L-arginine effects on Na+ transport in M-1 mouse cortical collecting duct cells--a cationic amino acid absorbing epithelium.

Journal article

The effect of L-arginine on transepithelial ion transport was examined in cultured M-1 mouse renal cortical collecting duct (CCD) cells using continuous short circuit current (Isc) measurements in HCO3-/CO2 buffered solution. Steady state Isc averaged 73.8 +/- 3.2 microA/cm2 (n = 126) and was reduced by 94 +/- 0.6% (n = 16) by the apical addition of 100 microM amiloride. This confirms that the predominant electrogenic ion transport in M-1 cells is Na+ absorption via the epithelial sodium channel (ENaC). Experiments using the cationic amino acid L-lysine (radiolabeled) as a stable arginine analogue show that the combined activity of an apical system y+ and a basal amino acid transport system y+L are responsible for most cationic amino acid transport across M-1 cells. Together they generate net absorptive cationic amino acid flux. Application of L-arginine (10 mM) either apically or basolaterally induced a transient peak increase in Isc averaging 36.6 +/- 5.4 microA/cm2 (n = 19) and 32.0 +/- 7.2 microA/cm2 (n = 8), respectively. The response was preserved in the absence of bath Cl- (n = 4), but was abolished either in the absence of apical Na+ (n = 4) or by apical addition of 100 microM amiloride (n = 6). L-lysine, which cannot serve as a precursor of NO, caused a response similar to that of L-arginine (n = 4); neither L-NMMA (100 microM; n = 3) nor L-NAME (1 mM; n = 4) (both NO-synthase inhibitors) affected the Isc response to L-arginine. The effects of arginine or lysine were replicated by alkalinization that mimicked the transient alkalinization of the bath solution upon addition of these amino acids. We conclude that in M-1 cells L-arginine stimulates Na+ absorption via a pH-dependent, but NO-independent mechanism. The observed net cationic amino acid absorption will counteract passive cationic amino acid leak into the CCD in the presence of electrogenic Na+ transport, consistent with reports of stimulated expression of Na+ and cationic amino acid transporters by aldosterone.

Authors: Howard, D; Cuffe, J; Boyd, C; Korbmacher, C

• Journal: Journal of membrane biology
• Volume: 180
• Issue: 2
• Pages: 111-121
• Publication date: 2001-03-01
• DOI: 10.1007/s002320010063
• EISSN: 1432-1424
• ISSN: 0022-2631
• Language: English
• Keywords: Urothelium; Cell Line; Histidine; Mice; Diuretics; Electrophysiology; Arginine; Sodium; S-Nitroso-N-Acetylpenicillamine; Lysine; Kidney Tubules, Collecting; Amiloride; Penicillamine; Cell Polarity; omega-N-Methylarginine; Enzyme Inhibitors; Animals; Epithelial Cells; Ion Transport