1. Retinas from channel catfish were dissociated and the cells maintained in culture. Horizontal cells that normally receive input from cone photoreceptors were identified. The conductance of the electrical junction formed between a pair of ‘cone’ horizontal cells was measured by controlling the membrane voltage of each cell with a voltage clamp maintained through either a micropipette or a patch pipette. The two techniques yielded similar results. 2. Transjunctional current was measured while transjunctional voltage was stepped to values between +/‐ 60 mV. The current (measured 5 ms after a step) was proportional to voltage over the range tested. For steps to voltages greater than +/‐ 45 mV, the current exhibited a slight time‐dependent decline. 3. Dopamine decreased junctional conductance in a dose‐dependent fashion. A 50% reduction was obtained with 10 nM‐dopamine. The D1 agonist fenoldopam (100 nM) also decreased junctional conductance. The uncoupling produced by either agent was rapid and reversible. 4. The introduction of 100 microM‐cyclic AMP into one cell of a pair decreased junctional conductance by, on average, 40%. Forskolin (1‐10 microM), an activator of adenylate cyclase, decreased junctional conductance 50‐90%. 5. The introduction of 80 microM‐cyclic GMP into one cell of a pair decreased junctional conductance by, on average, 40%. Nitroprusside (1‐10 microM), an activator of guanylate cyclase, reduced junctional conductance 40‐65%. 6. The introduction of a peptide inhibitor specific for the cyclic AMP‐dependent protein kinase reversed a decrease in junctional conductance produced by superfusion with either dopamine (1 microM), fenoldopam (100 nM) or forskolin (5‐10 microM). 7. Intracellular Ca2+ concentration was measured with the fluorescent indicator Fura‐2. The intracellular Ca2+ concentration was increased by activation of a Ca2+ current. Junctional conductance remained constant as the internal Ca2+ concentration changed from 100 to 700 nM. 8. Intracellular pH was measured with the fluorescent indicator bis‐carboxyethylcarboxyfluorescein. The application of acetate (2.5 mM) reduced intracellular pH by 0.2‐0.3 units and decreased junctional conductance by approximately 50%. A subsequent application of fenoldopam did not alter intracellular pH, but decreased junctional conductance by more than 50%. 9. The sensitivity of the junctional conductance between isolated horizontal cells to dopamine is consistent with dopamine having a direct effect on coupling in intact retina. Dopamine regulates the activity of a cyclic AMP‐dependent protein kinase which in turn modulates junctional conductance. Changes in intracellular pH and Ca2+ concentration are not involved in mediating the effect of dopamine on coupling. Cyclic GMP and intracellular pH may participate in regulatory pathways independent of that used by cyclic AMP.
The Journal of Physiology – Wiley
Published: Jul 1, 1989
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