Ion transport regulated by protease-activated receptor 2 in human airway Calu-3 epithelia

Shinji Sato, Yasushi Ito, Masashi Kondo, Takamasa Ohashi, Satoru Ito, Shinsuke Nakayama, Kaoru Shimokata, Hiroaki Kume

Research output: Contribution to journalArticle

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Abstract

We examined the mechanisms underlying anion secretion mediated by protease-activated receptor 2 (PAR2) and its role in the regulation of ion transport, using polarized human airway Calu-3 cells. PAR2 stimulation by trypsin and a PAR2-activating peptide (PAR2AP), especially from the basolateral aspect, caused transient Cl - secretion due to cytosolic Ca 2+ mobilization. Antagonists of PI-PLC (U73122, ET-18-OCH 3 ) and inositol 1,4,5-triphosphate (xestospongin C (Xest C)) were without effect on the PAR2AP-mediated Cl - secretion, whereas it was attenuated by D609 (a PC-PLC inhibitor) and phorbol 12-myristate 13 acetate (PMA, a PKC activator). Even 30 min after removal of PAR2AP after a 10-min-exposure, cells were still poorly responsive to PAR2 stimulation, but the reduced responsiveness was upregulated by a PKC inhibitor, GF109203X (GFX). Pretreatment with PAR2AP did not affect responses to anion secretagogues, such as isoproterenol, forskolin, thapsigargin, 1-ethyl-2-benzimdazolinone, and adenosine, but ATP-induced responses were significantly reduced. Nystatin permeabilization studies revealed that the presence of PAR2AP prevented ATP-induced increments in basolateral membrane K + conductance without affecting apical membrane Cl - conductance. ATP-elicited Ca 2+ mobilization, which was sensitive to D609 and PMA, was inhibited by the pretreatment with PAR2AP, and this inhibition was blunted by the presence of GFX. Collectively, stimulation of PAR2 generates a brief response of Cl - secretion through PC-PLC-mediated pathway, followed by not only auto-desensitization of PAR2 itself but also cross-desensitization of a PC-PLC-coupled purinoceptor. The two types of desensitization seem likely to have PKC-mediated downregulation of PC-PLC in common.

Original languageEnglish
Pages (from-to)397-407
Number of pages11
JournalBritish Journal of Pharmacology
Volume146
Issue number3
DOIs
Publication statusPublished - 01-10-2005

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PAR-2 Receptor
Ion Transport
Epithelium
Adenosine Triphosphate
Anions
Purinergic Receptors
Nystatin
Inositol 1,4,5-Trisphosphate
Thapsigargin
Membranes
Colforsin
Isoproterenol
Adenosine
Trypsin
Acetates
Down-Regulation
2-furoyl-LIGRLO-amide
Peptides
phosphatidylcholine-specific phospholipase C

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

Sato, Shinji ; Ito, Yasushi ; Kondo, Masashi ; Ohashi, Takamasa ; Ito, Satoru ; Nakayama, Shinsuke ; Shimokata, Kaoru ; Kume, Hiroaki. / Ion transport regulated by protease-activated receptor 2 in human airway Calu-3 epithelia. In: British Journal of Pharmacology. 2005 ; Vol. 146, No. 3. pp. 397-407.
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abstract = "We examined the mechanisms underlying anion secretion mediated by protease-activated receptor 2 (PAR2) and its role in the regulation of ion transport, using polarized human airway Calu-3 cells. PAR2 stimulation by trypsin and a PAR2-activating peptide (PAR2AP), especially from the basolateral aspect, caused transient Cl - secretion due to cytosolic Ca 2+ mobilization. Antagonists of PI-PLC (U73122, ET-18-OCH 3 ) and inositol 1,4,5-triphosphate (xestospongin C (Xest C)) were without effect on the PAR2AP-mediated Cl - secretion, whereas it was attenuated by D609 (a PC-PLC inhibitor) and phorbol 12-myristate 13 acetate (PMA, a PKC activator). Even 30 min after removal of PAR2AP after a 10-min-exposure, cells were still poorly responsive to PAR2 stimulation, but the reduced responsiveness was upregulated by a PKC inhibitor, GF109203X (GFX). Pretreatment with PAR2AP did not affect responses to anion secretagogues, such as isoproterenol, forskolin, thapsigargin, 1-ethyl-2-benzimdazolinone, and adenosine, but ATP-induced responses were significantly reduced. Nystatin permeabilization studies revealed that the presence of PAR2AP prevented ATP-induced increments in basolateral membrane K + conductance without affecting apical membrane Cl - conductance. ATP-elicited Ca 2+ mobilization, which was sensitive to D609 and PMA, was inhibited by the pretreatment with PAR2AP, and this inhibition was blunted by the presence of GFX. Collectively, stimulation of PAR2 generates a brief response of Cl - secretion through PC-PLC-mediated pathway, followed by not only auto-desensitization of PAR2 itself but also cross-desensitization of a PC-PLC-coupled purinoceptor. The two types of desensitization seem likely to have PKC-mediated downregulation of PC-PLC in common.",
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Sato, S, Ito, Y, Kondo, M, Ohashi, T, Ito, S, Nakayama, S, Shimokata, K & Kume, H 2005, 'Ion transport regulated by protease-activated receptor 2 in human airway Calu-3 epithelia', British Journal of Pharmacology, vol. 146, no. 3, pp. 397-407. https://doi.org/10.1038/sj.bjp.0706330

