Regulation of the Na+2ClK+ cotransporter in in vitro perfused rectal gland tubules of Squalus acanthias.
Pflugers Arch
; 436(4): 521-8, 1998 Jul.
Article
em En
| MEDLINE
| ID: mdl-9683724
ABSTRACT
Previously it has been shown that the Na+2ClK+ cotransporter accepts NH4 + at its K+ binding site. This property can be used to estimate its transport rates by adding NH4 + to the bath and measuring the initial furosemide-dependent rates of change in BCECF fluorescence. We have utilized this technique to determine the regulation of the furosemide-inhibitable Na+2ClK+ cotransporter in in vitroperfused rectal gland tubules (RGT) of Squalus acanthias. Addition of NH4 + to the bath (20 mmol/l) led to an initial alkalinization, corresponding to NH3 uptake. This was followed by an acidification, corresponding to NH4 + uptake. The rate of this uptake was quantified by exponential curve fitting and is given in arbitrary units (Δfluorescence/time). This acidification could be completely inhibited by furosemide. In the absence of any secretagogue preincubation of RGT in a low Cl solution (6 mmol/l, low Cl) for 10 min enhanced the uptake rate significantly from 4.04±0.51 to 12.7±1.30 (n=5). The addition of urea (200 mmol/l) was without effect, but the addition of 300 mmol/l mannitol (+300 mannitol) enhanced the rate significantly from 7.24±1.33 to 14.7±4.6 (n=6). Stimulation of NaCl secretion by a solution maximizing the cytosolic cAMP concentration (Stim) led to a significant increase in NH4 + uptake rate from 5.00±1.33 to 13.3±1.54 (n=6). Similar results were obtained in the additional presence of Ba2+ (1 mmol/l) the uptake rate was increased significantly from 4.23±0.34 to 15.1±1.86 (n=16). In the presence of Stim low Cl had no additional effect on the uptake rate 15.1±3.1 versus 15.2±2.8 in high Cl (n=6). The uptake rate in Stim containing additional +300 mannitol (22.3±4.0, n=5) was not significantly different from that obtained with Stim or +300 mannitol alone. By whatever mechanism the NH4 + uptake rate was increased furosemide (500 µmol/l) always reduced this rate to control values. Hence three manoeuvres enhanced furosemide-inhibitable uptake rates of the Na+2ClK+ cotransporter probably independently (1) lowering of cytosolic Cl concentration; (2) cell shrinkage; and (3) activation by cAMP.
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Base de dados:
MEDLINE
Assunto principal:
Glândula de Sal
/
Simportadores de Cloreto de Sódio-Potássio
/
Squalus acanthias
Idioma:
En
Ano de publicação:
1998
Tipo de documento:
Article