Obstruction enhances rho-kinase pathway and diminishes protein kinase C pathway in carbachol-induced calcium sensitization in contraction of α-toxin permeabilized guinea pig detrusor smooth muscle

Nouval Shahab, Shunichi Kajioka, Fumi Takahashi, Mitsuho Onimaru, Miho Matsuda, Narihito Seki, Seiji Naito

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Abstract

Aims We investigated the relative important role of rho kinase (ROK) and protein kinase C (PKC) pathways in carbachol (CCh)-induced Ca 2+ sensitization in α-toxin permeabilized Guinea pig detrusor smooth muscle (DSM) following bladder outlet obstruction (BOO). Methods Bladder outlet obstruction was created by placement of a silver jeweler's jump rings loosely round the urethro-vesical junction of Guinea pigs. Sham operated Guinea pig underwent a similar protocol without application of the ring and served as control. α-Toxin permeabilized DSM strips from control Guinea pigs and those subjected to 6-8 weeks of BOO were mounted horizontally for isometric force recording in 100μl relaxing solution on perspex block. The effect of ROK inhibitor (Y-27632) and PKC inhibitor (GF-109203X) on CCh-induced Ca 2+ sensitization was studied during sustained contraction. Permeabilized DSM strips were also stimulated by cumulative increase of Ca 2+ concentration compared to that in control in the presence and in the absence of sensitization-induced PKC activator, phorbol 12,13-dibutyrate. Results Ca 2+ sensitization-induced by CCh was greater in BOO compared to controls. This muscarinic agonist-induced Ca 2+ sensitization was inhibited by Y-27632 or GF-109203X. The inhibitory effect of Y-27632 (5μM) was greater while the inhibitory effect of GF-109203X (5μM) was smaller in BOO compared to that in controls. Phorbol 12,13-dibutyrate (1μM) markedly increased Ca 2+ sensitivity in controls but not in BOO. Conclusions Our findings provide the first evidence that BOO enhances the ROK pathway and diminishes the PKC pathway in CCh-induced Ca 2+ sensitization in contraction of permeabilized Guinea pig DSM and suggest that inhibitors of ROK might potentially relieve bladder dysfunction related to BOO.

Original languageEnglish
Pages (from-to)593-599
Number of pages7
JournalNeurourology and Urodynamics
Volume31
Issue number4
DOIs
Publication statusPublished - Apr 1 2012

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Urinary Bladder Neck Obstruction
rho-Associated Kinases
Carbachol
Protein Kinase C
Smooth Muscle
Guinea Pigs
Calcium
Phorbol 12,13-Dibutyrate
Urinary Bladder
Muscarinic Agonists
Protein C Inhibitor
Polymethyl Methacrylate
Protein Kinase Inhibitors
Silver

All Science Journal Classification (ASJC) codes

  • Clinical Neurology
  • Urology

Cite this

@article{e66a2e1869484fd5910825ff8ddb47cf,
title = "Obstruction enhances rho-kinase pathway and diminishes protein kinase C pathway in carbachol-induced calcium sensitization in contraction of α-toxin permeabilized guinea pig detrusor smooth muscle",
abstract = "Aims We investigated the relative important role of rho kinase (ROK) and protein kinase C (PKC) pathways in carbachol (CCh)-induced Ca 2+ sensitization in α-toxin permeabilized Guinea pig detrusor smooth muscle (DSM) following bladder outlet obstruction (BOO). Methods Bladder outlet obstruction was created by placement of a silver jeweler's jump rings loosely round the urethro-vesical junction of Guinea pigs. Sham operated Guinea pig underwent a similar protocol without application of the ring and served as control. α-Toxin permeabilized DSM strips from control Guinea pigs and those subjected to 6-8 weeks of BOO were mounted horizontally for isometric force recording in 100μl relaxing solution on perspex block. The effect of ROK inhibitor (Y-27632) and PKC inhibitor (GF-109203X) on CCh-induced Ca 2+ sensitization was studied during sustained contraction. Permeabilized DSM strips were also stimulated by cumulative increase of Ca 2+ concentration compared to that in control in the presence and in the absence of sensitization-induced PKC activator, phorbol 12,13-dibutyrate. Results Ca 2+ sensitization-induced by CCh was greater in BOO compared to controls. This muscarinic agonist-induced Ca 2+ sensitization was inhibited by Y-27632 or GF-109203X. The inhibitory effect of Y-27632 (5μM) was greater while the inhibitory effect of GF-109203X (5μM) was smaller in BOO compared to that in controls. Phorbol 12,13-dibutyrate (1μM) markedly increased Ca 2+ sensitivity in controls but not in BOO. Conclusions Our findings provide the first evidence that BOO enhances the ROK pathway and diminishes the PKC pathway in CCh-induced Ca 2+ sensitization in contraction of permeabilized Guinea pig DSM and suggest that inhibitors of ROK might potentially relieve bladder dysfunction related to BOO.",
author = "Nouval Shahab and Shunichi Kajioka and Fumi Takahashi and Mitsuho Onimaru and Miho Matsuda and Narihito Seki and Seiji Naito",
year = "2012",
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language = "English",
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pages = "593--599",
journal = "Neurourology and Urodynamics",
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T1 - Obstruction enhances rho-kinase pathway and diminishes protein kinase C pathway in carbachol-induced calcium sensitization in contraction of α-toxin permeabilized guinea pig detrusor smooth muscle

