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We dissected and perfused in vitro rabbit cortical thick ascending limbs (CTAL) and distal convoluted tubule with the glomerulus attached intact. Basolateral membrane voltage (Vbl) and intracellular Ca2+ concentration ([Ca2+]i) in macula densa (MD) cells were measured with microelectrodes and microscope photometry, respectively. Our data suggest that the apical membrane of MD cell has a Ca2+ entry pathway, and that the basolateral membrane of MD cell has a Cl- conductance, which may be regulated by the [Ca2+]i in MD cell. We next evaluated by measuring the afferent arteriolar diameter with video camera monitoring whether the change in Ca2+ concentration of macula densa tubular lumen indeed affect tubuloglomerular feedback. The data suggest that the changes in luminal Ca2+ as well as Cl- concentration at macula densa independently alter the afferent arteriolar diameter, thus may affect the single nephron glomerular filtration rate.
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