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Mapping Intestinal Paracellular Perm Eability in Mice: Regional and Cellular Variability Under Physiological and

Mathilde Miquel1,2, Kadirey Verwaerde1, Anissa Edir-Kibri3

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Summary

This study reveals that intestinal permeability, or "leakiness," occurs around specific cells, and this can be detected using FM1-43FX (FM) dye. Chronic stress and lipid diets were found to increase this paracellular permeability.

Keywords:
chronic stressenteroendocrine cellsgoblet cellsintestinelipid absorptionparacellular permeabilitytuft cells

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Area of Science:

  • Gastroenterology
  • Cell Biology
  • Physiology

Background:

  • Intestinal paracellular permeability is crucial for homeostasis.
  • Understanding the pathways and regulation of intestinal permeability is vital for diagnosing and treating gastrointestinal disorders.

Purpose of the Study:

  • To analyze intestinal paracellular permeability ex vivo using fluorescent dyes.
  • To investigate the impact of chronic stress and lipid diets on intestinal permeability.
  • To validate the FM1-43FX (FM) methodology for detecting increased paracellular permeability.

Main Methods:

  • Incubation of mouse intestinal tissue segments at 0°C with fluorescent dyes FM1-43FX (FM) or TRITC-dextran 3 kDa lysine-fixable (TD3L).
  • Confocal microscopy was used to visualize dye penetration patterns.
  • Analysis was performed on healthy mice, mice under chronic stress (glucocorticoid exposure), and mice fed lipid diets.

Main Results:

  • In healthy mice, FM and TD3L dyes showed restricted but specific penetration around certain cell types (Goblet, enteroendocrine, tuft, apoptotic cells) in the small intestine and colon.
  • This penetration pattern at 0°C indicates a paracellular 'leak' pathway, distinct from transcellular endocytosis.
  • Chronic stress increased FITC-dextran 4 kDa permeability in vivo and FM-detected paracellular permeability ex vivo in the colon.
  • A lipid-rich meal increased ex vivo permeability around jejunal enterocytes.

Conclusions:

  • The study identifies specific paracellular pathways around specialized intestinal cells involved in luminal sensing.
  • The FM1-43FX (FM) dye methodology is effective in detecting and localizing increased paracellular permeability ex vivo.
  • Pathophysiological conditions like chronic stress and lipid-rich diets can significantly increase intestinal paracellular permeability.