Cholinergic-induced Ca2+ signaling in interstitial cells of Cajal from the guinea pig bladder

L. Johnston, C. Carson, A.D. Lyons, R.A. Davidson, K.D. McCloskey

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Acetylcholine released from parasympathetic excitatory nerves activates contraction in detrusor smooth muscle. Immunohistochemical labeling of guinea pig detrusor with anti-c-Kit and anti-VAChT demonstrated a close structural relationship between interstitial cells of Cajal (ICC) and cholinergic nerves. The ability of guinea pig bladder detrusor ICC to respond to the acetylcholine analog, carbachol, was investigated in enzymatically dissociated cells, loaded with the Ca2+ indicator fluo 4AM. ICC fired Ca2+ transients in response to stimulation by carbachol (1/10 M). Their pharmacology was consistent with carbachol-induced contractions in strips of detrusor which were inhibited by 4-DAMP (1 M), an M3 receptor antagonist, but not by the M2 receptor antagonist methoctramine (1 M). The source of Ca2+ underlying the carbachol transients in isolated ICC was investigated using agents to interfere with influx or release from intracellular stores. Nifedipine (1 M) or Ni2 (30-100 M) to block Ca2+ channels or the removal of external Ca2 reduced the amplitude of the carbachol transients. Application of ryanodine (30 M) or tetracaine (100 M) abolished the transients. The phospholipase C inhibitor, U-73122 (2.5 M), significantly reduced the responses. 2-Aminoethoxydiethylborate (30 M) caused a significant reduction and Xestospongin C (1 M) was more effective, almost abolishing the responses. Intact in situ preparations of guinea pig bladder loaded with a Ca2+ indicator showed distinctively different patterns of spontaneous Ca2+ events in smooth muscle cells and ICC. Both cell types responded to carbachol by an increase in frequency of these events. In conclusion, guinea pig bladder detrusor ICC, both as isolated cells and within whole tissue preparations, respond to cholinergic stimulation by firing Ca2+ transients.
Original languageEnglish
Pages (from-to)F645-F655
Number of pages11
JournalAmerican Journal of Physiology - Renal Physiology
Volume294
Issue number3
DOIs
Publication statusPublished - Mar 2008

Fingerprint

Interstitial Cells of Cajal
Cholinergic Agents
Carbachol
Guinea Pigs
Urinary Bladder
Acetylcholine
Tetracaine
Ryanodine
Phospholipases
Nifedipine
Smooth Muscle Myocytes
Smooth Muscle
Swine

Keywords

  • confocal microscopy
  • c-Kit

Cite this

Johnston, L. ; Carson, C. ; Lyons, A.D. ; Davidson, R.A. ; McCloskey, K.D. / Cholinergic-induced Ca2+ signaling in interstitial cells of Cajal from the guinea pig bladder. In: American Journal of Physiology - Renal Physiology. 2008 ; Vol. 294, No. 3. pp. F645-F655.
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abstract = "Acetylcholine released from parasympathetic excitatory nerves activates contraction in detrusor smooth muscle. Immunohistochemical labeling of guinea pig detrusor with anti-c-Kit and anti-VAChT demonstrated a close structural relationship between interstitial cells of Cajal (ICC) and cholinergic nerves. The ability of guinea pig bladder detrusor ICC to respond to the acetylcholine analog, carbachol, was investigated in enzymatically dissociated cells, loaded with the Ca2+ indicator fluo 4AM. ICC fired Ca2+ transients in response to stimulation by carbachol (1/10 M). Their pharmacology was consistent with carbachol-induced contractions in strips of detrusor which were inhibited by 4-DAMP (1 M), an M3 receptor antagonist, but not by the M2 receptor antagonist methoctramine (1 M). The source of Ca2+ underlying the carbachol transients in isolated ICC was investigated using agents to interfere with influx or release from intracellular stores. Nifedipine (1 M) or Ni2 (30-100 M) to block Ca2+ channels or the removal of external Ca2 reduced the amplitude of the carbachol transients. Application of ryanodine (30 M) or tetracaine (100 M) abolished the transients. The phospholipase C inhibitor, U-73122 (2.5 M), significantly reduced the responses. 2-Aminoethoxydiethylborate (30 M) caused a significant reduction and Xestospongin C (1 M) was more effective, almost abolishing the responses. Intact in situ preparations of guinea pig bladder loaded with a Ca2+ indicator showed distinctively different patterns of spontaneous Ca2+ events in smooth muscle cells and ICC. Both cell types responded to carbachol by an increase in frequency of these events. In conclusion, guinea pig bladder detrusor ICC, both as isolated cells and within whole tissue preparations, respond to cholinergic stimulation by firing Ca2+ transients.",
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Cholinergic-induced Ca2+ signaling in interstitial cells of Cajal from the guinea pig bladder. / Johnston, L.; Carson, C.; Lyons, A.D.; Davidson, R.A.; McCloskey, K.D.

