Activation of the cGMP/PKG pathway inhibits electrical activity in rabbit urethral interstitial cells of Cajal by reducing the spatial spread of Ca 2+ waves

Sergeant, Gerard P. and Johnston, Louise and McHale, Noel G. and Hollywood, Mark A. (2006) Activation of the cGMP/PKG pathway inhibits electrical activity in rabbit urethral interstitial cells of Cajal by reducing the spatial spread of Ca 2+ waves. Journal of Physiology, 574 (1). pp. 167-181.

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Abstract

In the present study we used a combination of patch clamping and fast confocal Ca 2+ imaging to examine the effects of activators of the nitric oxide (NO)/cGMP pathway on pacemaker activity in freshly dispersed ICC from the rabbit urethra, using the amphotericin B perforated patch configuration of the patch-clamp technique. The nitric oxide donor, DEA-NO, the soluble guanylyl cyclase activator YC-1 and the membrane-permeant analogue of cGMP, 8-Br-cGMP inhibited spontaneous transient depolarizations (STDs) and spontaneous transient inward currents (STICs) recorded under current-clamp and voltageclamp conditions, respectively. Caffeine-evoked Cl – currents were unaltered in the presence of SP-8-Br-PET-cGMPs, suggesting that activation of the cGMP/PKG pathway does not block Cl – channels directly or interfere with Ca 2+ release via ryanodine receptors (RyR). However, noradrenaline-evoked Cl – currents were attenuated by SP-8-Br-PET-cGMPs, suggesting that activation of cGMP-dependent protein kinase (PKG) may modulate release of Ca 2+ via IP 3 receptors (IP 3 R).When urethral interstitial cells (ICC) were loaded with Fluo4-AM (2 μM), and viewed with a confocal microscope, they fired regular propagating Ca 2+ waves, which originated in one or more regions of the cell. Application of DEA-NO or other activators of the cGMP/PKG pathway did not significantly affect the oscillation frequency of these cells, but did significantly reduce their spatial spread. These effects were mimicked by the IP 3 R blocker, 2-APB (100 μM). These data suggest that NO donors and activators of the cGMP pathway inhibit electrical activity of urethral ICC by reducing the spatial spread of Ca 2+ waves, rather than decreasing wave frequency.

Item Type:	Article
Uncontrolled Keywords:	Muscle, Smooth; Patch clamp techniques; Interstitial cells of Cajal; Urethra; Rabbit
Subjects:	Science
Research Centres:	Smooth Muscle Research Centre
Depositing User:	Concepta Woods
Date Deposited:	26 Nov 2011 12:58
Last Modified:	11 Nov 2014 16:10
URI:	https://eprints.dkit.ie/id/eprint/68

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