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. 2022 Feb 14;9(1):ENEURO.0381-21.2021. doi: 10.1523/ENEURO.0381-21.2021

Figure 5.

Figure 5.

Chronic hM4Di-DREADD-mediated inhibition of CaMKIIα-positive forebrain excitatory neurons during the juvenile window does not influence anxiety, despair, or sensorimotor gating behavior in adulthood in CaMKIIα-tTA::TRE-hM4Di bigenic male mice. A, Shown is a schematic for the experimental paradigm for vehicle (5% sucrose) or CNO (5 mg/kg) administration in the juvenile window (P28 to P40) to CaMKIIα-tTA::TRE-hM4Di bigenic mice, which were then assessed for anxiety-like behaviors 2 months postcessation of CNO treatment in adulthood in the male cohort. B, Shown are representative tracks for vehicle-treated (top panel) and JCNO-treated CaMKIIα-tTA::TRE-hM4Di bigenic male mice (bottom panel) in the open field arena. C–F, No significant difference was noted between vehicle- and JCNO-treated male mice in the total distance traveled in the arena (C), the percentage of time spent in the center (D), the percentage of distance traveled in the center (E), or the total number of entries to the center of the open field arena (F; n = 18 for vehicle-treated male mice; n = 16 for JCNO-treated male mice). G, Shown are representative tracks for vehicle-treated (top panel) and JCNO-treated CaMKIIα-tTA::TRE-hM4Di bigenic male mice (bottom panel) in the elevated plus maze. H–M, No significant difference was observed between vehicle- and JCNO-treated male mice in the percentage of time spent in the closed (H) or open (I) arms of the EPM, as well as the percentage of distance traveled in closed (J) or open (K) arms, and for the number of entries to the closed (L) or open (M) arms (n = 18 for vehicle-treated male mice; n = 16 for JCNO-treated male mice). N, Shown are representative tracks for vehicle-treated (top panel) and JCNO-treated (bottom panel) CaMKIIα-tTA::TRE-hM4Di bigenic male mice in the LD box. O, P, No significant difference was noted between vehicle- and JCNO-treated male mice in either the total time spent in (O) or the total number of entries (P) into the light chamber of the LD box (n = 18 for vehicle-treated male mice; n = 16 for JCNO-treated male mice). Q, Shown is a schematic for the FST tank. R, S, No significant difference was noted between vehicle- and PNCNO-treated male mice for the percentage of immobility time (R) or for the total number of immobility events (S; n = 11 for vehicle-treated male mice; n = 11 for JCNO-treated male mice). T, Shown is a schematic for the protocol used for PPI to assess sensorimotor gating responses in adult male mice. PPI testing was conducted as described in Materials and Methods with basal startle determined following habituation, and PPI determined for +4 dB (69 dB), +8 dB (73 dB), and +16 dB (81 dB) above background noise (65 dB), followed by exposure to 120 dB for final startle. U, Basal startle response in JCNO-treated CaMKIIα-tTA::TRE-hM4Di bigenic male mice was unaltered compared with vehicle-treated CaMKIIα-tTA::TRE-hM4Di bigenic male controls. V, CaMKIIα-tTA::TRE-hM4Di bigenic male mice with a history of JCNO treatment did not show any change in PPI compared with the vehicle-treated controls (n = 10 for vehicle; n = 9 for JCNO male mice). Results are expressed as the mean ± SEM, and groups are compared using the two-tailed, unpaired Student’s t test.