Document Detail


Rhythmic Cl- current and physiological roles of the intestinal c-kit-positive cells.
MedLine Citation:
PMID:  9026776     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chronic injection of an anti-c-KIT receptor tyrosine kinase monoclonal antibody (ACK2) results in the disruption of the normal motility patterns of young BALB/c mice intestine. This effect is accompanied by a drastic decrease in the number of intestinal c-kit-expressing (c-kit+) cells when studied immunohistochemically with the fluorescence-labelled antibody. In order to clarify the mechanism underlying the ACK2 action and the physiological roles of intestinal c-kit+ cells, we studied the excitability of intestinal c-kit+ cells in primary culture by use of the nystatin perforated-patch-clamp technique. Under voltage-clamp at -40 mV, the majority of c-kit+ cells tested (59/70) elicited rhythmic current waves with an amplitude and frequency of 263 +/- 24 pA and 2.30 +/- 0.25 cycles/min (mean +/- SEM), respectively. Intracellular perfusion of the c-kit+ cells with ethylenebis (okonitrilo) tetraacetate (EGTA) as well as a nominally Ca(2+)-free external solution or low holding voltage (< -60 mV) prevented the rhythmic current. The reversal potential of the rhythmic current was close to the equilibrium potential for Cl-(ECl). Moreover the rhythmic current was depressed by a Cl- channel blocker, 4-acetoamido-4-isothiocyanat-ostilbene-2,2'-disulphoni c acid (SITS). The smooth muscle cells freshly dissociated from the same intestinal specimen revealed a Ca(2+)-activated K+ current, as has been described in a variety of smooth muscle cells. Cultured smooth muscle cells from the ileum preparation lacked neither the Ca(2+)-activated K+ nor rhythmic Cl- currents. Smooth muscle cells freshly dissociated from the same ileum preparation and those in culture showed no immunoreactivity with the labelled ACK2, which was consistent with our previous in situ study. Results provided direct evidence that the intestinal c-kit+ cells, but not the smooth muscle cells, possess a rhythmic Cl- current oscillation, suggesting their participation in pacemaker activity for the peristaltic gut movement.
Authors:
N Tokutomi; H Maeda; Y Tokutomi; D Sato; M Sugita; S Nishikawa; S Nishikawa; J Nakao; T Imamura; K Nishi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Pflügers Archiv : European journal of physiology     Volume:  431     ISSN:  0031-6768     ISO Abbreviation:  Pflugers Arch.     Publication Date:  1995 Dec 
Date Detail:
Created Date:  1997-02-18     Completed Date:  1997-02-18     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0154720     Medline TA:  Pflugers Arch     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  169-77     Citation Subset:  IM    
Affiliation:
Department of Pharmacology, Kumamoto University School of Medicine, Japan.
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MeSH Terms
Descriptor/Qualifier:
4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / pharmacology
Animals
Animals, Newborn
Antibodies, Monoclonal / diagnostic use
Chelating Agents / pharmacology
Chloride Channels / drug effects,  metabolism*
Egtazic Acid / pharmacology
Electrophysiology
Fluorescent Antibody Technique, Indirect
Immunohistochemistry
Intestines / cytology*,  drug effects,  enzymology*
Male
Mice
Mice, Inbred BALB C
Muscle Contraction / drug effects
Muscle, Smooth / cytology*,  drug effects,  enzymology*
Nystatin / diagnostic use
Patch-Clamp Techniques
Receptor Protein-Tyrosine Kinases / antagonists & inhibitors,  metabolism*
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Chelating Agents; 0/Chloride Channels; 1400-61-9/Nystatin; 27816-59-7/4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; 67-42-5/Egtazic Acid; EC 2.7.10.1/Receptor Protein-Tyrosine Kinases

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