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

Publication ,  Journal Article
Chen, H; Redelman, D; Ro, S; Ward, SM; Ördög, T; Sanders, KM
Published in: American Journal of Physiology-Cell Physiology
January 2007

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, M and M muscarinic receptors, P2Y and P2Y purinergic receptors, VIP receptor 2, soluble guanylate cyclase-1 subunits, and protein kinase G. L- or T-type Ca 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.

Duke Scholars

Published In

American Journal of Physiology-Cell Physiology

DOI

EISSN

1522-1563

ISSN

0363-6143

Publication Date

January 2007

Volume

292

Issue

1

Start / End Page

C497 / C507

Publisher

American Physiological Society

Related Subject Headings

  • Physiology
  • 1116 Medical Physiology
  • 0606 Physiology
  • 0601 Biochemistry and Cell Biology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Chen, H., Redelman, D., Ro, S., Ward, S. M., Ördög, T., & Sanders, K. M. (2007). Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine. American Journal of Physiology-Cell Physiology, 292(1), C497–C507. https://doi.org/10.1152/ajpcell.00147.2006
Chen, Hui, Doug Redelman, Seungil Ro, Sean M. Ward, Tamás Ördög, and Kenton M. Sanders. “Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine.” American Journal of Physiology-Cell Physiology 292, no. 1 (January 2007): C497–507. https://doi.org/10.1152/ajpcell.00147.2006.
Chen H, Redelman D, Ro S, Ward SM, Ördög T, Sanders KM. Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine. American Journal of Physiology-Cell Physiology. 2007 Jan;292(1):C497–507.
Chen, Hui, et al. “Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine.” American Journal of Physiology-Cell Physiology, vol. 292, no. 1, American Physiological Society, Jan. 2007, pp. C497–507. Crossref, doi:10.1152/ajpcell.00147.2006.
Chen H, Redelman D, Ro S, Ward SM, Ördög T, Sanders KM. Selective labeling and isolation of functional classes of interstitial cells of Cajal of human and murine small intestine. American Journal of Physiology-Cell Physiology. American Physiological Society; 2007 Jan;292(1):C497–C507.

Published In

American Journal of Physiology-Cell Physiology

DOI

EISSN

1522-1563

ISSN

0363-6143

Publication Date

January 2007

Volume

292

Issue

1

Start / End Page

C497 / C507

Publisher

American Physiological Society

Related Subject Headings

  • Physiology
  • 1116 Medical Physiology
  • 0606 Physiology
  • 0601 Biochemistry and Cell Biology