Changes in the light-sensitive current of salamander rods upon manipulation of putative pH-regulating mechanisms in the inner and outer segment.

ABSTRACT

The light-sensitive current of dark-adapted rods isolated from the Ambystoma retina was recorded while either the inner or the outer segment (IS or OS) protruding from the suction pipette was exposed to treatments intended to reveal the physiological roles of pH-regulating transport mechanisms. Applied to the IS, both amiloride (presumed to block Na+/H+ exchange, 2 mM) and 4-4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) (presumed to block bicarbonate transport, 0.1 mM) generally abolished light sensitivity completely but reversibly, consistent with acidification of the IS. Yet, the circulating ("dark") current often persisted, implying that the OS was not acidified. Applied to the OS, amiloride depressed but DIDS increased the dark current and photoresponses. Given the fact that the current increases with rising OS-pHi, this suggests alkalinization, which could be due to DIDS inhibiting bicarbonate extrusion by HCO3-/Cl- exchangers in the OS. Consistent with this idea, replacing external Cl- by other anions increased the current as would be expected if HCO3-/Cl- exchange is reversed. We propose that the IS and OS manage their acid balances independently and with different sets of transport mechanisms. Acidosis in either compartment suppresses the photosensitivity of the rod, but by differing mechanisms.