title: Contribution of K(ir)2 potassium channels to ATP-induced cell death in brain capillary endothelial cells and reconstructed HEK293 cell model.
abstract: Cellular turnover of brain capillary endothelial cells (BCECs) by the balance of cell proliferation and death is essential for maintaining the homeostasis of the blood-brain barrier. Stimulation of metabotropic ATP receptors (P2Y) transiently increased intracellular Ca²(+) concentration ([Ca²(+)](i)) in t-BBEC 117, a cell line derived from bovine BCECs. The [Ca²(+)](i) rise induced membrane hyperpolarization via the activation of apamin-sensitive small-conductance Ca²(+)-activated K(+) channels (SK2) and enhanced cell proliferation in t-BBEC 117. Here, we found anomalous membrane hyperpolarization lasting for over 10 min in response to ATP in ∼15% of t-BBEC 117, in which inward rectifier K(+) channel (K(ir)2.1) was extensively expressed. Once anomalous hyperpolarization was triggered by ATP, it was removed by Ba²(+) but not by apamin. Prolonged exposure to ATPγS increased the relative population of t-BBEC 117, in which the expression of K(ir)2.1 mRNAs was significantly higher and Ba²(+)-sensitive anomalous hyperpolarization was observed. The cultivation of t-BBEC 117 in serum-free medium also increased this population and reduced the cell number. The reduction of cell number was enhanced by the addition of ATPγS and the enhancement was antagonized by Ba²(+). In the human embryonic kidney 293 cell model, where SK2 and K(ir)2.1 were heterologously coexpressed, [Ca²(+)](i) rise by P2Y stimulation triggered anomalous hyperpolarization and cell death. In conclusion, P2Y stimulation in BCECs enhances cell proliferation by SK2 activation in the majority of cells but also triggers cell death in a certain population showing a substantial expression of K(ir)2.1. This dual action of P2Y stimulation may effectively facilitate BCEC turnover.
authors.affiliation: Department of Molecular and Cellular Pharmacology, Nagoya City University, Japan.
authors.forename: Daiju
authors.initials: D
authors.lastname: Yamazaki
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kito
authors.forename: Seiji
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Susumu
authors.initials: S
authors.lastname: Ohya
authors.forename: Hisao
authors.initials: H
authors.lastname: Yamamura
authors.forename: Kiyofumi
authors.initials: K
authors.lastname: Asai
authors.forename: Yuji
authors.initials: Y
authors.lastname: Imaizumi
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001921:Brain / Q000098:blood supply; D002118:Calcium / Q000378:metabolism; D002196:Capillaries; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D008564:Membrane Potentials / Q000502:physiology; D024661:Potassium Channels, Inwardly Rectifying / Q000235:genetics / Q000378:metabolism*; D051640:Small-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000378:metabolism
chemical_list: C496897:KCNN2 protein, human; D024661:Potassium Channels, Inwardly Rectifying; D051640:Small-Conductance Calcium-Activated Potassium Channels; D000255:Adenosine Triphosphate; D002118:Calcium