Functional interchangeability of rod and cone transducin α-subunits

Rod and cone photoreceptors use similar but distinct sets of phototransduction proteins to achieve different functional properties, suitable for their role as dim and bright light receptors, respectively. For example, rod and cone visual pigments couple to distinct variants of the heterotrimeric G protein transducin. However, the role of the structural differences between rod and cone transducin α subunits (Tα) in determining the functional differences between rods and cones is unknown. To address this question, we studied the translocation and signaling properties of rod Tα expressed in cones and cone Tα expressed in rods in three mouse strains: rod Tα knockout, cone Tα GNAT2cpfl³ mutant, and rod and cone Tα double mutant rd17 mouse. Surprisingly, although the rod/cone Tα are only 79% identical, exogenously expressed rod or cone Tα localized and translocated identically to endogenous Tα in each photoreceptor type. Moreover, exogenously expressed rod or cone Tα rescued electroretinogram responses (ERGs) in mice lacking functional cone or rod Tα, respectively. Ex vivo transretinal ERG and single-cell recordings from rd17 retinas treated with rod or cone Tα showed comparable rod sensitivity and response kinetics. These results demonstrate that cone Tα forms a functional heterotrimeric G protein complex in rods and that rod and cone Tα couple equally well to the rod phototransduction cascade. Thus, rod and cone transducin α-subunits are functionally interchangeable and their signaling properties do not contribute to the intrinsic light sensitivity differences between rods and cones. Additionally, the technology used here could be adapted for any such homologue swap desired.