Ion transport regulated by protease-activated receptor 2 in human airway Calu-3 epithelia. / Sato, Shinji; Ito, Yasushi; Kondo, Masashi; Ohashi, Takamasa; Ito, Satoru; Nakayama, Shinsuke; Shimokata, Kaoru; Kume, Hiroaki.

In: British Journal of Pharmacology, Vol. 146, No. 3, 01.10.2005, p. 397-407.

Research output: Contribution to journalArticle

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T1 - Ion transport regulated by protease-activated receptor 2 in human airway Calu-3 epithelia

AU - Sato, Shinji

AU - Ito, Yasushi

AU - Kondo, Masashi

AU - Ohashi, Takamasa

AU - Ito, Satoru

AU - Nakayama, Shinsuke

AU - Shimokata, Kaoru

AU - Kume, Hiroaki

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N2 - We examined the mechanisms underlying anion secretion mediated by protease-activated receptor 2 (PAR2) and its role in the regulation of ion transport, using polarized human airway Calu-3 cells. PAR2 stimulation by trypsin and a PAR2-activating peptide (PAR2AP), especially from the basolateral aspect, caused transient Cl - secretion due to cytosolic Ca 2+ mobilization. Antagonists of PI-PLC (U73122, ET-18-OCH 3 ) and inositol 1,4,5-triphosphate (xestospongin C (Xest C)) were without effect on the PAR2AP-mediated Cl - secretion, whereas it was attenuated by D609 (a PC-PLC inhibitor) and phorbol 12-myristate 13 acetate (PMA, a PKC activator). Even 30 min after removal of PAR2AP after a 10-min-exposure, cells were still poorly responsive to PAR2 stimulation, but the reduced responsiveness was upregulated by a PKC inhibitor, GF109203X (GFX). Pretreatment with PAR2AP did not affect responses to anion secretagogues, such as isoproterenol, forskolin, thapsigargin, 1-ethyl-2-benzimdazolinone, and adenosine, but ATP-induced responses were significantly reduced. Nystatin permeabilization studies revealed that the presence of PAR2AP prevented ATP-induced increments in basolateral membrane K + conductance without affecting apical membrane Cl - conductance. ATP-elicited Ca 2+ mobilization, which was sensitive to D609 and PMA, was inhibited by the pretreatment with PAR2AP, and this inhibition was blunted by the presence of GFX. Collectively, stimulation of PAR2 generates a brief response of Cl - secretion through PC-PLC-mediated pathway, followed by not only auto-desensitization of PAR2 itself but also cross-desensitization of a PC-PLC-coupled purinoceptor. The two types of desensitization seem likely to have PKC-mediated downregulation of PC-PLC in common.

AB - We examined the mechanisms underlying anion secretion mediated by protease-activated receptor 2 (PAR2) and its role in the regulation of ion transport, using polarized human airway Calu-3 cells. PAR2 stimulation by trypsin and a PAR2-activating peptide (PAR2AP), especially from the basolateral aspect, caused transient Cl - secretion due to cytosolic Ca 2+ mobilization. Antagonists of PI-PLC (U73122, ET-18-OCH 3 ) and inositol 1,4,5-triphosphate (xestospongin C (Xest C)) were without effect on the PAR2AP-mediated Cl - secretion, whereas it was attenuated by D609 (a PC-PLC inhibitor) and phorbol 12-myristate 13 acetate (PMA, a PKC activator). Even 30 min after removal of PAR2AP after a 10-min-exposure, cells were still poorly responsive to PAR2 stimulation, but the reduced responsiveness was upregulated by a PKC inhibitor, GF109203X (GFX). Pretreatment with PAR2AP did not affect responses to anion secretagogues, such as isoproterenol, forskolin, thapsigargin, 1-ethyl-2-benzimdazolinone, and adenosine, but ATP-induced responses were significantly reduced. Nystatin permeabilization studies revealed that the presence of PAR2AP prevented ATP-induced increments in basolateral membrane K + conductance without affecting apical membrane Cl - conductance. ATP-elicited Ca 2+ mobilization, which was sensitive to D609 and PMA, was inhibited by the pretreatment with PAR2AP, and this inhibition was blunted by the presence of GFX. Collectively, stimulation of PAR2 generates a brief response of Cl - secretion through PC-PLC-mediated pathway, followed by not only auto-desensitization of PAR2 itself but also cross-desensitization of a PC-PLC-coupled purinoceptor. The two types of desensitization seem likely to have PKC-mediated downregulation of PC-PLC in common.

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