The rapidly acting antidepressants ketamine and scopolamine exert behavioral effects that can last from several days to more than a week in some patients. The molecular mechanisms underlying the maintenance of these antidepressant effects are unknown. Here we show that methyl-CpG-binding protein 2 (MeCP2) phosphorylation at Ser421 (pMeCP2) is essential for the sustained, but not the rapid, antidepressant effects of ketamine and scopolamine in mice. Our results reveal that pMeCP2 is downstream of BDNF, a critical factor in ketamine and scopolamine antidepressant action. In addition, we show that pMeCP2 is required for the long-term regulation of synaptic strength after ketamine or scopolamine administration. These results demonstrate that pMeCP2 and associated synaptic plasticity are essential determinants of sustained antidepressant effects.
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We thank members of the Monteggia and Kavalali laboratory for helpful advice and discussions on the manuscript. We thank K. Szabla for the initial insight into this project. We thank A. West for providing the initial pMeCP2 antibody for these studies. This work was supported by National Institutes of Health grants MH070727 and MH081060 (to L.M.M.) and MH066198 (to E.T.K.); the Basic Science Research Program through the National Research Foundation of Korea, funded by the Ministry of Education (2016R1A6A3A03008533, to J.K.); and postdoctoral fellowships from the Elisabeth and Alfred Ahlqvist Foundation within the Swedish Pharmaceutical Society and the Swedish Society for Medical Research (to C.B.).
C.B. is currently employed by Janssen-Cilag AB, Sweden, but contributed to this work in his previous position at UTSW. Janssen-Cilag AB had no part in the planning, execution or funding of this study. The other authors declare no competing financial interests.
Peer review information Nature Neuroscience thanks the anonymous reviewers for their contribution to the peer review of this work.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended Data Fig. 1 pMeCP2 is not changed in the medial prefrontal cortex 7 days after ketamine treatment.
pMeCP2 levels were measured with Western blot analysis in the medial prefrontal cortex of C57BL/6J mice 7 days after ketamine treatment. No significant changes in pMeCP2 levels were observed between saline- and ketamine-treated groups. (two-sided unpaired t-test, ketamine: t(14) = 0.3298, P = 0.7464, n = 8 mice per group). In Western blot analysis for pMeCP2, membrane above about 70 KDa was cropped and used for immunoblotting. The graph represents mean ± S.E.M., N.S.: not significant, Sal: saline, Ket: ketamine. For detailed statistical information, see Supplementary Table 1.
Extended Data Fig. 2 Ketamine rapidly increases BDNF protein levels in the hippocampus of Mecp2 S421A KI mice.
BDNF protein levels were measured 30 min after ketamine treatment in Mecp2 KI and CTL mice by Western blot analysis. BDNF levels were significantly increased by ketamine treatment in both the CTL and KI mice (two-way ANOVA with Tukey’s multiple comparisons, Genotype x Drug: F(1, 21) = 0.0157, P = 0.9016, Genotype: F(1, 21) = 0.0115, P = 0.9158, Drug: F(1, 21) = 31.93, P < 0.0001, CTL-Sal, CTL-Ket, KI-Sal, KI-Ket: n = 7, 7, 6, 5 mice). In Western blot analysis for pMeCP2, membrane above about 70 KDa was cropped and used for immunoblotting. The graph represents mean ± S.E.M., **, P < 0.01, Sal: saline, Ket: ketamine, CTL: littermate control. For detailed statistical information, see Supplementary Table 1.
