Ketamine was thought to induce rapid antidepressant responses by inhibiting GluN2B-containing N-methyl-d-aspartic acid (NMDA) receptors (NMDARs), which presents a promising opportunity to develop better antidepressants. However, adverse side effects limit the broader application of ketamine and GluN2B inhibitors are yet to be approved for clinical use. It is unclear whether ketamine acts solely through GluN2B-dependent mechanisms. The present study reports that the loss of another major NMDAR subunit, GluN2A, in adult mouse brains elicits robust antidepressant-like responses with limited impact on the behaviors that mimic the psychomimetic effects of ketamine. The antidepressant-like behavioral effects of broad NMDAR channel blockers, such as ketamine and MK-801 (dizocilpine), were mediated by the suppression of GluN2A, but not by the inhibition of GluN2B. Moreover, treatment with ketamine or MK-801 rapidly increased the intrinsic excitability of hippocampal principal neurons through GluN2A, but not GluN2B. Together, these findings indicate that GluN2A mediates ketamine-triggered rapid antidepressant-like responses.
This is a preview of subscription content, access via your institution
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
The datasets generated during and analyzed during the present study are available from the corresponding author on reasonable request. Source data are provided with this paper.
The present study did not use any customized code or mathematical algorithm.
Nierenberg, A. A. et al. Timing of onset of antidepressant response with fluoxetine treatment. Am. J. Psychiatry 157, 1423–1428 (2000).
Al-Harbi, K. S. Treatment-resistant depression: therapeutic trends, challenges, and future directions. Patient Prefer. Adherence 6, 369–388 (2012).
Duman, R. S. Ketamine and rapid-acting antidepressants: a new era in the battle against depression and suicide. F1000Res https://doi.org/10.12688/f1000research.14344.1 (2018).
Gibbons, R. D. et al. Early evidence on the effects of regulators’ suicidality warnings on SSRI prescriptions and suicide in children and adolescents. Am. J. Psychiatry 164, 1356–1363 (2007).
Kim, J., Farchione, T., Potter, A., Chen, Q. & Temple, R. Esketamine for treatment-resistant depression−first FDA-approved antidepressant in a new class. N. Engl. J. Med. 381, 1–4 (2019).
Ng, S. H., Tse, M. L., Ng, H. W. & Lau, F. L. Emergency department presentation of ketamine abusers in Hong Kong: a review of 233 cases. Hong Kong Med J. 16, 6–11 (2010).
Trullas, R. & Skolnick, P. Functional antagonists at the NMDA receptor complex exhibit antidepressant actions. Eur. J. Pharmacol. 185, 1–10 (1990).
Iosifescu, D. V. et al. Efficacy and safety of AXS-05 (dextromethorphan-bupropion) in patients with major depressive disorder: a phase 3 randomized clinical trial (GEMINI). J. Clin. Psychiatry https://doi.org/10.4088/JCP.21m14345 (2022).
Nowak, G., Ordway, G. A. & Paul, I. A. Alterations in the N-methyl-d-aspartate (NMDA) receptor complex in the frontal cortex of suicide victims. Brain Res. 675, 157–164 (1995).
Liu, R. et al. Correlation of functional GRIN2A gene promoter polymorphisms with schizophrenia and serum d-serine levels. Gene 568, 25–30 (2015).
Paoletti, P., Bellone, C. & Zhou, Q. NMDA receptor subunit diversity: impact on receptor properties, synaptic plasticity and disease. Nat. Rev. Neurosci. 14, 383–400 (2013).
Miller, O. H. et al. GluN2B-containing NMDA receptors regulate depression-like behavior and are critical for the rapid antidepressant actions of ketamine. eLife 3, e03581 (2014).
Jimenez-Sanchez, L., Campa, L., Auberson, Y. P. & Adell, A. The role of GluN2A and GluN2B subunits on the effects of NMDA receptor antagonists in modeling schizophrenia and treating refractory depression. Neuropsychopharmacology 39, 2673–2680 (2014).
Gerhard, D. M. et al. GABA interneurons are the cellular trigger for ketamine’s rapid antidepressant actions. J. Clin. Invest. 130, 1336–1349 (2020).
