Ectopic expression of mouse melanopsin in Drosophila photoreceptors reveals fast response kinetics and persistent dark excitation [Neurobiology]

The intrinsically photosensitive retinal ganglion cells (ipRGC) initiate non-image forming light-dependent activities and express the melanopsin (OPN4) photopigment. Several features of ipRGC photosensitivity are characteristic of fly photoreceptors. However, the light-response kinetics of ipRGC is much slower due to unknown reasons. Here we used transgenic Drosophila, in which the mouse OPN4 replaced the native Rh1 photopigment of Drosophila R1-6 photoreceptors, resulting in deformed rhabdomeric structure. Immunocytochemistry revealed OPN4 expression at the base of the rhabdomeres, mainly at the rhabdomeral stalk. Measurements of the Early Receptor Current (ERC), a linear manifestation of photopigment activation indicated large expression of OPN4 in the plasma membrane. Comparing the ERC amplitude and action spectra between wild type (WT) and the Opn4-expressing Drosophila, further indicated that large quantities of a blue absorbing photopigment were expressed, having a dark-stable blue intermediate state. Strikingly, the light induced current (LIC) of the Opn4-expressing fly photoreceptors was ~130 folds faster than that of ipRGC. Furthermore, intense white flash induced small amplitude prolonged dark current composed of discrete unitary currents similar to the Drosophila single photon responses. The induction of prolonged dark currents by intense blue light could be suppressed by a following intense green light, suggesting induction and suppression of Prolonged Depolarizing Afterpotential (PDA). This is the first demonstration of heterologous functional expression of mammalian Opn4 in the genetically emendable Drosophila photoreceptors. Moreover, the fast Opn4-induced ionic current relative to that of mouse ipRGC, indicates that the slow light-response of ipRGC does not arise from an intrinsic property of melanopsin.

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