Biophysical Basis of the in vivo Electroretinogram of the Mouse: Current Source Density Analysis of Genetically and Pharmacologically Isolated Rod photoreceptor-driven Currents

To better understand the molecular basis of the mouse electroretinogram (ERG) we have developed a biophysical model of the rod photoreceptor layer’s ionic mechanisms and applied current source-density (CSD) analysis to predict the genetically and pharmacologically isolated rod ERG a -wave. The saturating a -wave is characterized by a rapid relaxation (τ ~ 6 ms) from its maximum that has been hypothesized to be caused by HCN1 channel opening consequent to light-triggered sustained hyperpolarization, or by extracellular flow of capacitive current during hyperpolarization. To test these hypotheses, the CSD model included an ensemble of 11 rods with cell body locations spanning the outer nuclear layer, and with ionic mechanisms -- including CNG channels, NCKX in the outer segment, NKX, Kv2.1 channels in the inner segment, and HCN1 throughout the “non-outer segment” -- fully specified as to axial distributions, voltage dependencies, and the extracellular conductivity of the photoreceptor layer extracellular space. Predicting with CSD the steady-state axial distributions of rod dark current and transretinal potential, and the spatio-temporal response of the activation phase of the rod’s light response to intense stimuli, the analysis confirmed the Robson-Frishman hypothesis that extracellular capacitive current flowing towards the inner segment upon the rapid closure of CNG current plays a major role in shaping the early a -wave. The CSD analysis also reveals that extracellular current from HCN1 channels opened by the hyperpolarization contributes a net extracellular current flowing toward the ONL that contributes materially to the saturated a -wave relaxation and to setting its plateau level.