Henter, I. D., Park, L. T. & Zarate, C. A. Jr. Novel glutamatergic modulators for the treatment of mood disorders: current status. CNS Drugs 35, 527–543 (2021).
Zanos, P. et al. Ketamine and ketamine metabolite pharmacology: insights into therapeutic mechanisms. Pharm. Rev. 70, 621–660 (2018).
Aguilar-Valles, A. et al. Antidepressant actions of ketamine engage cell-specific translation via eIF4E. Nature 590, 315–319 (2021).
Cui, Y. et al. Astroglial Kir4.1 in the lateral habenula drives neuronal bursts in depression. Nature 554, 323–327 (2018).
Autry, A. E. et al. NMDA receptor blockade at rest triggers rapid behavioural antidepressant responses. Nature 475, 91–95 (2011).
Li, N. et al. mTOR-dependent synapse formation underlies the rapid antidepressant effects of NMDA antagonists. Science 329, 959–964 (2010).
Zanos, P. et al. NMDAR inhibition-independent antidepressant actions of ketamine metabolites. Nature 533, 481–486 (2016).
Wilkinson, S. T. & Sanacora, G. A new generation of antidepressants: an update on the pharmaceutical pipeline for novel and rapid-acting therapeutics in mood disorders based on glutamate/GABA neurotransmitter systems. Drug Discov. Today 24, 606–615 (2019).
Berberich, S. et al. Lack of NMDA receptor subtype selectivity for hippocampal long-term potentiation. J. Neurosci. 25, 6907–6910 (2005).
Francija, E. et al. GluN2A-ERK-mTOR pathway confers a vulnerability to LPS-induced depressive-like behaviour. Behav. Brain Res. 417, 113625 (2022).
Boyce-Rustay, J. M. & Holmes, A. Genetic inactivation of the NMDA receptor NR2A subunit has anxiolytic- and antidepressant-like effects in mice. Neuropsychopharmacology 31, 2405–2414 (2006).
Lim, A. L., Taylor, D. A. & Malone, D. T. Consequences of early life MK-801 administration: long-term behavioural effects and relevance to schizophrenia research. Behav. Brain Res. 227, 276–286 (2012).
Kiselycznyk, C. et al. NMDA receptor subunits and associated signaling molecules mediating antidepressant-related effects of NMDA-GluN2B antagonism. Behav. Brain Res. 287, 89–95 (2015).
Koike, H., Fukumoto, K., Iijima, M. & Chaki, S. Role of BDNF/TrkB signaling in antidepressant-like effects of a group II metabotropic glutamate receptor antagonist in animal models of depression. Behav. Brain Res. 238, 48–52 (2013).
Lang, E. et al. Molecular and cellular dissection of NMDA receptor subtypes as antidepressant targets. Neurosci. Biobehav. Rev. 84, 352–358 (2018).
Pothula, S. et al. Cell-type specific modulation of NMDA receptors triggers antidepressant actions. Mol. Psychiatry https://doi.org/10.1038/s41380-020-0796-3 (2020).
Campbell, S. & Macqueen, G. The role of the hippocampus in the pathophysiology of major depression. J. Psychiatry Neurosci. 29, 417–426 (2004).
Gu, X., Zhou, L. & Lu, W. An NMDA receptor-rependent mechanism underlies inhibitory synapse development. Cell Rep. 14, 471–478 (2016).
Tatard-Leitman, V. M. et al. Pyramidal cell selective ablation of N-methyl-d-aspartate receptor 1 causes increase in cellular and network excitability. Biol. Psychiatry 77, 556–568 (2015).
Kim, C. S., Chang, P. Y. & Johnston, D. Enhancement of dorsal hippocampal activity by knockdown of HCN1 channels leads to anxiolytic- and antidepressant-like behaviors. Neuron 75, 503–516 (2012).
Han, Y. et al. HCN-channel dendritic targeting requires bipartite interaction with TRIP8b and regulates antidepressant-like behavioral effects. Mol. Psychiatry 22, 458–465 (2017).
