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Validation of a Mouse Model to Disrupt LINC Complexes in a Cell-specific Manner

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Defective ENaC processing and function in tissue kallikrein-deficient mice.

Nicolas Picard¹, Dominique Eladari, Soumaya El Moghrabi

¹INSERM U872 (Centre de Recherche des Cordeliers), Université Paris 5, Faculté de Médecine René Descartes, and Département de Physiologie, Hôpital Necker-Enfants Malades, F-75015, Paris, France.

The Journal of Biological Chemistry

|December 19, 2007

Summary

Tissue kallikrein (TK) activates epithelial sodium channels (ENaC) in the kidney and colon. This protease activity is crucial for regulating sodium transport and potentially blood pressure.

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Area of Science:

Physiology
Nephrology
Molecular Biology

Background:

Urinary tissue kallikrein (TK) excretion inversely correlates with blood pressure in humans and rodents.
The precise function of TK in the kidney, particularly its role in urinary fluid, remains largely unknown.
TK is synthesized in the kidney's connecting tubule and released into urine.

Purpose of the Study:

To investigate whether tissue kallikrein (TK) regulates apically expressed sodium transporters.
To determine the role of TK in renal and colonic sodium transport.
To elucidate the mechanism by which TK influences gamma-ENaC activity.

Main Methods:

Utilized gene-knockout mice lacking functional tissue kallikrein (TK-/-).
Employed semiquantitative immunoblotting to analyze protein expression in renal cortex.
Measured urinary sodium excretion, amiloride-sensitive potential difference, and intracellular sodium concentration in microperfused kidney tubules.

Main Results:

TK-/- mice exhibited an absence of the 70-kDa gamma-ENaC form and blunted urinary sodium excretion post-amiloride.
Amiloride-sensitive potential difference in the colon was decreased in TK-/- mice, but unaffected in the lungs.
In vitro studies showed TK promotes gamma-ENaC cleavage and increases intracellular sodium concentration in kidney tubules.

Conclusions:

Tissue kallikrein (TK) activates epithelial sodium channels (ENaC) in the kidney and colon.
TK's protease activity is responsible for processing and activating gamma-ENaC.
These findings suggest a novel role for TK in regulating sodium balance and blood pressure.