Increased excitatory amino acid transporter 2 levels in basolateral amygdala astrocytes mediate chronic stress-induced anxiety-like behavior.

Publisher: Wolters Kluwer Health, Medknow Country of Publication: India NLM ID: 101316351 Publication Model: Print-Electronic Cited Medium: Print ISSN: 1673-5374 (Print) Linking ISSN: 16735374 NLM ISO Abbreviation: Neural Regen Res Subsets: PubMed not MEDLINE

Publication: 2013 - : Mumbai : Wolters Kluwer Health, Medknow
Original Publication: Shenyang : Editorial Board of Neural Regeneration Research

JOURNAL/nrgr/04.03/01300535-202506000-00024/figure1/v/2024-08-05T133530Z/r/image-tiff The conventional perception of astrocytes as mere supportive cells within the brain has recently been called into question by empirical evidence, which has revealed their active involvement in regulating brain function and encoding behaviors associated with emotions. Specifically, astrocytes in the basolateral amygdala have been found to play a role in the modulation of anxiety-like behaviors triggered by chronic stress. Nevertheless, the precise molecular mechanisms by which basolateral amygdala astrocytes regulate chronic stress-induced anxiety-like behaviors remain to be fully elucidated. In this study, we found that in a mouse model of anxiety triggered by unpredictable chronic mild stress, the expression of excitatory amino acid transporter 2 was upregulated in the basolateral amygdala. Interestingly, our findings indicate that the targeted knockdown of excitatory amino acid transporter 2 specifically within the basolateral amygdala astrocytes was able to rescue the anxiety-like behavior in mice subjected to stress. Furthermore, we found that the overexpression of excitatory amino acid transporter 2 in the basolateral amygdala, whether achieved through intracranial administration of excitatory amino acid transporter 2 agonists or through injection of excitatory amino acid transporter 2-overexpressing viruses with GfaABC1D promoters, evoked anxiety-like behavior in mice. Our single-nucleus RNA sequencing analysis further confirmed that chronic stress induced an upregulation of excitatory amino acid transporter 2 specifically in astrocytes in the basolateral amygdala. Moreover, through in vivo calcium signal recordings, we found that the frequency of calcium activity in the basolateral amygdala of mice subjected to chronic stress was higher compared with normal mice. After knocking down the expression of excitatory amino acid transporter 2 in the basolateral amygdala, the frequency of calcium activity was not significantly increased, and anxiety-like behavior was obviously mitigated. Additionally, administration of an excitatory amino acid transporter 2 inhibitor in the basolateral amygdala yielded a notable reduction in anxiety level among mice subjected to stress. These results suggest that basolateral amygdala astrocytic excitatory amino acid transporter 2 plays a role in in the regulation of unpredictable chronic mild stress-induced anxiety-like behavior by impacting the activity of local glutamatergic neurons, and targeting excitatory amino acid transporter 2 in the basolateral amygdala holds therapeutic promise for addressing anxiety disorders.
(Copyright © 2025 Copyright: © 2025 Neural Regeneration Research.)

Alborghetti M, Bianchini E, De Carolis L, Galli S, Pontieri FE, Rinaldi D (2024) Type-B monoamine oxidase inhibitors in neurological diseases: clinical applications based on preclinical findings. Neural Regen Res 19:16–21.
Alijanpour S, Miryounesi M, Ghafouri-Fard S (2023) The role of excitatory amino acid transporter 2 (EAAT2) in epilepsy and other neurological disorders. Metab Brain Dis 38:1–16.
Allen NJ, Eroglu C (2017) Cell biology of astrocyte-synapse interactions. Neuron 96:697–708.
Autry AE, Grillo CA, Piroli GG, Rothstein JD, McEwen BS, Reagan LP (2006) Glucocorticoid regulation of GLT-1 glutamate transporter isoform expression in the rat hippocampus. Neuroendocrinology 83:371–379.
