Bradykinin-evoked Ca2+ mobilization in Madin Darby canine kidney cells

Chung Ren Jan, Chin Man Ho, Sheng Nan Wu, Ching Jiunn Tseng

Research output: Contribution to journalJournal Article peer-review

81 Scopus citations

Abstract

We studied the mechanisms underlying the bradykinin-evoked changes in intracellular calcium concentration ([Ca2+](i)) in Madin Darby canine kidney (MDCK) cells. Bradykinin evoked a [Ca2+](i) transient in a dose-dependent manner, measured by fura-2 fluorimetry and digital video imaging. The transient consisted of a rise and a decay and [Ca2+](i) returned to baseline without oscillations. External Ca2+ influx occurred, as demonstrated by Mn2+ quench and external Ca2+ removal measurements. Bradykinin acted by stimulating bradykinin B2 receptors as evidenced by blockade by d-arginyl-l-arginlyl-l-prolyl-trans-4-hydroxy-l-prolylglycyl-3-(2-thienyl)-l-alanyl-l-seryl-d-1,2,3,4-tetrahydro-3-isoquinolinecarbonyl-l-(2α,3β,7aβ)-octahydro-1H-indole-2-carbonyl-l-arginine (HOE 140) but not by d-arginyl-l-arginlyl-l-prolyl-trans-4-hydroxy-l-prolylglycyl-3-(2-thienyl)-l-alanyl-l-seryl-d-1,2,3,4-tetrahydro-3-isoquinolinecarbonyl-l-(2α,3β,7aβ)-octahydro-1H-indole-2-carbonyl ([Des-Arg]HOE 140). The [Ca2+](i) signal was abolished by 1-(6-((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122) and partially inhibited by neomycin, implying mediation by phospholipase C. The transient was initiated by a release of Ca2+ from internal stores since it was abolished by pretreatment with thapsigargin or cyclopiazonic acid. The mobilization of the internal Ca2+ store subsequently triggered a 1-[β-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole hydrochloride (SKF 96365)-insensitive Ca2+ entry. Pretreatment with carbonylcyanide m-chlorophynylhydrozone and gly-phe-β-naphthylamide did not alter the transient, thus excluding the participation of mitochondria and lysosomes. Efflux via Ca2+ pumps contributed to the decay of the transient. Efflux via Na+/Ca2+ exchange or sequestration by mitochondria and lysosomes was insignificant. The transient was blunted by the protein kinase C activator phorbol 12-myristate 13-acetate, and was enhanced by the protein kinase C inhibitors sphingosine and chelerythrine, the protein kinase A inhibitor 2,5-di-(t-butyl)-1,4-hydroquinone, N-[2-(p-bromocinnamylamino)ethyl]5-isoquinolinesulfonamide (H-89), the agent 8-(diethylamino)octyl 3,4,5-trimethoxybenzoate (TMB-8), and agents that elevated levels of 3',5'-cyclic guanosine monophosphate. The transient did not heterologously desensitize with that evoked by ATP, ADP or UTP. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)219-233
Number of pages15
JournalEuropean Journal of Pharmacology
Volume355
Issue number2-3
DOIs
StatePublished - 21 08 1998
Externally publishedYes

Keywords

  • Bradykinin
  • Ca stores
  • Ca, thapsigargin
  • Fura-2
  • MDCK cells

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