AU - Shahab, Nouval

AU - Kajioka, Shunichi

AU - Takahashi, Fumi

AU - Onimaru, Mitsuho

AU - Matsuda, Miho

AU - Seki, Narihito

AU - Naito, Seiji

PY - 2012/4/1

Y1 - 2012/4/1

N2 - Aims We investigated the relative important role of rho kinase (ROK) and protein kinase C (PKC) pathways in carbachol (CCh)-induced Ca 2+ sensitization in α-toxin permeabilized Guinea pig detrusor smooth muscle (DSM) following bladder outlet obstruction (BOO). Methods Bladder outlet obstruction was created by placement of a silver jeweler's jump rings loosely round the urethro-vesical junction of Guinea pigs. Sham operated Guinea pig underwent a similar protocol without application of the ring and served as control. α-Toxin permeabilized DSM strips from control Guinea pigs and those subjected to 6-8 weeks of BOO were mounted horizontally for isometric force recording in 100μl relaxing solution on perspex block. The effect of ROK inhibitor (Y-27632) and PKC inhibitor (GF-109203X) on CCh-induced Ca 2+ sensitization was studied during sustained contraction. Permeabilized DSM strips were also stimulated by cumulative increase of Ca 2+ concentration compared to that in control in the presence and in the absence of sensitization-induced PKC activator, phorbol 12,13-dibutyrate. Results Ca 2+ sensitization-induced by CCh was greater in BOO compared to controls. This muscarinic agonist-induced Ca 2+ sensitization was inhibited by Y-27632 or GF-109203X. The inhibitory effect of Y-27632 (5μM) was greater while the inhibitory effect of GF-109203X (5μM) was smaller in BOO compared to that in controls. Phorbol 12,13-dibutyrate (1μM) markedly increased Ca 2+ sensitivity in controls but not in BOO. Conclusions Our findings provide the first evidence that BOO enhances the ROK pathway and diminishes the PKC pathway in CCh-induced Ca 2+ sensitization in contraction of permeabilized Guinea pig DSM and suggest that inhibitors of ROK might potentially relieve bladder dysfunction related to BOO.

AB - Aims We investigated the relative important role of rho kinase (ROK) and protein kinase C (PKC) pathways in carbachol (CCh)-induced Ca 2+ sensitization in α-toxin permeabilized Guinea pig detrusor smooth muscle (DSM) following bladder outlet obstruction (BOO). Methods Bladder outlet obstruction was created by placement of a silver jeweler's jump rings loosely round the urethro-vesical junction of Guinea pigs. Sham operated Guinea pig underwent a similar protocol without application of the ring and served as control. α-Toxin permeabilized DSM strips from control Guinea pigs and those subjected to 6-8 weeks of BOO were mounted horizontally for isometric force recording in 100μl relaxing solution on perspex block. The effect of ROK inhibitor (Y-27632) and PKC inhibitor (GF-109203X) on CCh-induced Ca 2+ sensitization was studied during sustained contraction. Permeabilized DSM strips were also stimulated by cumulative increase of Ca 2+ concentration compared to that in control in the presence and in the absence of sensitization-induced PKC activator, phorbol 12,13-dibutyrate. Results Ca 2+ sensitization-induced by CCh was greater in BOO compared to controls. This muscarinic agonist-induced Ca 2+ sensitization was inhibited by Y-27632 or GF-109203X. The inhibitory effect of Y-27632 (5μM) was greater while the inhibitory effect of GF-109203X (5μM) was smaller in BOO compared to that in controls. Phorbol 12,13-dibutyrate (1μM) markedly increased Ca 2+ sensitivity in controls but not in BOO. Conclusions Our findings provide the first evidence that BOO enhances the ROK pathway and diminishes the PKC pathway in CCh-induced Ca 2+ sensitization in contraction of permeabilized Guinea pig DSM and suggest that inhibitors of ROK might potentially relieve bladder dysfunction related to BOO.

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U2 - 10.1002/nau.21193

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M3 - Article

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