Peak scaled NSFA to estimate single-channel conductance (g) and number of receptors (N). A, Variance (blue, right ordinate) was calculated by subtracting the scaled mean current (I_mean, red) from the current during an individual event (black). B, Top, Mean variance (difference squared, green). Bottom, Mean current (red). C, Estimation of N and g using a parabolic fit (purple) to the binned variance (black dots) plotted as a function of I_mean. Variance = i I_mean − I_mean²/N. Single-channel conductance, g, is calculated from the single-channel current (i) and the driving force. D, NSFA of arcuate nucleus kisspeptin mPSCs that responded with a change in amplitude in the presence of diazepam (n = 5 of 6). Data are mean ± SEM (N and g). *p < 0.05. E, NSFA of arcuate nucleus kisspeptin mPSCs reveals that estradiol dampens mPSC amplitude by decreasing the number of receptors bound by GABA. Data are mean ± SEM (N and g). *p < 0.05. n = 8 OVX PM, 6 OVX+E PM (one animal per recording). F, Data are mean ± SEM (N and g) in AVPV kisspeptin neurons. n = 5 OVX PM, 4 OVX+E PM (one animal per recording).