Cytosolic [Ca2+] signaling pathway in macula densa cells.
Previous micropuncture studies suggested that macula densa (MD) cells might detect variations in luminal sodium chloride concentration ([NaCl]l) through changes in cytosolic calcium ([Ca2+]c). To test this hypothesis, MD [Ca2+]c was measured with fluorescence microscopy using fura 2 in the isolated perfused thick ascending limb with attached glomerulus preparation dissected from rabbit kidney. Tubules were bathed and perfused with a Ringer solution, [NaCl]l was varied and isosmotically replaced with N-methyl-D-glucamine cyclamate. Control [Ca2+]c, during perfusion with 25 mM NaCl and 150 mM NaCl in the bath, averaged 101. 6 +/- 8.2 nM (n = 21). Increasing [NaCl]l to 150 mM elevated [Ca2+]c by 39.1 +/- 5.2 nM (n = 21, P < 0.01). This effect was concentration dependent between zero and 60 mM [NaCl]l. The presence of either luminal furosemide or basolateral nifedipine or 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), a potent Cl- channel blocker, significantly reduced resting [Ca2+]c and abolished the increase in [Ca2+]c in response to increased [NaCl]l. Nifedipine failed to produce a similar inhibitory effect when added exclusively to the luminal perfusate. Also, 100 nM BAY K 8644, a voltage-gated Ca2+ channel agonist, added to the bathing solution increased [Ca2+]c by 33.2 +/- 8.1 nM (n = 5, P < 0.05). These observations suggest that MD cells may detect variations in [NaCl]l through a signaling pathway that includes Na+-2Cl--K+ cotransport, basolateral membrane depolarization via Cl- channels, and Ca2+ entry through voltage-gated Ca2+ channels.
Peti-Peterdi
J
J
Nephrology Research and Training Center, Division of Nephrology, Departments of Medicine and Physiology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Bell
P D
PD
D000818:Animals; D002118:Calcium / Q000502:physiology*; D003600:Cytosol / Q000502:physiology; D005456:Fluorescent Dyes; D016257:Fura-2; D008138:Loop of Henle / Q000502:physiology*; D011817:Rabbits; D015398:Signal Transduction; D012965:Sodium Chloride / Q000378:metabolism / Q000494:pharmacology
D005456:Fluorescent Dyes; D012965:Sodium Chloride; D002118:Calcium; D016257:Fura-2