Characterization of three types of calcium channel in the luminal membrane of the distal nephron

We previously reported a dual kinetics of Ca2+ transport by the distal tubule luminal membrane of the kidney, suggesting the presence of several types of channels. To better characterize these channels, we examined the effects of specific inhibitors (i.e., diltiazem, an L-type channel; ω-conotoxin MVIIC, a P/Q-type channel; and mibefradil, a T-type channel antagonist) on 0.1 and 0.5 mM Ca2+ uptake by rabbit nephron luminal membranes. None of these inhibitors influenced Ca2+ uptake by the proximal tubule membranes. In contrast, in the absence of sodium (Na+), the three channel antagonists decreased Ca2+ transport by the distal membranes, and their action depended on the substrate concentrations: 50 µM diltiazem decreased 0.1 mM Ca2+ uptake from 0.65 ± 0.07 to 0.48 ± 0.06 pmol·µg–1·10 s–1 (P < 0.05) without influencing 0.5 mM Ca2+ transport, whereas 100 nM ω-conotoxin MVIIC decreased 0.5 mM Ca2+ uptake from 1.02 ± 0.05 to 0.90 ± 0.05 pmol·µg–1·10 s–1 (P < 0.02) and 1 µM mibefradil decreased it from 1.13 ± 0.09 to 0.94 ± 0.09 pmol·µg–1·10 s–1 (P < 0.05); the latter two inhibitors left 0.1 mM Ca2+ transport unchanged. Diltiazem decreased the Vmax of the high-affinity channels, whereas ω-conotoxin MVIIC and mibefradil influenced exclusively the Vmax of the low-affinity channels. These results not only confirm that the distal luminal membrane is the site of Ca2+ channels, but they suggest that these channels belong to the L, P/Q, and T types.Key words: renal calcium transport, calcium channels, diltiazem, mibefradil, ω-conotoxin.

Renal calcium transport: mechanisms and regulation--an overview

Renal calcium transport is described as the result of two processes, a paracellular, gradient-dependent process that predominates in most segments of the nephron and a transcellular, energy-dependent step that characterizes calcium transport in the distal convoluted tubule (DCT). Transcellular calcium transport involves entry into the DCT cell, possibly via channels, intracellular movement which appears to be facilitated by the presence of the vitamin D-dependent, cytosolic calcium-binding protein (CaBPr, calbindin D28k, mol mass approximately 28 kDa), and extrusion via the Ca-ATPase. Although much is known about calcium channels, their presence in renal tissue has only been demonstrated by preliminary studies. Quantitative data on CaBPr content of rat DCT are also unavailable, but theoretical analysis and early experimental values of intracellular self-diffusion of calcium have confirmed the need for an intracellular calcium "ferry," i.e., a molecule like CaBPr to amplify intracellular calcium movement. Available data on the plasma membrane Ca-ATPase are consistent with the extrusion kinetics attributed to the renal Ca-ATPase, but it has not been isolated, nor has its gene been cloned. Regulation and disorders of renal calcium transport are likely to involve one of the three transcellular steps, but indirect regulation by modification of the cell walls and molecules constituting the paracellular pathway cannot be excluded.

Renal Calcium Transport: Sites and Insights

In the kidney the absorption of calcium in the proximal tubule is mediated largely through passive transport processes, in the distal tubule by hormone-sensitive transcellular movement, whereas the loop on Henle represents a hybrid situation. The ability of calcium-transporting cells to accommodate wide ranges of transcellular movement without disturbing cytosolic calcium-dependent processes suggests the presence of mechanisms to coordinate apical entry and basolateral efflux of calcium.