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Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias) III. Effects of stimulation of secretion by cyclic AMP

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Greger,  Rainer
Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society;

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Schlatter,  Eberhard
Department of Physiology, Max Planck Institute of Biophysics, Max Planck Society;

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Greger, R., Schlatter, E., Wang, F., & Forrest Jr., J. N. (1984). Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias) III. Effects of stimulation of secretion by cyclic AMP. Pflügers Archiv: European Journal of Physiology, 402, 376-384. doi:10.1007/BF00583938.


Cite as: http://hdl.handle.net/21.11116/0000-0008-1322-C
Abstract
Segments of rectal gland tubules (RGT) the spiny dogfish (Squalus acanthias) were perfused in vitro to study the cellular mechanism by which NaCl secretion is stimulated. Transepithelial PD (PDte), transepithelial resistance (Rte), the PD across the basolateral membrane (PDbl), the fractional resistance of the lumen membrane (FR1), and the cellular activities for Cl, Na+, and K+ (a cellx ) were measured. In series 1 the effects of stimulation (S) (dbcAMP 10−4, adenosine 10−4, and forskolin 10−6 mol · l−1) on these parameters were recorded and compared to nonstimulated state (NS). PDte increased from −1.9±0.2 mV to −11.0±0.9 mV (n=51). PDbI depolarized from −86±1 to −74±1.4 mV (n=52). Rte fell from 29±2.8 to 21±2 ωcm2 (n=23), and FR1 fell from 0.96±0.005 to 0.79±0.04 (n=9).a cellK+ was constant (123±13 versus 128±17 mmol · 1−1) (n=6), buta cellCl -fell significantly from 48±4 to 41±3 mmol · l−1 (n=7).a cellNa+ increased from 11±2.1 to 29.5±6.6 mmol · l−1 (n=4). In series 2 the conductivity properties were examined by rapid K+, and Cl concentration steps on the basolateral and luminal cell side respectively in NS and S states. In NS-segments reduction of bath K+ led to a hyperpolarization of PDbI with a mean slope of 28±1.3 mV/decade (n=9) (as compared to 19 mV/decade for S-state). Reduction of lumen Cl led to very little depolarization of the lumen membrane PD in NS-state: 6.5±2.3 mV/decade (n=4) (as compared to 13 mV/decade for S-state). In series 3 the effects of furosemide (7 · 10−5 mol l−1, bath) were examined in NS and S tubules. In NS RGT segments furosemide had no effect on PDbI or PDte;a cellCl fell slowly after furosemide with an initial rate of 0.33 mmol · l1 s−1, as compared to 1.5 mmol · l−1 · s−1 for S-state. The increase ina cellCl after removal of furosemide from NS to S-states was examined in the presence of furosemide. Despite the presence of furosemide stimulation was accompanied by a fall in Rte, FR1, anda cellCl . From these data we conclude that (a) stimulation by cyclic AMP increases the Cl-conductance of the apical cell membrane at least by a factor of 10, that (b) in the NS-state the Na+2ClK+ carrier can be triggered to work at rates similar to the S state by lowering a cellCl , and that (c) the increase in apical Cl-conductance is the primary event in cyclic AMP mediated stimulation of NaCl secretion.