Kernell, D. Input resistance, electrical excitability, and size of ventral horn cells in cat spinal cord. Science 152, 1637–1640 (1966).
Hou, G. & Zhang, Z. W. NMDA receptors regulate the development of neuronal intrinsic excitability through cell-autonomous mechanisms. Front. Cell Neurosci. 11, 353 (2017).
Zanos, P. et al. NMDA receptor activation-dependent antidepressant-relevant behavioral and synaptic actions of ketamine. J. Neurosci. 43, 1038–1050 (2023).
Frizelle, P. A., Chen, P. E. & Wyllie, D. J. Equilibrium constants for (R)-[(S)-1-(4-bromo-phenyl)-ethylamino]-(2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-5-yl)-methyl]-phosphonic acid (NVP-AAM077) acting at recombinant NR1/NR2A and NR1/NR2B N-methyl-d-aspartate receptors: implications for studies of synaptic transmission. Mol. Pharmacol. 70, 1022–1032 (2006).
Russo, S. J. & Nestler, E. J. The brain reward circuitry in mood disorders. Nat. Rev. Neurosci. 14, 609–625 (2013).
Abdallah, C. G. et al. The nucleus accumbens and ketamine treatment in major depressive disorder. Neuropsychopharmacology 42, 1739–1746 (2017).
Khan, A. R. et al. Neurite atrophy in dorsal hippocampus of rat indicates incomplete recovery of chronic mild stress induced depression. NMR Biomed. 32, e4057 (2019).
Felix-Ortiz, A. C. & Tye, K. M. Amygdala inputs to the ventral hippocampus bidirectionally modulate social behavior. J. Neurosci. 34, 586–595 (2014).
Bagot, R. C. et al. Ventral hippocampal afferents to the nucleus accumbens regulate susceptibility to depression. Nat. Commun. 6, 7062 (2015).
Kim, C. S. & Johnston, D. Antidepressant effects of (S)-ketamine through a reduction of hyperpolarization-activated current Ih. iScience 23, 101239 (2020).
Homayoun, H. & Moghaddam, B. NMDA receptor hypofunction produces opposite effects on prefrontal cortex interneurons and pyramidal neurons. J. Neurosci. 27, 11496–11500 (2007).
Quirk, M. C., Sosulski, D. L., Feierstein, C. E., Uchida, N. & Mainen, Z. F. A defined network of fast-spiking interneurons in orbitofrontal cortex: responses to behavioral contingencies and ketamine administration. Front. Syst. Neurosci. 3, 13 (2009).
Picard, N., Takesian, A. E., Fagiolini, M. & Hensch, T. K. NMDA 2A receptors in parvalbumin cells mediate sex-specific rapid ketamine response on cortical activity. Mol. Psychiatry 24, 828–838 (2019).
Widman, A. J. & McMahon, L. L. Disinhibition of CA1 pyramidal cells by low-dose ketamine and other antagonists with rapid antidepressant efficacy. Proc. Natl Acad. Sci. USA 115, E3007–E3016 (2018).
Lu, W., Bushong, E. A., Shih, T. P., Ellisman, M. H. & Nicoll, R. A. The cell-autonomous role of excitatory synaptic transmission in the regulation of neuronal structure and function. Neuron 78, 433–439 (2013).
Kase, D. & Imoto, K. The role of HCN channels on membrane excitability in the nervous system. J. Signal Transduct. 2012, 619747 (2012).
Burrone, J., O’Byrne, M. & Murthy, V. N. Multiple forms of synaptic plasticity triggered by selective suppression of activity in individual neurons. Nature 420, 414–418 (2002).
Kadotani, H. et al. Motor discoordination results from combined gene disruption of the NMDA receptor NR2A and NR2C subunits, but not from single disruption of the NR2A or NR2C subunit. J. Neurosci. 16, 7859–7867 (1996).
Zhou, J. et al. NMDA receptor-dependent prostaglandin-endoperoxide synthase 2 induction in neurons promotes glial proliferation during brain development and injury. Cell Rep. 38, 110557 (2022).
