UOW Authors


  •   Laskey, Rachel E. (external author)
  •   Adams, David
  •   Cannell, M (external author)
  •   Van Breemen, Cornelis (external author)

Publication Date


  • 1992

Citation


  • Laskey, R. E., Adams, D. J., Cannell, M. & Van Breemen, C. (1992). Calcium entry-dependent oscillations of cytoplasmic calcium concentration in cultured endothelial cell monolayers. Proceedings of the National Academy of Sciences of USA, 89 (5), 1690-1694.

Scopus Eid


  • 2-s2.0-0026531136

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/735

Number Of Pages


  • 4

Start Page


  • 1690

End Page


  • 1694

Volume


  • 89

Issue


  • 5

Place Of Publication


  • http://www.ncbi.nlm.nih.gov/pmc/articles/PMC48518/

Abstract


  • Bovine endothelial cell monolayers grown to confluence and stimulated with bradykinin responded with periodic fluctuations in intracellular Ca2+ concentration ([Ca2+]i) when exposed to K+-free Hepes-buffered saline. The fluctuations in [Ca2+]i measured with fura-2 were synchronized among the population of cells observed and were sensitive to extracellular Ca2+ concentration ([Ca2+]o). Thapsigargin, which inhibits the endoplasmic reticular Ca2+-ATPase, did not inhibit the [Ca2+]i oscillations. Removal of extracellular Ca2+ or inhibition of Ca2+ entry by using La3+ or l-{β-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl}-1H-imidazole hydrochloride (SKF 96365) abolished the [Ca2+]i oscillations in endothelial cell monolayers. The fluctuations in [Ca2+]i were therefore dependent on Ca2+ influx rather than Ca2+ mobilization from intracellular stores. Simultaneous measurements of membrane potential (Em) using the potential-sensitive bisoxonol dye bis(1,3-dibutylbarbituric acidltrimethine oxonol [Di-BAC4(3)] and [Ca2+]i using fura-2 showed that Em oscillated at the same frequency as the fluctuations in [Ca2+]i. The peak depolarization signal coincided with the maximum rate of increase in the [Ca2+]i signal. Oscillations in the Em signal were inhibited by removal of Ca2+ or by addition of 1 mM Ni2+ to the external solution. Taken together, these observations suggest that the change in Em is the consequence of oscillatory changes in a membrane conductance that also allows Ca2+ to enter the cell. Oscillations in the DiBAC4(3) signal may reflect a rhythmic entry of Ca2+ through nonselective cation channels.

UOW Authors


  •   Laskey, Rachel E. (external author)
  •   Adams, David
  •   Cannell, M (external author)
  •   Van Breemen, Cornelis (external author)

Publication Date


  • 1992

Citation


  • Laskey, R. E., Adams, D. J., Cannell, M. & Van Breemen, C. (1992). Calcium entry-dependent oscillations of cytoplasmic calcium concentration in cultured endothelial cell monolayers. Proceedings of the National Academy of Sciences of USA, 89 (5), 1690-1694.

Scopus Eid


  • 2-s2.0-0026531136

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/735

Number Of Pages


  • 4

Start Page


  • 1690

End Page


  • 1694

Volume


  • 89

Issue


  • 5

Place Of Publication


  • http://www.ncbi.nlm.nih.gov/pmc/articles/PMC48518/