Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine

2007 ◽  
Vol 292 (1) ◽  
pp. C497-C507 ◽  
Author(s):  
Hui Chen ◽  
Doug Redelman ◽  
Seungil Ro ◽  
Sean M. Ward ◽  
Tamás Ördög ◽  
...  
Keyword(s):  
Small Intestine ◽  
Substance P ◽  
Interstitial Cells Of Cajal ◽  
Soluble Guanylate Cyclase ◽  
Fluorescent Microscopy ◽  
Circular Muscle ◽  
Interstitial Cells ◽  
Functional Classes ◽  
Neurokinin 1 ◽  
Cells Of Cajal

Specific functions of interstitial cells of Cajal (ICC) have been linked to distinct classes that differ by morphology and distribution. In the small intestine, slow wave-generating ICC are located in the myenteric region (ICC-MY), whereas ICC that mediate neuromuscular neurotransmission occur either throughout the circular muscle layer (intramuscular ICC, ICC-IM) or in association with the deep muscular plexus (ICC-DMP). Selective isolation of ICC to characterize specific properties has been difficult. Recently, neurokinin-1 receptors have been detected in murine ICC-DMP and neurons but not in ICC-MY. Here we identified and isolated ICC-DMP/IM by receptor-mediated internalization of fluorescent substance P and Kit immunofluorescence. Specificity of labeling was verified by confocal microscopy. Mouse and human ICC-DMP/IM were detected in suspension by fluorescent microscopy and harvested for RT-PCR with micropipettes. The isolated cells expressed Kit but not markers for neurons, smooth muscle, or antigen-presenting cells. ICC-DMP expressed neurokinin-1 receptor, M2 and M3 muscarinic receptors, P2Y1 and P2Y4 purinergic receptors, VIP receptor 2, soluble guanylate cyclase-1 subunits, and protein kinase G. L- or T-type Ca2+ channels were not detected in these cells. ICC-MY and ICC-DMP were simultaneously detected and enumerated by flow cytometry and sorted to purity by fluorescence-activated cell sorting. In summary, functional classes of ICC have distinct molecular identities that can be used to selectively identify and harvest these cells with, for example, receptor-mediated uptake of substance P and Kit immunofluorescence. ICC-DMP express neurotransmitter receptors and signaling intermediate molecules that are consistent with their role in neuromuscular neurotransmission.

Activation of neurokinin 1 receptors on interstitial cells of Cajal of the guinea-pig small intestine by substance P

1998 ◽  
Vol 110 (3) ◽  
pp. 263-271 ◽  
Author(s):  
S. T. Lavin ◽  
B. R. Southwell ◽  
R. Murphy ◽  
K. M. Jenkinson ◽  
J. B. Furness
Keyword(s):  
Small Intestine ◽  
Substance P ◽  
Guinea Pig ◽  
Interstitial Cells Of Cajal ◽  
Interstitial Cells ◽  
Neurokinin 1 ◽  
Cells Of Cajal

Morphology of the canine pyloric sphincter in relation to function

1989 ◽  
Vol 67 (12) ◽  
pp. 1560-1573 ◽  
Author(s):  
E. E. Daniel ◽  
I. Berezin ◽  
H. D. Allescher ◽  
H. Manaka ◽  
V. Posey-Daniel
Keyword(s):  
Smooth Muscle ◽  
Substance P ◽  
Smooth Muscle Cells ◽  
Gap Junctions ◽  
Vasoactive Intestinal Polypeptide ◽  
Interstitial Cells Of Cajal ◽  
Circular Muscle ◽  
Interstitial Cells ◽  
Pyloric Sphincter ◽  
Cells Of Cajal