Ruzankina, Y. et al. Deletion of the developmentally essential gene ATR in adult mice leads to age-related phenotypes and stem cell loss. Cell Stem Cell 1, 113–126 (2007).
Goebbels, S. et al. Genetic targeting of principal neurons in neocortex and hippocampus of NEX-Cre mice. Genesis 44, 611–621 (2006).
Vong, L. et al. Leptin action on GABAergic neurons prevents obesity and reduces inhibitory tone to POMC neurons. Neuron 71, 142–154 (2011).
Kaneko, R. et al. Inhibitory neuron-specific Cre-dependent red fluorescent labeling using VGAT BAC-based transgenic mouse lines with identified transgene integration sites. J. Comp. Neurol. 526, 373–396 (2018).
Madisen, L. et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).
Yang, Y. et al. Ketamine blocks bursting in the lateral habenula to rapidly relieve depression. Nature 554, 317–322 (2018).
Ryan, T. J., Roy, D. S., Pignatelli, M., Arons, A. & Tonegawa, S. Memory. Engram cells retain memory under retrograde amnesia. Science 348, 1007–1013 (2015).
Paxinos, G. & Franklin, K. B. J. The Mouse Brain In Stereotaxic Coordinates, 4th edn (Elsevier Science & Technology, 2013).
Zhang, Q., He, Q., Wang, J., Fu, C. & Hu, H. Use of TAI-FISH to visualize neural ensembles activated by multiple stimuli. Nat. Protoc. 13, 118–133 (2018).
Chen, Y. et al. Activity-induced Nr4a1 regulates spine density and distribution pattern of excitatory synapses in pyramidal neurons. Neuron 83, 431–443 (2014).
We thank S. Nakanishi (Osaka Bioscience Institute and Kyoto University, Japan) for providing the Grin2a−/− mice (by the RIKEN BRC through the National Bio-Resource Project of MEXT, Japan). We thank K. He (Interdisciplinary Research Center on Biology and Chemistry, Chinese Academy of Sciences, China), J. Hu (ShanghaiTech University, China), Z. Qiu (Institute of Neuroscience, Chinese Academy of Sciences, China) and X. Xu (Institute of Neuroscience, Chinese Academy of Sciences, China) for helping us to acquire the genetically engineered tool mice. We thank the staff members of the Animal Facility at the National Facility for Protein Science in Shanghai, Shanghai Advanced Research Institute and the Chinese Academy of Sciences for providing assistance in mouse breeding and maintenance. We thank M. Sheng (Broad Institute and Massachusetts Institute of Technology, USA) and J. Yuan (Interdisciplinary Research Center on Biology and Chemistry, Chinese Academy of Sciences, China) for helping with the manuscript preparation. The present study was supported by the following agencies: Shanghai Municipal Science and Technology Major Project to Y.C. (grant no. 2019SHZDZX02) and Natural Science Foundation of Shanghai to Y.G. (grant nos. 19ZR1468600 and 201409003800).
Y.C. is a visiting professor of Shanghai Jiao Tong University and a founder of Synphatec (Shanghai) Biopharmaceutical Technology Co., Ltd. Y.C., Y.G., T.S., Y.L. and C.F. are inventors on China patent application (no. 202110901806.8) held by Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences.
Peer review information
Nature Neuroscience thanks Gerard Sanacora, Panos Zanos and the other, anonymous, reviewer(s) 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 MK-801-induced fast antidepressant and anxiolytic effects, not hyperlocomotion, were occluded in GluN2A−/− mice, related to Fig. 1.