Azarias G, Perreten H, Lengacher S, Poburko D, Demaurex N, Magistretti PJ, Chatton JY (2011) Glutamate transport decreases mitochondrial pH and modulates oxidative metabolism in astrocytes. J Neurosci 31:3550–3559.
Bjørnsen LP, Hadera MG, Zhou Y, Danbolt NC, Sonnewald U (2014) The GLT-1 (EAAT2; slc1a2) glutamate transporter is essential for glutamate homeostasis in the neocortex of the mouse. J Neurochem 128:641–649.
Bourin M, Hascoët M (2003) The mouse light/dark box test. Eur J Pharmacol 463:55–65.
Brandebura AN, Paumier A, Onur TS, Allen NJ (2023) Astrocyte contribution to dysfunction, risk and progression in neurodegenerative disorders. Nat Rev Neurosci 24:23–39.
Cai W, Xue C, Sakaguchi M, Konishi M, Shirazian A, Ferris HA, Li ME, Yu R, Kleinridders A, Pothos EN, Kahn CR (2018) Insulin regulates astrocyte gliotransmission and modulates behavior. J Clin Invest 128:2914–2926.
Chattarji S, Tomar A, Suvrathan A, Ghosh S, Rahman MM (2015) Neighborhood matters: divergent patterns of stress-induced plasticity across the brain. Nat Neurosci 18:1364–1375.
Chatton JY, Magistretti PJ, Barros LF (2016) Sodium signaling and astrocyte energy metabolism. Glia 64:1667–1676.
Cho WH, Noh K, Lee BH, Barcelon E, Jun SB, Park HY, Lee SJ (2022) Hippocampal astrocytes modulate anxiety-like behavior. Nat Commun 13:6536.
COVID-19 Mental Disorders Collaborators (2021) Global prevalence and burden of depressive and anxiety disorders in 204 countries and territories in 2020 due to the COVID-19 pandemic. Lancet 398:1700–1712.
D’Egidio F, Castelli V, Lombardozzi G, Ammannito F, Cimini A, d’Angelo M (2024) Therapeutic advances in neural regeneration for Huntington’s disease. Neural Regen Res 19:1991–1997.
Fan J, Guo F, Mo R, Chen LY, Mo JW, Lu CL, Ren J, Zhong QL, Kuang XJ, Wen YL, Gu TT, Liu JM, Li SJ, Fang YY, Zhao C, Gao TM, Cao X (2023) O-GlcNAc transferase in astrocytes modulates depression-related stress susceptibility through glutamatergic synaptic transmission. J Clin Invest 133:e160016.
Fu X, Teboul E, Weiss GL, Antonoudiou P, Borkar CD, Fadok JP, Maguire J, Tasker JG (2022) Gq neuromodulation of BLA parvalbumin interneurons induces burst firing and mediates fear-associated network and behavioral state transition in mice. Nat Commun 13:1290.
Gasull-Camós J, Tarrés-Gatius M, Artigas F, Castañé A (2017) Glial GLT-1 blockade in infralimbic cortex as a new strategy to evoke rapid antidepressant-like effects in rats. Transl Psychiatry 7:e1038.
Gomez JA, Perkins JM, Beaudoin GM, Cook NB, Quraishi SA, Szoeke EA, Thangamani K, Tschumi CW, Wanat MJ, Maroof AM, Beckstead MJ, Rosenberg PA, Paladini CA (2019) Ventral tegmental area astrocytes orchestrate avoidance and approach behavior. Nat Commun 10:1455.
Green JL, Dos Santos WF, Fontana ACK (2021) Role of glutamate excitotoxicity and glutamate transporter EAAT2 in epilepsy: Opportunities for novel therapeutics development. Biochem Pharmacol 193:114786.
Gross C, Hen R (2004) The developmental origins of anxiety. Nat Rev Neurosci 5:545–552.
Hintiryan H, et al. (2021) Connectivity characterization of the mouse basolateral amygdalar complex. Nat Commun 12:2859.
