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Fluid absorption in isolated perfused colonic crypts

S K Singh1, H J Binder, W F Boron

  • 1Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

The Journal of Clinical Investigation
|November 1, 1995
PubMed
Summary
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Isolated rat colon crypts absorb fluid in their basal state. Neurohumoral stimulation, however, induces net fluid secretion, challenging the traditional model of intestinal ion transport.

Area of Science:

  • Gastroenterology
  • Physiology
  • Cell Biology

Background:

  • The established model posits distinct absorptive (surface cells) and secretory (crypt cells) functions in the intestine.
  • Direct measurement of fluid movement within intestinal crypts is essential for validating this model.

Purpose of the Study:

  • To adapt renal tubule microperfusion techniques for dissecting and perfusing isolated rat colon crypts.
  • To directly measure fluid movement in single, isolated crypts to understand their transport functions.

Main Methods:

  • Hand-dissection and microperfusion of single, isolated crypts from rat distal colon.
  • Morphologic analysis to ensure crypt purity and integrity.
  • Measurement of net fluid movement under basal conditions and after stimulation with neurohumoral agonists.

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Main Results:

  • Basal state: Isolated crypts demonstrated net fluid absorption (0.34 +/- 0.01 nl.mm-1.min-1), dependent on sodium and bicarbonate.
  • Stimulated state: Addition of dibutyryl-cyclic AMP, vasoactive intestinal peptide, or acetylcholine induced reversible net fluid secretion (-0.35 +/- 0.01 nl.mm-1.min-1).

Conclusions:

  • Colonic crypt cells possess constitutive absorptive capabilities.
  • Fluid secretion in crypt cells is regulated by neurohumoral agonists, likely released from lamina propria cells.
  • This study establishes a functional model of the intact colon crypt for future mechanistic investigations of fluid and electrolyte transport.