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Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts
Published on: February 6, 2021
Membrane properties of rat colonic crypts during early postnatal development
1Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
Insights
Potassium conductance (GK) decreases during rat colonic crypt maturation. Immature crypts show higher GK, with heterogeneous distribution along the crypt-surface axis, indicating developmental changes in ion transport.
Area of Science:
- Physiology
- Gastroenterology
- Cell Biology
Background:
- Plasma membrane transporters and channels change during development.
- Intestinal crypt maturation involves significant cellular changes.
- Knowledge of cell membrane conductances during colonic crypt development is limited.
Purpose of the Study:
- To investigate developmental changes in potassium conductance (GK) in rat colonic crypts.
- To examine the heterogeneity of GK along the crypt-surface axis during maturation.
Main Methods:
- Whole-cell patch clamp technique.
- Investigation of rat colonic crypts at different developmental stages.
- Assessment of ion channel activity using specific blockers (Ba2+, TEA).
Main Results:
- Immature crypts exhibited higher resting membrane potential and outward currents compared to mature crypts.
- Potassium conductance (GK) was higher in immature crypts and reduced by Ba2+ and tetraethylammonium (TEA).
- TEA-sensitive conductance was heterogeneously distributed along the crypt-surface axis, higher in undifferentiated crypt base cells.
Conclusions:
- Colonocyte differentiation and colonic crypt maturation are associated with a decrease in GK.
- Axial heterogeneity of GK is present early in postnatal life during rapid colonic epithelial growth.
Abstract:
The expression patterns of plasma membrane transporters and channels undergo developmental changes during cell differentiation and postnatal development. Until now, little is known about cell membrane conductances during maturation of intestinal crypts even though it is accepted that the crypts undergo dramatic changes during this period of life. Using the whole-cell patch clamp technique, we investigated the developmental changes of potassium conductance (GK) in rat colonic crypts and its heterogeneity along the crypt-surface axis. The resting membrane potential of immature crypts was sensitive to Ba2+ and was higher than in mature crypts. The whole cell conductance during outward currents was higher in immature crypts than in the crypts of adult animals and was reduced by Ba2+ and tetraethylammonium (TEA). With Na+-containing Ringer solution in the bath and K+-containing solution in the pipette, Ba2+- and TEA-sensitive outward currents and conductances were higher in immature than in mature crypts. The TEA-sensitive conductance in immature crypts was distributed heterogenously along the crypt-surface axis with higher values in undifferentiated cells of the crypt base than in differentiated cells of the crypt surface. These data indicate that (1) both colonocyte differentiation and maturation of colonic crypt epithelium are associated with decreasing GK and (2) the axial heterogeneity already exists early in postnatal life when the colonic epithelium undergoes rapid proliferation and growth associated with elongation and bifurcation of the crypts.