a and b, GluN2A expression was evaluated with FISH (a) and western blot (b) in cortex or hippocampus from WT and GluN2A−/− mice. Scale bar showed 200 μm. Samples of (b) derive from the same experiment and that gels/blots were processed in parallel. c–f, Measurements of GluN2A−/− mice and their WT littermates with OFT (c, p = 0.00020, d, p = 0.0040, WT n = 16, GluN2A−/− n = 14) and NSFT (e, p = 0.0031, f, WT n = 10, GluN2A−/− n = 11). g, Escape latency of WT and GluN2A−/− mice in learned helplessness test (LH) (WT-Sham n = 7, WT-IES n = 20, p = 0.0013; GluN2A−/−-Sham n = 10, GluN2A−/−-IES n = 19, p < 0.0001). h–j, Effects of MK-801 treatment on WT and GluN2A−/− mice were evaluated with OFT (WT-Sal n = 6, WT-MK-801 n = 8, GluN2A−/−-Sal n = 8, GluN2A−/−-MK-801 n = 9). h, WT-Sal v.s. WT-MK-801 p = 0.00040; WT-Sal v.s. GluN2A−/−-Sal p = 0.0080; GluN2A−/−-Sal v.s. GluN2A−/−- MK-801 p = 0.0010; i, WT-Sal v.s. WT-MK-801 p < 0.0001; WT-Sal v.s. GluN2A−/−-Sal p = 0.0080; GluN2A−/−-Sal v.s. GluN2A−/−-MK-801 p = 0.0019; j, WT-Sal v.s. WT-MK-801 p = 0.00010; WT-Sal v.s. GluN2A−/−-Sal p < 0.0001. k, Inhibition of GluN2A-containing NMDAR-mediated currents by ketamine, (2R, 6R)-HNK and LY341495. IComp/IGlu represents the normalized remaining currents after co-perfusion with each compound. Veh v.s. Ket p < 0.0001, one-way ANOVA. l, Schematic diagram showing the experimental timeline. m, Quantitation of the immobility time from WT and GluN2A−/− in TST after 7 days fluoxetine (Flx) treatment (WT-Sal n = 10, WT-Flx n = 12, p = 0.0046; GluN2A−/−-Sal n = 9, GluN2A−/−-Flx n = 12, p = 0.030; WT-Sal v.s. GluN2A−/−-Sal p = 0.0039). Error bars show SEM. Two-way ANOVA (g, h, i, j, m) and Student’s t test (two-tailed) (c–f). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Extended Data Fig. 2 Validation of adult age-specific induced GluN2A knockout in mice, related to Fig. 2.
a, Body weight of UBC-CreERT/Grin2aflox/flox mice treated with TAM (n = 22) or Veh (n = 20). b–d, GluN2A expression level was measured from hippocampus of UBC-CreERT/Grin2aflox/flox mice after treated with TAM or Veh (b, c, western blot, Veh n = 7, TAM n = 6) (d, qRT-PCR, Veh n = 5, TAM n = 5). c, p < 0.0001; d, p < 0.0001; e–i, Evaluation of Grin2aflox/flox (2A flox) and UBC-CreERT/Grin2aflox/flox (UBC-2A flox) mice with OFT (e), EPM (f, g), TST (h, p = 0.011) or FST (i, p = 0.025). The behavioral analysis was done 1 month after treatment with TAM (2A flox n = 8, UBC-2A flox n = 9). j and k, Verification of GluN2A knockout in UBC-CreERT/Grin2aflox/flox mice treated with TAM after inescapable footshocks training in LH test presented in Fig. 2i–n (Veh n = 5, TAM n = 6). k, p < 0.0001. Samples of (b)/(j) derive from the same experiment and that gels/blots were processed in parallel. Error bars show SEM. One-sample t test (c, k), two-way ANOVA (e–g) and Student’s t test (two-tailed) (d, h, i). * p < 0.05, **** p < 0.0001.
Extended Data Fig. 3 Validation of excitatory-neuron specific GluN2A conditional knockout mice (Nex-2A cKO), related to Fig. 3.
