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. Author manuscript; available in PMC: 2024 Nov 27.
Published in final edited form as: Sci Signal. 2024 Oct 1;17(856):eado4132. doi: 10.1126/scisignal.ado4132

Figure 4. oAβ and GIRK channel inhibitor effect on excitatory synapses in cultured HPC neurons.

Figure 4.

(A and B) Cultured HPC neurons from male mice were transfected with the plasmids pAAV-hSyn-mCherry (to fill and label neuronal structures) and pAAV-Syn-PSD95.FingR-eGFP-CCR5TC (to label excitatory post-synaptic densities). Images were taken before (t=0) and after (24 and 48 hours) addition of oAβ (1 μM; A) or the GIRK channel inhibitor tertiapin (1 μM; B) to the media. Scale bar: 30 μm. Boxed regions in upper panels are displayed at higher magnification in lower panels. (C) Percent change in excitatory synapse count 24 and 48 hours in male HPC neurons treated with oAβ (A) or vehicle (V), or tertiapin (T) or vehicle (V). oAβ data were extracted from 21 (24 hours) or 17 (48 hours) ROIs and corresponding vehicle data were extracted from 17 (24 hours) or 14 (48 hours) ROIs. Tertiapin data were extracted from 17 ROIs (24 and 48 hours) and corresponding vehicle data were extracted from 23 (24 hours) or 19 (48 hours) ROIs. Data were derived from 8 coverslips per group pooled from 4 independent cultures. (D to F) As in (A to C), in HPC neurons from female mice. oAβ n = 24 (24 hours) or 18 (48 hours) ROIs and corresponding vehicle n = 19 (24 hours) or 18 (48 hours) ROIs; tertiapin n = 16 (24 hours) or 10 (48 hours) ROIs, and corresponding vehicle n = 23 (24 hours) or 19 (48 hours) ROIs. Data were derived from 8 coverslips per group pooled from 4 independent cultures. (G and H) sEPSCs in HPC neurons from male mice, measured 24 and 48 hours after treatment with oAβ (1 μM) or vehicle (G), or the GIRK channel inhibitor tertiapin (1 μM) or vehicle (H); scale: 100 pA/5s. (I and J) sEPSC amplitude (I) and frequency (J) summary in male HPC neurons from 10 (V/24 hours), 6 (A/24 hours), 10 (V/48 hours), and 6 (A/48 hours) neurons pooled from 3 independent cultures, as well as 7 (V/24 hours), 6 (T/24 hours), 7 (V/48 hours), 6 (T/48 hours) neurons pooled from 3 independent cultures. (K to N) As in (G to J) in HPC neurons from female mice. n = 7 (V/24 hours), 5 (A/24 hours), 8 (V/48 hours), and 6 (A/48 hours) neurons pooled from 3 independent cultures, as well as 6 neurons per group (V/T/24 hours/48 hours) pooled from 3 independent cultures. Data are presented in box-and-whisker format (min-max) (C,F) or mean ± SEM (I,J,M,N). Data were analyzed by two-way ANOVA with (C,F) or without (I,J,M,N) repeated measures (mixed-effects) and Šídák’s multiple comparisons, *P<0.05,** P<0.01,***P<0.001.