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1 Departments of Medicine and Physiology, Division of Nephrology, University of Alabama at Birmingham, Birmingham, AL, USA; Institute of Pathophysiology, Hungarian Academy of Sciences and Semmelweis University Nephrolgy Research Group, Budapest, Hungary
2 Departments of Medicine and Physiology, Division of Nephrology, University of Alabama at Birmingham, Birmingham, AL, USA
3 Institute of Pathophysiology, Hungarian Academy of Sciences and Semmelweis University Nephrolgy Research Group, Budapest, Hungary
* To whom correspondence should be addressed. E-mail: pkomlosi{at}uab.edu.
One component of the macula densa (MD) tubuloglomerular feedback (TGF) signaling pathway may involve basolateral release of ATP through a maxi-anion channel. Release of ATP has previously been studied during a maximal luminal sodium chloride ([NaCl]L) stimulus (20 to 150 mmol/L, NaCl). Whether MD ATP release occurs during changes in [NaCl]L within the physiological range (20 to 60 mmol/L, NaCl) has not been examined. Also, since TGF is known to be enhanced by low dietary salt intake we examined the pattern of macula densa-ATP release from salt restricted rabbits. ATP release from the isolated perfused thick ascending limb-MD was assessed using fluorescence microscopy and fura-2-loaded cultured mouse mesangial cells as biosensors. The mesangial-biosensor cells, which contain purinergic receptors and produce elevated [Ca2+]i upon ATP binding, were placed next to the MD basolateral membrane. Elevations in [NaCl]L between 0 and 80 mmol/L, in 20 mmol/L increments, caused step-wise increases in [Ca2+]i with the highest increase observed at ~ 60 mmol/L [NaCl]L. 10-4 mol/L luminal furosemide blocked ATP release, which suggests that the efflux of ATP required MD Na:2Cl:K cotransport. Low-salt diet for one week increased the magnitude of [NaCl]L-dependent elevations in biosensor [Ca2+]i by 2-fold, while high-salt intake had no effect. In summary, ATP release occurs over the same range (20 to 60 mmol/L) of [NaCl]L as previously reported for TGF responses and, like TGF, ATP release was enhanced by dietary salt restriction. Thus, these two findings are consistent with the role of MD ATP release as a signaling component of the tubuloglomerular feedback pathway.
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