Functional modulation of phosphodiesterase-6 by calcium in mouse rod photoreceptors

Phosphodiesterase-6 (PDE6) is a key protein in the G-protein cascade converting photon information to bioelectrical signals in vertebrate photoreceptor cells. Here, we demonstrate that PDE6 is regulated by calcium, contrary to the common view that PDE1 is the unique PDE class whose activity is modulated by intracellular Ca²⁺. To broaden the operating range of photoreceptors, mammalian rod photoresponse recovery is accelerated mainly by two calcium sensor proteins: recoverin, modulating the lifetime of activated rhodopsin, and guanylate cyclase-activating proteins (GCAPs), regulating the cGMP synthesis. We found that decreasing rod intracellular Ca²⁺ concentration accelerates the flash response recovery and increases the basal PDE6 activity (β_dark) maximally by ~ 30% when recording local electroretinography across the rod outer segment layer from GCAPs^-/- recoverin^-/- mice. Our modeling shows that a similar elevation in β_dark can fully explain the observed acceleration of flash response recovery in low Ca²⁺. Additionally, a reduction of the free Ca²⁺ in GCAPs^-/- recoverin^-/- rods shifted the inhibition constants of competitive PDE inhibitor 3-isobutyl-1-methylxanthine (IBMX) against the thermally activated and light-activated forms of PDE6 to opposite directions, indicating a complex interaction between IBMX, PDE6, and calcium. The discovered regulation of PDE6 is a previously unknown mechanism in the Ca²⁺-mediated modulation of rod light sensitivity.