Action potential generation, Kit receptor immunohistochemistry and morphology of steel-Dickie (Sl/Sld) mutant mouse small intestine.
In contrast to wild-type mice, homozygotes with mutations of the W locus do not express the functional Kit receptor and are severely deficient in the Auerbach's plexus (AP)-associated subtype of interstitial cells of Cajal (ICC-AP). With a morphologically intact neural and muscular structure, the absence in these mutants of both small-intestinal slow waves and ICC-AP constitutes strong evidence for a key role of ICC-AP as pacemaker cells. In steel-Dickie mutant mice (Sl/Sld), the gene coding for the Kit ligand (stem cell factor) is defective. We examined Sl/Sld mutants and controls with intracellular microelectrode techniques, combined with light and electron microscopy. The absence of the normal Kit ligand (Sl/Sld mice) had very similar effects as the absence of the Kit receptor in viable mice, mutated at the White spotting, W, locus (W/Wv mice), in that neither slow waves, nor Kit receptor immunoreactivity in the region of Auerbach's plexus nor ICC-AP were present in the small intestine. In the Sl/Sld mouse, the smooth muscle cells generated action potentials at variable frequencies from a depolarized cell membrane of -40 to -55 mV. Increasing excitability by K channel blockers created many different patterns of action potential generation and the frequency increased from approximately 16 cpm to 66 cpm. This was in sharp contrast to control mice where action potentials were always restricted to the plateau phase of the slow waves and the slow wave frequency remained constant at approximately 39 cpm. Our data provide further strong support for the identification of ICC-AP as small-intestinal pacemaker cells. In addition, they provide a basis for the understanding of intestinal motor function without pacemaker activity.
Mikkelsen
H B
HB
Institute of Medical Anatomy, University of Copenhagen, Denmark. hmikkelsen@mai.ku.dk
Malysz
J
J
Huizinga
J D
JD
Thuneberg
L
L
D000200:Action Potentials / Q000502:physiology; D000818:Animals; D003911:Dextrans; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D007150:Immunohistochemistry; D007421:Intestine, Small / Q000378:metabolism* / Q000502:physiology* / Q000648:ultrastructure; D008564:Membrane Potentials / Q000502:physiology; D008751:Methylene Blue; D051379:Mice; D008818:Mice, Neurologic Mutants; D008854:Microscopy, Electron; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics / Q000378:metabolism*; D012996:Solutions
D003911:Dextrans; D012996:Solutions; C015219:fluorescein isothiocyanate dextran; D019009:Proto-Oncogene Proteins c-kit; D016650:Fluorescein-5-isothiocyanate; D008751:Methylene Blue