The ultrastructure and immunocytochemistry of the canine distal pyloric muscle loop, the pyloric sphincter, were studied. Cells in this muscle were connected by gap junctions, fewer than in the antrum or corpus. The sphincter had a dense innervation and a sparse population of interstitial cells of Cajal. Most such cells were of the circular muscle type but a few were of the type in the myenteric plexus. Nerves were sometimes associated with interstitial cell profiles, but most nerves were neither close to nor associated with interstitial cells nor close to smooth muscle cells. Nerve profiles were characterized by an unusually high proportion of varicosities with a majority or a high proportion of large granular vesicles. Many of these were shown to contain material immunoreactive for vasoactive intestinal polypeptide (VIP) and some had substance P (SP) immunoreactive material. All were presumed to be peptidergic. VIP was present in a higher concentration in this muscle than in adjacent antral or duodenal circular muscle. Interstitial cells of Cajal made gap junctions to smooth muscle and to one another and might provide myogenic pacemaking activity for this muscle, but there was no evidence of a close or special relationship between nerves with VIP or SP and these cells. The absence of close relationships between nerves and either interstitial cells or smooth muscle cells leaves unanswered questions about the structural basis for previous observations of discrete excitatory responses or pyloric sphincter to single stimuli or nerves up to one per second. In conclusion, the structural observations suggest that this muscle has special neural and myogenic control systems and that interstitial cells may function to control myogenic activity of this muscle but not to mediate neural signals.Key words: vasoactive intestinal polypeptide, interstitial cells of Cajal, neuropeptides, gap junctions, substance P.

Differential gene expression in functional classes of interstitial cells of Cajal in murine small intestine

2007 ◽  
Vol 31 (3) ◽  
pp. 492-509 ◽  
Author(s):  
Hui Chen ◽  
Tamás Ördög ◽  
Junwei Chen ◽  
David L. Young ◽  
Michael R. Bardsley ◽  
...  
Keyword(s):  
Gene Expression ◽  
Small Intestine ◽  
Motor Neurons ◽  
Interstitial Cells Of Cajal ◽  
Interstitial Cells ◽  
Cell Junction ◽  
Process Guidance ◽  
Functional Classes ◽  
Murine Small Intestine ◽  
Cells Of Cajal

Interstitial cells of Cajal (ICC) have important functions in regulation of motor activity in the gastrointestinal tract. In murine small intestine, ICC are gathered in the regions of the myenteric plexus (ICC-MY) and the deep muscular plexus (ICC-DMP). These two classes of ICC have different physiological functions. ICC-MY are pacemaker cells and generate the slow-wave electrical rhythmicity of gastrointestinal organs. ICC-DMP form synaptic connections with the varicose nerve terminals of enteric motor neurons and are involved in reception and transduction of motor neurotransmission. Gene expression underlying specific functions of ICC classes is incompletely understood. In the present study, we used recently developed highly selective techniques to isolate the two functional ICC classes from enzymatically dispersed intestinal muscles by fluorescence-activated cell sorting. The transcriptomes of ICC-MY and ICC-DMP were investigated using oligonucleotide microarray analysis. Differential expression of functional groups of genes defined by standard gene ontology terms was also studied. There were substantial numbers of genes expressed more abundantly in ICC than in the tunica muscularis, and we also detected marked phenotypic differences between ICC-MY and ICC-DMP. Notably, genes related to cell junction, process guidance, and vesicle trafficking were upregulated in ICC. Consistent with their specific functions, metabolic and Ca2+ transport genes were relatively upregulated in ICC-MY, whereas genes for signaling proteins involved in transduction of neurotransmitter functions were relatively upregulated in ICC-DMP. Our results may lead to the identification of novel biomarkers for ICC and provide directions for further studies designed to understand ICC function in health and disease.

