Beta-adrenergic stimulation of Na(+)-K(+)-2Cl(-) cotransport activity in the rabbit lens.
Experimental maneuvers known to increase cellular cAMP levels evoked a stimulation in the K(+) influx across the anterior surfaces of isolated rabbit lenses, as measured by 86Rb(+) uptake. For this, the lenses were mounted in a modified Ussing-type chamber and exposed to the radiolabel under short-circuit conditions. The enhanced, cAMP-elicited flux was attributed to the basolateral Na(+)-K(+)-2Cl(-) cotransporter given its preclusion by bumetanide, a highly selective inhibitor of this symport, and the ineffectiveness of ouabain in mitigating the stimulation. The ouabain- plus bumetanide-insensitive K(+) uptake, which is about 10% of the total influx and represents passive entry of the radiolabel, was not affected by cAMP-elevating conditions. Forskolin, an activator of adenylyl cyclase; epinephrine, a non-selective adrenergic agonist; and the beta-selective agents, isoproterenol and terbutaline, were among the drugs used to elicit the increase in bumetanide-sensitive K(+) inflow. In experiments with isoproterenol, the stimulated influx evoked by the agonist was inhibited in lenses simultaneously exposed to propranolol. Other observations included that the stimulation of bumetanide-sensitive K(+) influx with forskolin was eliminated in lenses pretreated with the protein kinase inhibitors, staurosporine or H-89. However, these drugs were ineffective in preventing the increased influx produced by calyculin A, a phosphatase inhibitor, suggesting modulation of the cotransporter by at least two independent pathways. The cAMP-generating stimuli also produced an inhibition of the short-circuit current across the lens and an increase in translens resistance. These latter effects suggest that cAMP elevation also evokes an inhibition in an epithelial conductance(s) simultaneously to the stimulation of the cotransporter. As such, this study provides the first indication for the regulation of lens transport by adrenoceptors, presumably of the beta-2 subtype.
Alvarez
Lawrence J
LJ
Department of Ophthalmology, Mount Sinai School of Medicine, New York, NY 10029-6574, USA. larry.alvarez@mssm.edu
Candia
Oscar A
OA
Polikoff
Lee A
LA
D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D000818:Animals; D002034:Bumetanide / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D046508:Culture Techniques; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D007545:Isoproterenol / Q000494:pharmacology; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism*; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D011817:Rabbits; D028021:Sodium-Potassium-Chloride Symporters / Q000187:drug effects / Q000378:metabolism*
D000318:Adrenergic beta-Agonists; D028021:Sodium-Potassium-Chloride Symporters; D002034:Bumetanide; D005576:Colforsin; D010042:Ouabain; D000242:Cyclic AMP; D007545:Isoproterenol; D011188:Potassium