This indirect effect results from the fact that cortical Pyr cells within layer 2/3 are recurrently connected; thus, an increase in firing rate of Pyr cells in response to PV cell suppression (as observed above) may lead to an increase in the amount of excitation received by the Pyr cells themselves. To quantify the net Epacadostat clinical trial decrease in visually evoked inhibition during Arch-mediated suppression of PV cells we recorded in the whole-cell voltage-clamp configuration from layer 2/3 Pyr cells (targeted with two-photon microscopy) using a Cs-based internal solution. When the membrane potential of Pyr cells was clamped at the reversal potential of glutamate-mediated synaptic excitation

(∼15mV), photo suppression of PV cells decreased by 10% the postsynaptic inhibitory currents evoked by visual stimuli in Pyr cells (−9% ± 20%; n = 13 cells, p < 0.03; Figure 5A). To quantify the impact of PV cell suppression on excitation, Pyr cells were voltage clamped at the reversal potential for GABAA receptor-mediated inhibition (−80mV). Photo suppression of PV cells led to a small but significant increase in spontaneous excitatory conductance (0.1 ± 0.02 nS; n = 10; p < 0.004), demonstrating that our recordings are indeed sensitive to changes in excitation. However no significant increase was measured in

visually evoked excitatory GSK1349572 chemical structure conductance (n = 10; p = 0.5; Figure 5B). Thus, PV cell suppression results in little change in excitation but a net decrease in synaptic inhibition on to Pyr cells. Can this relatively small decrease in inhibition explain the observed linear transformation of Pyr cell spiking activity? To test this however we constructed a simple conductance-based model of Pyr cell spiking activity and studied its dependence on stimulus

orientation. To fully capture the linear transformation, not only must the decrease in inhibition result in a robust ∼ 40% increase in Pyr cells response, but it must do so while having only slight impact on tuning properties and, in particular, tuning sharpness. To set up the fundamentals of the model we first considered the orientation tuning under control conditions. To this end, we recorded excitatory and inhibitory conductances in layer 2/3 Pyr cells as a function of orientation. Stimulus-evoked excitatory currents (Figure 5C, red trace) recorded at the reversal potential for GABAA receptor-mediated inhibition showed clear tuning: they were on average 1.7-fold (n = 4) larger at the preferred orientation than at the nonpreferred orientation. In contrast, inhibitory currents (Figure 5C, blue trace) recorded at the reversal potential of glutamate-mediated synaptic excitation were less tuned, being only 1.2-fold (n = 5) larger at the preferred compared to the nonpreferred orientation (consistent with Liu et al., 2010).