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Cell-Type-Specific Role for Nucleus Accumbens Neuroligin-2 in Depression and Stress Susceptibility

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Cell-Type-Specific Role for Nucleus Accumbens Neuroligin-2 in Depression and Stress Susceptibility Cell-type-specific role for nucleus accumbens neuroligin-2 in depression and stress susceptibility Mitra Heshmatia,b, Hossein Aleyasina,b, Caroline Menarda,b, Daniel J. Christoffelc, Meghan E. Flanigana,b, Madeline L. Pfaua,b, Georgia E. Hodesa,b,1, Ashley E. Lepacka,b, Lucy K. Bicksa,b, Aki Takahashia,b,d, Ramesh Chandrae, Gustavo Tureckif, Mary Kay Loboe, Ian Mazea,b, Sam A. Goldeng, and Scott J. Russoa,b,2 aFishberg Department of Neuroscience, Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029; bFriedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029; cNancy Pritzker Laboratory, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305; dLaboratory of Behavioral Neuroendocrinology, University of Tsukuba, Ibaraki 305-8577, Japan; eDepartment of Anatomy and Neurobiology, University of Maryland School of Medicine, Baltimore, MD 21201; fMcGill Group for Suicide Studies, Douglas Mental Health University Institute, McGill University, Montreal, QC H3A 0G4, Canada; and gBehavioral Neuroscience Branch, Intramural Research Program, NIDA-NIH, Baltimore, MD 21224 Edited by Bruce S. McEwen, The Rockefeller University, New York, NY, and approved December 19, 2017 (received for review October 31, 2017) Behavioral coping strategies are critical for active resilience to stress NAc contains primarily GABAergic medium spiny neurons and depression; here we describe a role for neuroligin-2 (NLGN-2) in (MSNs) divided into two subpopulations that often have distinct the nucleus accumbens (NAc). Neuroligins (NLGN) are a family of functional effects on stress- and reward-mediated behaviors: do- neuronal postsynaptic cell adhesion proteins that are constituents pamine receptor 1-positive medium spiny neurons (D1-MSNs), of the excitatory and inhibitory synapse. Importantly, NLGN-3 and expressing primarily dopamine D1 receptors, and D2-MSNs, NLGN-4 mutations are strongly implicated as candidates underlying expressing primarily dopamine D2 receptors (27, 28, 31, 32). the development of neuropsychiatric disorders with social distur- Two recent studies have examined NLGN-2 expression in striatal bances such as autism, but the role of NLGN-2 in neuropsychiatric MSNs (33, 34), confirming its role as a constituent of the MSN disease states is unclear. Here we show a reduction in NLGN-2 gene inhibitory synapse and quantifying NLGN-2 expression within expression in the NAc of patients with major depressive disorder. MSN populations. Here, we provide evidence that NLGN-2 is Chronic social defeat stress in mice also decreases NLGN-2 selectively reduced in the NAc of patients with MDD and describe a cell- NEUROSCIENCE in dopamine D1-positive cells, but not dopamine D2-positive cells, type-specific role for NLGN-2 in NAc MSNs in modulating active within the NAc of stress-susceptible mice. Functional NLGN-2 knock- behavioral coping mechanisms in stress susceptibility and resil- down produces bidirectional, cell-type-specific effects: knockdown ience in a mouse model of depression. in dopamine D1-positive cells promotes subordination and stress susceptibility, whereas knockdown in dopamine D2-positive cells Results mediates active defensive behavior. These findings establish a be- NLGN-2 Decreases in NAc of Depressed Patients. Transcriptional havioral role for NAc NLGN-2 in stress and depression; provide a profiling of postmortem human NAc was conducted to identify basis for targeted, cell-type specific therapy; and highlight the role changes in neuroligin gene expression in MDD, revealing a se- of active behavioral coping mechanisms in stress susceptibility. lective down-regulation of NLGN-2 in subjects with MDD that is not significantly reversed by antidepressant medication being neuroligin-2 | depression | social defeat stress | dominance | medium spiny neuron Significance ehavioral coping strategies used during stressful events play Ban important role in determining the subsequent develop- Although mutations in the neuroligin-3 and neuroligin-4 genes ment of neuropsychiatric disease (1, 2). This same phenomenon is are implicated in autism syndromes, very little is known about observed in rodent stress models, where susceptibility or resilience the contribution of neuroligin-2 to neuropsychiatric disease to social stress is strongly influenced by the subordinate’sselection states. We report a decrease in neuroligin-2 gene expression of either submissive or dominant coping styles (3–5). However, the in the postmortem nucleus accumbens (NAc) of depressed underlying synaptic mechanisms promoting the selection of such patients. Reverse translation of this finding in chronic social coping behaviors are unclear (6) and present a therapeutic ap- defeat stress, an animal model of depression that enables