Immunohistochemical Evidence for Subclasses of Interstitial Cells of Cajal Associated with the Deep Muscular Plexus in Rat Small Intestine

1997 ◽  
Vol 3 (S2) ◽  
pp. 255-256
Author(s):  
A.M.R Yunker ◽  
K.A. Roth
Keyword(s):  
Small Intestine ◽  
Interstitial Cells Of Cajal ◽  
Longitudinal Muscle ◽  
Neuron Specific Enolase ◽  
Circular Muscle ◽  
Interstitial Cells ◽  
Nerve Fibers ◽  
Male Rats ◽  
Confocal Laser ◽  
Cells Of Cajal

Interstitial cells of Cajal (ICCs) were first described as primitive neurons found within organs innervated by autonomie nerves. In the gut, ICCs are juxtaposed among enteric nerve fibers and smooth muscle cells, suggesting they may modulate enteric neurotransmission and affect motility. The recent discovery of neurokinin-1 receptor (NKrl)-like immunoreactivity (ir) on ICCs has strengthened this hypothesis. This study compared the distribution of NKlr-ir to the staining patterns of other reported markers of ICCs including cholera toxin subunit b (CTB), neuron specific enolase (NSE), NADH diaphorase, NADPH diaphorase, nitric oxide synthase (NOS), and vimentin. Albino male rats were anesthetized and whole mount preparations of myenteric plexus-longitudinal muscle with attached circular muscle were dissected. ICCs were stained using multi-label techniques and cells were visualized with a Sarastro 2000 confocal laser scanning microscope equipped with Image Space software.In the small intestine, ICCs associated with circular muscle, but not longitudinal muscle, expressed NKlr-ir.

Ultrastructure of interstitial cells of Cajal in circular muscle of human small intestine

1993 ◽  
Vol 104 (2) ◽  
pp. 343-350 ◽  
Author(s):  
Jüri Johannes Rumessen ◽  
Hanne Birte Mikkelsen ◽  
Klaus Qvortrup ◽  
Lars Thuneberg
Keyword(s):  
Small Intestine ◽  
Interstitial Cells Of Cajal ◽  
Circular Muscle ◽  
Interstitial Cells ◽  
Human Small Intestine ◽  
Cells Of Cajal

scholarly journals A myogenic motor pattern in mice lacking myenteric interstitial cells of Cajal explained by a second coupled oscillator network

2020 ◽  
Vol 318 (2) ◽  
pp. G225-G243 ◽  
Author(s):  
Sean P. Parsons ◽  
Jan D. Huizinga
Keyword(s):  
Mathematical Model ◽  
Small Intestine ◽  
Interstitial Cells Of Cajal ◽  
Motor Pattern ◽  
Circular Muscle ◽  
Interstitial Cells ◽  
Mutant Mice ◽  
Coupled Oscillator ◽  
Oscillator Network ◽  
Cells Of Cajal

The interstitial cells of Cajal associated with the myenteric plexus (ICC-MP) are a network of coupled oscillators in the small intestine that generate rhythmic electrical phase waves leading to corresponding waves of contraction, yet rhythmic action potentials and intercellular calcium waves have been recorded from c-kit-mutant mice that lack the ICC-MP, suggesting that there may be a second pacemaker network. The gap junction blocker carbenoxolone induced a “pinstripe” motor pattern consisting of rhythmic “stripes” of contraction that appeared simultaneously across the intestine with a period of ~4 s. The infinite velocity of these stripes suggested they were generated by a coupled oscillator network, which we call X. In c-kit mutants rhythmic contraction waves with the period of X traveled the length of the intestine, before the induction of the pinstripe pattern by carbenoxolone. Thus X is not the ICC-MP and appears to operate under physiological conditions, a fact that could explain the viability of these mice. Individual stripes consisted of a complex pattern of bands of contraction and distension, and between stripes there could be slide waves and v waves of contraction. We hypothesized that these phenomena result from an interaction between X and the circular muscle that acts as a damped oscillator. A mathematical model of two chains of coupled Fitzhugh–Nagumo systems, representing X and circular muscle, supported this hypothesis. The presence of a second coupled oscillator network in the small intestine underlines the complexity of motor pattern generation in the gut. NEW & NOTEWORTHY Physiological experiments and a mathematical model indicate a coupled oscillator network in the small intestine in addition to the c-kit-expressing myenteric interstitial cells of Cajal. This network interacts with the circular muscle, which itself acts as a system of damped oscillators, to generate physiological contraction waves in c-kit (W) mutant mice.

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