The sequence (a) priming flash, (b) dark interval, and (c) red light induces a long-lasting afterdepolarization (PDA) in Balanus photoreceptors. The inward flow of membrane current associated with the decay of PDA was independent of red test flashes, provided that PDA had plateaued at a particular intensity. The influence of wavelength and duration of the priming flash and their interaction with the dark interval were investigated. Increasing the duration of the priming flash produced a systematic increase in PDA duration. The dark interval plays a crucial role in PDA induction. The priming flash duration and the dark interval were reciprocally related, i.e, short flashes followed by long dark intervals induced as much PDA as long priming flashes followed by short dark intervals. The action spectrum for the priming flash was found to correspond to that of the primary photopigment (VP537).

Anion and cation permeabilities in dark-adapted Balanus photoreceptors were determined by comparing changes in the membrane potential in response to replacement of the dominant anion (Cl-) or cation (Na+) by test anions or cations in the superfusing solution. The anion permeability sequence obtained was PI greater than PSO4 greater than PBr greater than PCl greater than Pisethionate greater than Pmethanesulfonate. Gluconate, glucuronate, and glutamate generally appeared more permeable and propionate less permeable than Cl-. The alkali-metal cation permeability sequence obtained was PK greater than PRb greater than PCx greater than PNa approximately PLi. This corresponds to Eisenman's IV which is the same sequencethat has been obtained for other classes of nerve cells in the resting state. The values obtained for the permeability ratios of the alkali-metal cations are considered to be minimal. The membrane conductance measured by passing inward current pulses in the different test cations followed the sequence, GK greater than GRb greater than GCs greater than GNa greater than GLi. The conductance ratios obtained for a full substitution of the test cation agreed quite well with permeability ratios for all the alkali-metal cations except K+ which was generally higher.

When intracellular chloride activity (aiCl) was monitored with chloride-sensitive liquid ion exchanges (CLIX) microelectrodes in Balanus photoreceptors, replacement of extracellular chloride (Cl0) by methanesulfonate or glutamate was followed by a rapid but incomplete loss of aiCl. When propionate was used as the extracellular anion substitute, CLIX electrodes detected an apparent gain in aiCl, while a newly designed Ag-AgCl wire-in glass microelectrode showed a loss of aiCl under the same conditions. This discrepancy in Cl- washout when propionate replaced Cl0 is explained by the differences in selectivity of CLIX and Ag-AgCl electrodes for native intracellular anions and for the extracellular anion substitute which also replaces Cli and interferes in the determination of aiCl. Both electrodes indicate that ECl approximately Em when the cells are bathed in normal barnacle saline, and both electrodes showed the rate of Cl washout (tau approximately 5 min) to be independent of Cli when Cl0 was replaced by glutamate. Details of Ag-AgCl microelectrode construction are presented. These electrodes were tested and found to be insensitive to the organic anion substitutes used in this study. Selectivity data of CLIX electrodes for several anions of biological interest are described.

Ion-sensitive microelectrodes were used to measure intracellular activities (aix) of Na+, K+, and C-1 in Balanus photoreceptors. Average values of aiNa, aiK, and aiCl were 28 mM, 120 mM, and 65 mM, respectively. Equilibrium potentials calculated from these average values were: Na+ +64 mV, K+ - 77 mV, and and Cl- -42 mV; ther average value of the resting potential for all cells examined was -41 mV. Long exposure to intense illumination produced measurable increases in aiNa. Classical Na+ - K+ reciprocal dilution experiments were analyzed with and without observed changes in aiK. As aoK was increased, the membrane depolarized, and aiK increased. Better agreement was found between the membrane potential and the directly determined EK than expected from the standard relation between Em and aoK. The latter produced pNa:pK estimates of the resting photoreceptor membrane that were higher than estimates based on data from the ion electrodes. Generally, Em was more negative than EK as aoK was increased. This is consistent with a significant chloride permeability in the dark-adapted photoreceptor.