ISSN:
1432-2013
Keywords:
c-kit
;
Anti-c-kit-monoclonal antibody (ACK2)
;
Ca2+-Dependent Cl− current
;
Pacemaker activity
Source:
Springer Online Journal Archives 1860-2000
Topics:
Medicine
Notes:
Abstract 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 immuno-histochemically 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 voltageclamp at −40 mV, the majority of c -kif +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 Ca2+-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−(E Cl ) Moreover the rhythmic current was depressed by a Cl− channel blocker, 4-acetoamido-4-isothiocyanat-ostilbene-2,2′-disulphonic acid (SITS). The smooth muscle cells freshly dissociated from the same intestinal specimen revealed a Ca2+-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 Ca2+-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.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00410188
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