(a–c) whole-cell recording and immunofluorescence (IF) post hoc identification of ERα in recorded cells in OVX + E AVPV-AAV-Esr1 infected mice. (a) visualization during recording; (b) representative depolarizing (+20 pA, magenta) and hyperpolarizing (−20 pA, black) firing signatures. (c) neurobiotin (blue) and ERα (red) staining after photobleaching of GFP and mCherry signals. From top to bottom: cells not infected by AVPV-AAV-Esr1 and immunopositive for ERα; cells infected by AVPV-AAV-Esr1 but still immunopositive for ERα; cells infected by AAV-Esr1 and not immunopositive for ERα; cells infected by AVPV-AAV-LacZ and immunopositive for ERα. (d) left, input-output curves of infected cells with undetectable ERα in AAV-Esr1 (third row in a-c, black circle), cells infected by AVPV-AAV-lacZ (bottom row in a-c, white circle, n = 14), and cells not infected by AAV (top row in a-c, orange circle); middle, input-output curves from a separate set of cells in which Esr1 status was confirmed by single-cell qPCR post hoc (AAV-Esr1 black circle; AAV-lacZ, white circle); right, input-output curve of AVPV-AAV-Esr1 knockdown (black circle) vs KERKO (yellow circle) cells. (e,f) percent of cells exhibiting DIB (e) or rebound bursts (f). Cells per group is shown within or on top of the bar. (g), representative action potentials at the rheobase from lacZ vs Esr1 infected cells. (h–j) individual values and mean ± SEM rate of rise (h) full width at half maximum (FWHM) (i) and afterhyperpolarization potential amplitude (AHP) (j). *p<0.05 vs all other groups; # p<0.05 vs uninfected.