The general procedure for animal preparation, anesthesia, ERG recording, light stimulation and data analysis has been previously described in detail in Della Santina et al. [19] (link). Briefly: ERGs were recorded in complete darkness via coiled gold electrodes making contact with the moist cornea. A small gold plate placed in the mouth served as both reference and ground. HCN inhibition was induced by subcutaneous injections of 12 mg/kg ivabradine. Responses were amplified differentially, band-pass filtered at 0.1 to 500 Hz, digitized at 12.8 kHz by a computer interface (LabVIEW 6.1; National Instruments, Austin, TX) and stored on disc for processing. Responses to flashes were averaged with an interstimulus interval ranging from 60 s for dim lights to 120 s for the brightest flashes.
The full field illumination of the eyes was achieved via a Ganzfeld sphere 30 cm in diameter, whose interior surface was coated with a highly reflective white paint. Two stimulus patterns were adopted: brief flashes that generated the typical ERG response (a- and b-waves) and sinusoidal time varying luminance stimuli eliciting periodic responses.
The full field illumination of the eyes was achieved via a Ganzfeld sphere 30 cm in diameter, whose interior surface was coated with a highly reflective white paint. Two stimulus patterns were adopted: brief flashes that generated the typical ERG response (a- and b-waves) and sinusoidal time varying luminance stimuli eliciting periodic responses.
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