a, Greyscale images showing tdTomato signal in sagittal brain sections of Nex-Cre/Ai9 mice. C-cortex, H-hippocampus, S-striatum, T-thalamus, A-amygdala, M-midbrain. Scale bars showed 500 μm (top) and 200 μm (bottom) respectively. b and c, Representative greyscale images of Grin2a FISH in hippocampus (b) or cortex (c) of WT and Nex-2A cKO mice. Scale bar showed 50 μm in b and 100 μm in c. d and e, Western blot analysis of GluN2A in hippocampus (d) and cortex (e) of Nex-2A WT and cKO mice. Samples of (d)/(e) derive from the same experiment and that gels/blots were processed in parallel. f and g, Quantification of GluN2A expression level by western blot (f) and RT-PCR (g) in hippocampus or cortex of Nex-2A WT and cKO mice (f, Hipp: Nex-2A WT n = 4, cKO n = 5, p < 0.0001; Ctx: Nex-2A WT n = 4, cKO n = 3, p = 0.010), (g, Hipp: Nex-2A WT n = 4, cKO n = 3, p = 0.0045; Ctx: Nex-2A WT n = 4, cKO n = 3, p = 0.00020). One-sample t test (f) and Student’s t test (two-tailed) (g). Error bars show SEM. ** p < 0.01, *** p < 0.001, **** p < 0.0001.
Extended Data Fig. 4 Validation of inhibitory-neuron specific GluN2A conditional knockout mice (vGAT-2A cKO), related to Fig. 3.
a, Greyscale images showing tdTomato signal in sagittal brain sections from vGAT-Cre/Ai9 mice. C-cortex, H-hippocampus, S-striatum, T-thalamus, A-amygdala, M-midbrain. Scale bars showed 500 μm (top) and 200 μm (bottom), respectively. b and c, Western blot analysis of GluN2A expression level in hippocampus of vGAT-2A cKO or WT mice, quantified in b (vGAT-2A WT n = 3, vGAT-2A cKO n = 3). Samples of (b) derive from the same experiment and that gels/blots were processed in parallel. d, Quantification of GluN2A mRNA level in hippocampus of vGAT-2A cKO or WT mice (vGAT-2A WT n = 4, vGAT-2A cKO n = 6). e–g, Grin2a FISH and fluorescent staining of GAD1 in hippocampus of vGAT-2A cKO or WT mice (e; yellow arrows indicated the GAD1 positive cells that co-expressed with GluN2A; cyan arrowhead indicated GAD1 positive cells without GluN2A signal). f, Area of stratum radiatum (SR) and stratum lacunosum & moleculare (SLM). g, Density of GluN2A positive cells in SR and SLM (vGAT-2A WT n = 4, vGAT-2A cKO n = 4; p = 0.0024,). Scale bar showed 200 μm. h, Representative traces of evoked NMDAR-mediated EPSCs from CA1 pyramidal neurons or interneurons in WT or vGAT-2A cKO mice after incubated with Veh (black) or GluN2B selective inhibitor Ro-25 6981 (blue). i and j, Quantitation of the decay time (i) or +Ro25/-Ro25 ratio of the amplitudes of evoked NMDAR-EPSCs (j) (Pyr: pyramidal neurons, vGAT-2A WT n = 3, vGAT-2A cKO n = 5; INs: interneurons, vGAT-2A WT n = 5, vGAT-2A cKO n = 5;). i, INs: p = 0.018; j, INs: p = 0.012. Error bars show SEM. One-sample t test (c, d), two-way ANOVA (f, i, j) and Student’s t test (two-tailed) (g).* p < 0.05, ** p < 0.01.
Extended Data Fig. 5 Hippocampal interneurons of GluN2A−/− mice had normal excitability, related to Fig. 6.
a–j, Representative current-clamp recording traces after injection of a 330 pA depolarizing current (a, f), quantitation of AP number (b, g), RMP (c, h), sample traces after step hyperpolarizing current injections (d, i), current-voltage curves and RIn (e, j and inserts) from pyramidal neurons of mPFC (a–e) or visual cortex (f–j) of WT (grey) or GluN2A−/− (red) mice. (mPFC: WT n = 9, GluN2A−/− n = 16; visual cortex: WT n = 13, GluN2A−/− n = 9). b, p(130 pA) = 0.012, p(150 pA) = 0.018; e, p = 0.031, p(RIn) = 0.031. k–p, Representative current-clamp recording traces after injection of a 330 (k) or 210 (n) pA depolarizing current (k, n), quantitation of AP number after depolarizing current injections (l, o), RMP (m, p) from non-fast spiking (k–m) or fast-spiking (n–p) interneurons from CA1 of WT (grey) or GluN2A−/− (red) mice. (Non-fast, WT n = 12, GluN2A−/− n = 7; Fast, WT n = 13, GluN2A−/− n = 14). 6–8 weeks old mice were used for electrophysiological recordings. Error bars show SEM. Two-way ANOVA (b, e, g, j, l, o) or Student’s t test (two-tailed) (c, e (insert), h, j (insert), m, p). * p < 0.05.