In: American Journal of Physiology - Renal Physiology, Vol. 294, No. 3, 03.2008, p. F645-F655.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cholinergic-induced Ca2+ signaling in interstitial cells of Cajal from the guinea pig bladder

AU - Johnston, L.

AU - Carson, C.

AU - Lyons, A.D.

AU - Davidson, R.A.

AU - McCloskey, K.D.

PY - 2008/3

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N2 - Acetylcholine released from parasympathetic excitatory nerves activates contraction in detrusor smooth muscle. Immunohistochemical labeling of guinea pig detrusor with anti-c-Kit and anti-VAChT demonstrated a close structural relationship between interstitial cells of Cajal (ICC) and cholinergic nerves. The ability of guinea pig bladder detrusor ICC to respond to the acetylcholine analog, carbachol, was investigated in enzymatically dissociated cells, loaded with the Ca2+ indicator fluo 4AM. ICC fired Ca2+ transients in response to stimulation by carbachol (1/10 M). Their pharmacology was consistent with carbachol-induced contractions in strips of detrusor which were inhibited by 4-DAMP (1 M), an M3 receptor antagonist, but not by the M2 receptor antagonist methoctramine (1 M). The source of Ca2+ underlying the carbachol transients in isolated ICC was investigated using agents to interfere with influx or release from intracellular stores. Nifedipine (1 M) or Ni2 (30-100 M) to block Ca2+ channels or the removal of external Ca2 reduced the amplitude of the carbachol transients. Application of ryanodine (30 M) or tetracaine (100 M) abolished the transients. The phospholipase C inhibitor, U-73122 (2.5 M), significantly reduced the responses. 2-Aminoethoxydiethylborate (30 M) caused a significant reduction and Xestospongin C (1 M) was more effective, almost abolishing the responses. Intact in situ preparations of guinea pig bladder loaded with a Ca2+ indicator showed distinctively different patterns of spontaneous Ca2+ events in smooth muscle cells and ICC. Both cell types responded to carbachol by an increase in frequency of these events. In conclusion, guinea pig bladder detrusor ICC, both as isolated cells and within whole tissue preparations, respond to cholinergic stimulation by firing Ca2+ transients.

AB - Acetylcholine released from parasympathetic excitatory nerves activates contraction in detrusor smooth muscle. Immunohistochemical labeling of guinea pig detrusor with anti-c-Kit and anti-VAChT demonstrated a close structural relationship between interstitial cells of Cajal (ICC) and cholinergic nerves. The ability of guinea pig bladder detrusor ICC to respond to the acetylcholine analog, carbachol, was investigated in enzymatically dissociated cells, loaded with the Ca2+ indicator fluo 4AM. ICC fired Ca2+ transients in response to stimulation by carbachol (1/10 M). Their pharmacology was consistent with carbachol-induced contractions in strips of detrusor which were inhibited by 4-DAMP (1 M), an M3 receptor antagonist, but not by the M2 receptor antagonist methoctramine (1 M). The source of Ca2+ underlying the carbachol transients in isolated ICC was investigated using agents to interfere with influx or release from intracellular stores. Nifedipine (1 M) or Ni2 (30-100 M) to block Ca2+ channels or the removal of external Ca2 reduced the amplitude of the carbachol transients. Application of ryanodine (30 M) or tetracaine (100 M) abolished the transients. The phospholipase C inhibitor, U-73122 (2.5 M), significantly reduced the responses. 2-Aminoethoxydiethylborate (30 M) caused a significant reduction and Xestospongin C (1 M) was more effective, almost abolishing the responses. Intact in situ preparations of guinea pig bladder loaded with a Ca2+ indicator showed distinctively different patterns of spontaneous Ca2+ events in smooth muscle cells and ICC. Both cell types responded to carbachol by an increase in frequency of these events. In conclusion, guinea pig bladder detrusor ICC, both as isolated cells and within whole tissue preparations, respond to cholinergic stimulation by firing Ca2+ transients.

KW - confocal microscopy

KW - c-Kit

U2 - 10.1152/ajprenal.00526.2007

DO - 10.1152/ajprenal.00526.2007

M3 - Article

VL - 294

SP - F645-F655

JO - American Journal of Physiology - Renal Physiology

JF - American Journal of Physiology - Renal Physiology

SN - 1931-857X

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