Extended Data Fig. 3 Effects of ketamine on pre-and post-synaptic function at 7 days after treatment.
a, Saline or ketamine was administered to C57BL/6J mice. The mice were sacrificed 7 days later, and slices were prepared for recordings. I-O curves were measured during baseline recording in the SC-CA1 synapses. I-O curves of Sal and Ket groups were from baseline recording in the Sal-Ket group of Fig. 3b and Ket-Ket group of Fig. 3c, respectively. The slope of I-O curves in the previous ketamine-treated group was not significantly different compared to the saline-treated group (two-sided unpaired t-test, t(17) = 1.448, P = 0.1659, Sal, Ket: n = 9, 10 slices). b, PPRs were measured before and after ketamine perfusion onto hippocampal slices of mice given either saline or ketamine 7 days before the slice preparation. PPRs were not significantly changed by either previous ketamine injection or subsequent ketamine perfusion (two-way ANOVA with Tukey’s multiple comparisons, all P-values > 0.05, Sal-Ket, Ket-Ket: n = 11, 10 slices). Representative traces are from data recorded at 30 msec interstimulus interval in the respective treatment groups. Graphs represent mean ± S.E.M., N.S.: not significant, Sal: saline, Ket: ketamine. Sal+Ket: ketamine perfusion onto hippocampal slices from saline-injected mice, Ket+Ket: ketamine perfusion onto hippocampal slices from ketamine-injected mice. For detailed statistical information, see Supplementary Table 1.
a,b, Ketamine-potentiation was measured in the Schaffer collateral to CA1 synapses of C57BL/6J female mice given either saline (a) or ketamine (b) injection 7 days before slice preparation. Augmented potentiation was observed in the group previously treated with ketamine (b: Ket+Ket, 138.5 ± 6.717%), compared to the group previously treated with saline (a: Sal+Ket, 114 ± 3.158%), (two-sided unpaired t-test, a - Sal+ACSF vs Sal+Ket: t(11) = 4.207, P = 0.0015, Sal+ACSF, Sal+Ket: n = 6, 7 slices, b - Ket+ACSF vs Ket+Ket: t(10) = 5.528, P = 0.0003, Ket+ACSF, Ket+Ket: n = 6, 6 slices, Sal+Ket in a vs Ket+Ket in b: t(11) = 3.465, P = 0.0053). Graphs represent mean ± S.E.M., **, P < 0.01, ***, P < 0.001, Sal: saline, Ket: ketamine. Sal+ACSF: ACSF perfusion onto hippocampal slices from saline-injected mice, Sal+Ket: ketamine perfusion onto hippocampal slices from saline-injected mice, Ket+ACSF: ACSF perfusion onto hippocampal slices from ketamine-injected mice, Ket+Ket: ketamine perfusion onto hippocampal slices from ketamine-injected mice. For detailed statistical information, see Supplementary Table 1.
Extended Data Fig. 5 Molecular changes in the CA1 hippocampal region at 3 or 7 days after ketamine treatment.
a,b, The hippocampal CA1 area was collected in C57BL6/J mice 3 or 7 days after ketamine treatment. Phosphorylation of CaMKIIβ, but not CaMKIIα, was significantly increased by ketamine at 3 days compared to the saline group, and the increased phosphorylation returned to the levels of the saline-treated group at 7 days (two-sided unpaired t-test, 3 days, CaMKIIα: t(14) = 0.4369, P = 0.6688, CaMKIIβ: t(14) = 3.756, P = 0.0021, n = 8 mice per group, 7 days, CaMKIIα: t(14) = 1.052, P = 0.3107, CaMKIIβ: t(14) = 0.3796, P = 0.7100, n = 8 mice per group). pMeCP2 levels were significantly increased at both 3 and 7 days (two-sided unpaired t-test, 3 days: t(16) = 2.332, P = 0.0331, n = 9 mice per group, 7 days: t(14) = 2.440, P = 0.0286, n = 8 mice per group). BDNF levels were not changed at 3 days (two-sided unpaired t-test, t(14) = 0.0433, P = 0.9661, n = 8 mice per group). In Western blot analysis for pMeCP2, membrane above about 70 KDa was cropped and used for immunoblotting. Graphs represent mean ± S.E.M., N.S.: not significant, *, P < 0.05, **, P < 0.01, Sal: saline, Ket: ketamine. For detailed statistical information, see Supplementary Table 1.