Extended Data Fig. 6 Cell type specific removal of GluN2A did not alter excitatory or inhibitory synaptic transmission of CA1 pyramidal neurons, related to Fig. 6.
a–f, Representative traces (a, d), average individual events (b, e), quantitation of cumulative probability and average amplitude of events (c, f) of mIPSCs and sIPSCs recorded from 4–6 weeks old Nex-2A WT (black) or Nex-2A cKO (olive) mice (mIPSCs: Nex-2A WT n = 12, Nex-2A cKO n = 14; sIPSCs: Nex-2A WT n = 16, Nex-2A cKO n = 14). g–r, Representative traces, average individual events, quantitation of cumulative probability and average amplitude of events of mIPSCs (g–i), sIPSCs (j–l), mEPSCs (m–o) and sEPSCs (p–r) recorded from vGAT-2A WT (black) or vGAT-2A cKO (purple) mice. (mIPSCs: vGAT-2A WT n = 18, vGAT-2A cKO n = 21; sIPSCs: vGAT-2A WT n = 15, vGAT-2A cKO n = 13; mEPSCs: vGAT-2A WT n = 13, vGAT-2A cKO n = 10; sEPSCs: vGAT-2A WT n = 12, vGAT-2A cKO n = 16) 6–8 weeks mcie were used for electrophysiological recordings. Error bars showed SEM. Cumulative frequency was analyzed with Kolmogorov-Smirnov test. Peak amplitudes were analyzed with Student’s t test (two-tailed).
Extended Data Fig. 7 GluN2B was not responsible to the effects of MK-801 in intrinsic excitability and antidepressant-like behaviors, related to Fig. 6.
a, Representative recording traces after injection of 330 pA depolarizing current in CA1 pyramidal neurons treated with Veh or 3 μM Ro-25 6981 for 1–2 hours. b and c, Quantitation of the number of action potentials (b) after injection of depolarizing currents and RMP (c) recorded from neurons after Veh or Ro-25 6981 treatments (Veh n = 18, Ro 25-6981 n = 19). d–i, number of AP (e, p = 0.0074), RMP (f, p = 0.041), current-voltage curve (h, p = 0.00070), input resistance (RIn) (i, p = 0.00080) and sample trace recorded from CA1 pyramidal neurons after incubated with ‘Ro 25-6981 + Veh’ (n = 10) or ‘Ro 25-6981 + 10 μM MK-801’ (n = 10) for 1–2 hours from WT mice (g). j and k, Effects of MK-801 on WT mice-treated with GluN2B inhibitor Ro 25-6981 (10 mpk) were evaluated with TST (j; Ro 25-6981 + Sal n = 10; Ro 25-6981 + MK-801 n = 10, p < 0.0001) and FST (k; Ro 25-6981 + Sal n = 10; Ro 25-6981 + MK-801 n = 9, p < 0.0001). All the recordings were measured from CA1 pyramidal neurons acutely prepared from 6-8 weeks old WT mice. Error bars show SEM. Two-way ANOVA (b, e, h) or Student’s t test (two-tailed) (c, f, i, j, k). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
a, Representative voltage sag were generated by hyperpolarized current of −100 pA. b, Normalized voltage sag of WT and GluN2A−/−, which were measured at the last 50-ms of 1-s stimulation (WT n = 14, GluN2A−/− n = 22). Error bars show SEM. Two-way ANOVA.
Unprocessed western blots.
Unprocessed western blots.
Unprocessed western blots.
Unprocessed western blots.
About this article
Cite this article
Su, T., Lu, Y., Fu, C. et al. GluN2A mediates ketamine-induced rapid antidepressant-like responses. Nat Neurosci 26, 1751–1761 (2023). https://doi.org/10.1038/s41593-023-01436-y