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. Author manuscript; available in PMC: 2019 May 7.
Published in final edited form as: Nat Neurosci. 2018 Mar 5;21(4):638–646. doi: 10.1038/s41593-018-0091-7

Figure 4. Membrane properties of neurons transduced with second-generation rabies viral vectors remain normal for at least eight weeks.

Figure 4.

(a) Confocal images of basolateral amygdala neurons projecting to the nucleus accumbens retrogradely labeled with either retrobeads (RB, left panels), first generation rabies virus expressing Cre (ΔG, middle panels), or second generation of rabies virus expressing Cre (ΔGL, right panels), imaged after targeted whole-cell recording. (b) Representative traces of a seal test response in voltage-clamp, and (c) a ramp test to induce action potential firing in current-clamp mode. (d) The series resistance (Rs), capacitance (Cm), membrane resistance (Rm), decay time constant (Tau), and (e) holding current at −70 mV were not different between the three experimental groups (one-way ANOVA, F(2,36)=2.51, p=0.096 (Rs); F(2,36)=1.35, p=0.27 (Cm); F(2,36)=1.38, p=0.26 (Rm); F(2,36)=0.126, p=0.88 (Tau) – n=13(RB), 15(ΔG), and 11(ΔGL); one-way ANOVA, F(2,35)=3.14, p=0.056 (holding current) – n=13(RB), 14(ΔG), and 11(ΔGL)). (f) The action potential (AP) threshold was significantly more negative for the surviving cells transduced with the first generation RV compared to cells containing RB while cells transduced with the second generation RV did not show a difference compared to RB containing cells (one-way ANOVA, F(2,36)=4.29, *p=0.021, RB vs ΔG **p<0.05, RB vs ΔGL p>0.05; n=13(RB), 15(ΔG), and 11 (ΔGL)). The minimal current necessary to induce firing (rheobase) was not significantly different between the three groups (one-way ANOVA, F(2,36)=0.531, p=0.59; n=13(RB), 15(ΔG), and 11(ΔGL)). All bar graphs display mean ± s.e.m. Experiments in this figure were performed in 6 animals with similar results in all cases.