a, Immobility was measured in the FST 7 days after scopolamine treatment. The scopolamine-treated group showed a significant reduction in time spent immobile compared to the saline-treated group (two-sided unpaired t-test, t(17) = 2.469, P = 0.0244, Sal, SCA: n = 9, 10 mice). b, PPRs were analyzed in the hippocampal CA1 area of C57BL/6J mice given scopolamine 7 days prior. The PPRs were significantly reduced in the scopolamine-treated group compared to the saline-treated group at 20 and 30 msec interstimulus interval condition (two-sided Mann-Whitney test, 20 msec: U = 210, P = 0.0314, 30 msec: U = 207, P = 0.0270, Sal, SCA: n = 24, 27 slices). Graphs represent mean ± S.E.M., N.S.: not significant, *, P < 0.05, Sal: saline, SCA: scopolamine. For detailed statistical information, see Supplementary Table 1.
Actinomycin D was administered to C57BL/6J mice prior to the saline or scopolamine treatment, and the mice were tested 24 hrs later in the FST. Scopolamine did not significantly reduce the duration of immobility of the mice pretreated with actinomycin D (two-sided unpaired t-test, t(17) = 1.006, P = 0.3286, ActD-Sal, ActD-SCA : n = 10, 9 mice). The Graph represents mean ± S.E.M., N.S.: not significant, Sal: saline, SCA: scopolamine. For detailed statistical information, see Supplementary Table 1.
Extended Data Fig. 8 pMeCP2 is not changed in the medial prefrontal cortex 24 hrs after scopolamine treatment.
pMeCP2 levels were measured in the medial prefrontal cortex of C57BL/6J mice 24 hrs after scopolamine treatment with Western blot analysis. No significant differences in pMeCP2 levels were observed between saline- and scopolamine-treated groups (two-sided unpaired t-test, t(14) = 0.0705, P = 0.9448, n = 8 mice per group). In Western blot analysis for pMeCP2, membrane above about 70 KDa was cropped and used for immunoblotting. The Graph represents mean ± S.E.M., N.S.: not significant, Sal: saline, SCA: scopolamine. For detailed statistical information, see Supplementary Table 1.
pCREB levels were measured in the hippocampus of C57BL/6J mice with Western blot analysis 8 hrs and 24 hrs after scopolamine treatment. No significant differences were observed between Sal- and SCA-treated groups at either time point (two-sided unpaired t-test or Welch’s correct t-test, 8 hrs: t(9.272) = 0.9004, P = 0.3907, n = 8 mice per group, 24 hrs: t(12) = 0.0677, P = 0.9472, n = 7 mice per group). The Graph represents mean ± S.E.M., N.S.: not significant, Sal: Saline, SCA: Scopolamine. For detailed statistical information, see Supplementary Table 1.
Extended Data Fig. 10 Scopolamine increases Bdnf mRNA levels in the hippocampus of Mecp2 S421A KI mice 8 hours after injection.
Bdnf mRNA levels were measured with quantitative real-time PCR in CTL and Mecp2 KI mice 8 hrs after scopolamine. Scopolamine treatment significantly increased Bdnf mRNA levels in the hippocampus of both the CTL and KI mice (two-way ANOVA with Tukey’s multiple comparisons, Genotype x Drug: F(1, 33) = 0.0467, P = 0.8302, Genotype: F(1, 33) = 0.3223, P = 0.5741, Drug: F(1, 33) = 19.22, P = 0.0001, CTL-Sal, CTL-SCA, KI-Sal, KI-SCA: n = 10, 9, 10, 8 mice). The Graph represents mean ± S.E.M., *, P < 0.05, Sal: saline, SCA: scopolamine, CTL: littermate control. For detailed statistical information, see Supplementary Table 1.
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Kim, JW., Autry, A.E., Na, E.S. et al. Sustained effects of rapidly acting antidepressants require BDNF-dependent MeCP2 phosphorylation. Nat Neurosci 24, 1100–1109 (2021). https://doi.org/10.1038/s41593-021-00868-8
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