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Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity.
McKinley, Eliot T; Sui, Yunxia; Al-Kofahi, Yousef; Millis, Bryan A; Tyska, Matthew J; Roland, Joseph T; Santamaria-Pang, Alberto; Ohland, Christina L; Jobin, Christian; Franklin, Jeffrey L; Lau, Ken S; Gerdes, Michael J; Coffey, Robert J.
Afiliação
  • McKinley ET; Epithelial Biology Center and.
  • Sui Y; Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
  • Al-Kofahi Y; General Electric Global Research Center, Niskayuna, New York, USA.
  • Millis BA; General Electric Global Research Center, Niskayuna, New York, USA.
  • Tyska MJ; Department of Cell and Developmental Biology.
  • Roland JT; Cell Imaging Shared Resource, and.
  • Santamaria-Pang A; Epithelial Biology Center and.
  • Ohland CL; Department of Cell and Developmental Biology.
  • Jobin C; Epithelial Biology Center and.
  • Franklin JL; Department of Surgery, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
  • Lau KS; General Electric Global Research Center, Niskayuna, New York, USA.
  • Gerdes MJ; Department of Medicine.
  • Coffey RJ; Department of Medicine.
JCI Insight ; 2(11)2017 Jun 02.
Article em En | MEDLINE | ID: mdl-28570279
ABSTRACT
Intestinal tuft cells are a rare, poorly understood cell type recently shown to be a critical mediator of type 2 immune response to helminth infection. Here, we present advances in segmentation algorithms and analytical tools for multiplex immunofluorescence (MxIF), a platform that enables iterative staining of over 60 antibodies on a single tissue section. These refinements have enabled a comprehensive analysis of tuft cell number, distribution, and protein expression profiles as a function of anatomical location and physiological perturbations. Based solely on DCLK1 immunoreactivity, tuft cell numbers were similar throughout the mouse small intestine and colon. However, multiple subsets of tuft cells were uncovered when protein coexpression signatures were examined, including two new intestinal tuft cell markers, Hopx and EGFR phosphotyrosine 1068. Furthermore, we identified dynamic changes in tuft cell number, composition, and protein expression associated with fasting and refeeding and after introduction of microbiota to germ-free mice. These studies provide a foundational framework for future studies of intestinal tuft cell regulation and demonstrate the utility of our improved MxIF computational methods and workflow for understanding cellular heterogeneity in complex tissues in normal and disease states.
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Texto completo: 1 Base de dados: MEDLINE Idioma: En Revista: JCI Insight Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Idioma: En Revista: JCI Insight Ano de publicação: 2017 Tipo de documento: Article