in- proach to the treatment of stress-related disorders. vestigation of both susceptibility and resiliency mechanisms, Neuroligin-2 (NLGN-2) is a structural constituent of the in- uncovers an important functional role for NAc neuroligin-2 in hibitory synapse (7, 8), and although it is also associated with stress susceptibility. We detail a cell-type-specific role for NAc maladaptive social behavior (9–13), its direct role in neuropsy- neuroligin-2 in modulating social avoidance behavior and chiatric disease is unclear (14, 15). NLGN-2 is a key postsynaptic dominance behaviors important for resiliency. Together, these cell adhesion protein that supports the functional integrity of the findings describe a role for NAc neuroligin-2 in depression and inhibitory synapse (7, 14, 16). Studies of developmental NLGN- chronic stress behaviors. 2 knockout mice have established a distinct role for NLGN-2 in Author contributions: M.H., S.A.G., and S.J.R. designed research; M.H., H.A., C.M., D.J.C., maintaining inhibitory synapse function and modulating anxiety M.E.F., M.L.P., G.E.H., A.E.L., L.K.B., A.T., R.C., and S.A.G. performed research; G.T., M.K.L., behaviors (8, 17–19). Outside of this developmental window, and I.M. contributed new reagents/analytic tools; M.H., H.A., C.M., D.J.C., R.C., S.A.G., and very little is known about the role of NLGN-2 in regulating adult S.J.R. analyzed data; and M.H., S.A.G., and S.J.R. wrote the paper. social behavior. Recent NLGN-2 manipulations in the adult mouse The authors declare no conflict of interest. prefrontal cortex and hippocampus describe varying effects on This article is a PNAS Direct Submission. anxiety, aggression, and fear memory (9, 20–22). Published under the PNAS license. The NAc, an integral hub in the mesolimbic dopamine circuit, 1Present address: Department of Neuroscience, Virginia Polytechnic Institute and State is well established as a critical brain region regulating social University, Blacksburg, VA 24060. stress behaviors (23). Synaptic plasticity within the dopaminergic 2To whom correspondence should be addressed. Email: [email protected]. mesolimbic circuit is implicated in both major depressive disor- This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. der (MDD) and increased susceptibility to stress (24–30). The 1073/pnas.1719014115/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1719014115 PNAS Early Edition | 1of6 Downloaded by guest on September 25, 2021 ABControl (14) 2.0 CD2.0 6.0 Depressed (13) Medicated (16) * 1.5 1.5 4.5 1.0 1.0 3.0 Nlgn1 Nlgn2 Nlgn3 0.5 0.5 1.5 mRNA fold change fold mRNA mRNA fold change fold mRNA change fold mRNA 0.0 0.0 0.0 Fig. 1. NLGN-2 in the human postmortem nucleus accumbens changes in MDD. (A) Schematic indicating blue region of dissection of postmortem human NAc. Transcriptional profiling of postmortem human NAc (B) NLGN-1, (C)NLGN-2,and(D)NLGN-3geneexpressionrevealedaselectivedecreaseinNLGN-2indepressed subjects (one-way ANOVA: F2,42 = 3.876; *P < 0.05). All data are represented as mean ± SEM. See SI Appendix,TableS1for detailed demographic data. present at the time of death (Fig. 1 and SI Appendix,TableS1for NAc NLGN-2 only in mice that are susceptible to stress (Fig. 2). demographics). The change in NAc NLGN-2 does not generalize After stress, mice are classified as susceptible or resilient based on to other neuropsychiatric disorders such as chronic cocaine abuse social interaction (35–37) (SI Appendix, Fig. S2). We report a (SI Appendix, Fig. S1). This is evidence for neuroligin gene regu- decrease in NAc NLGN-2 protein that significantly correlates with lation in the NAc in MDD and presents a unique role for NLGN- lower social interaction ratio, or increased susceptibility to stress 2 in neuropsychiatric disease and antidepressant strategies. (Fig. 2 B–D). To characterize the cell-type specificity of social- defeat-induced changes, NLGN-2 mRNA was assayed in the NLGN-2 Decreases in NAc of Stress-Susceptible Mice. Reverse trans- principal NAc neuronal populations, D1-MSNs and D2-MSNs, lation of these findings to chronic social defeat stress, a mouse using RiboTag methodology (32). NLGN-2 mRNA levels de- model of depression-like behavior, recapitulates a decrease in creased specifically in D1-MSNs of stress-susceptible mice, and A B1 B2 B3 Physical Sensory (10 min) (24 hours) 10x Control Susceptible Resilient C 0.20 Control (7) D 0.20 **p =0.0006 2 Susceptible (6) Resilient (9) 0.15 0.15 0.10 0.10 0.05 * 0.05 Social Neuroligin-2 puncta / / puncta Neuroligin-2 m2 Neuroligin-2 puncta / / puncta Neuroligin-2 m Interaction 0.00 0.00 0.0 1.0 2.0 SI Ratio EFGHControl (7) Control (6) 1.5 2.0 1.5 2.0 Susceptible (7) *p = 0.04 SusceptibleSusceptible (8)(8 p = 0.98 Resilient (4) ResilientResilient (4)(4) 1.5 1.5 1.0 1.0 nlgn2 nlgn2 nlgn2 nlgn2 nlgn2 nlgn2 nlgn2 nlgn2 * 1.0 1.0 D2 - MSN D2 - MSN D1 - MSN 0.5 D1 - MSN 0.5 0.5 0.5 Fold change Fold change Fold change Fold change 0.0 0.0 0.0 0.0 01234 01234 SI Ratio SI Ratio Fig.
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