Iovino L, et al. (2022) Trafficking of the glutamate transporter is impaired in LRRK2-related Parkinson’s disease. Acta Neuropathol 144:81–106.
Jimenez JC, Su K, Goldberg AR, Luna VM, Biane JS, Ordek G, Zhou P, Ong SK, Wright MA, Zweifel L, Paninski L, Hen R, Kheirbek MA (2018) Anxiety cells in a hippocampal-hypothalamic circuit. Neuron 97:670–683.e6.
Kruyer A, Angelis A, Garcia-Keller C, Li H, Kalivas PW (2022) Plasticity in astrocyte subpopulations regulates heroin relapse. Sci Adv 8:eabo7044.
Lee HG, Wheeler MA, Quintana FJ (2022) Function and therapeutic value of astrocytes in neurological diseases. Nat Rev Drug Discov 21:339–358.
Lee Y, Gaskins D, Anand A, Shekhar A (2007) Glia mechanisms in mood regulation: a novel model of mood disorders. Psychopharmacology (Berl) 191:55–65.
Li KX, Lu M, Cui MX, Wang XM, Zheng Y (2023) Astrocyte-neuron communication mediated by the Notch signaling pathway: focusing on glutamate transport and synaptic plasticity. Neural Regen Res 18:2285–2290.
Li S, Cao W, Zhou S, Ma M, Zhang W, Li F, Li C (2021a) Expression of Cntn1 is regulated by stress and associated with anxiety and depression phenotypes. Brain Behav Immun 95:142–153.
Li X, Zhong H, Wang Z, Xiao R, Antonson P, Liu T, Wu C, Zou J, Wang L, Nalvarte I, Xu H, Warner M, Gustafsson JA, Fan X (2021b) Loss of liver X receptor β in astrocytes leads to anxiety-like behaviors via regulating synaptic transmission in the medial prefrontal cortex in mice. Mol Psychiatry 26:6380–6393.
Lin CL, Kong Q, Cuny GD, Glicksman MA (2012) Glutamate transporter EAAT2: a new target for the treatment of neurodegenerative diseases. Future Med Chem 4:1689–1700.
Liu WZ, Zhang WH, Zheng ZH, Zou JX, Liu XX, Huang SH, You WJ, He Y, Zhang JY, Wang XD, Pan BX (2020) Identification of a prefrontal cortex-to-amygdala pathway for chronic stress-induced anxiety. Nat Commun 11:2221.
Lovick TA, Zangrossi H Jr. (2021) Effect of estrous cycle on behavior of females in rodent tests of anxiety. Front Psychiatry 12:711065.
Luo ZY, Huang L, Lin S, Yin YN, Jie W, Hu NY, Hu YY, Guan YF, Liu JH, You QL, Chen YH, Luo ZC, Zhang SR, Li XW, Yang JM, Tao YM, Mei L, Gao TM (2020) Erbin in amygdala parvalbumin-positive neurons modulates anxiety-like behaviors. Biol Psychiatry 87:926–936.
Magi S, Piccirillo S, Amoroso S, Lariccia V (2019) Excitatory amino acid transporters (EAATs): glutamate transport and beyond. Int J Mol Sci 20:5674.
Martin-Fernandez M, Jamison S, Robin LM, Zhao Z, Martin ED, Aguilar J, Benneyworth MA, Marsicano G, Araque A (2017) Synapse-specific astrocyte gating of amygdala-related behavior. Nat Neurosci 20:1540–1548.
Masneuf S, Lowery-Gionta E, Colacicco G, Pleil KE, Li C, Crowley N, Flynn S, Holmes A, Kash T (2014) Glutamatergic mechanisms associated with stress-induced amygdala excitability and anxiety-related behavior. Neuropharmacology 85:190–197.
McEwen BS, Bowles NP, Gray JD, Hill MN, Hunter RG, Karatsoreos IN, Nasca C (2015) Mechanisms of stress in the brain. Nat Neurosci 18:1353–1363.
McNeill J, Rudyk C, Hildebrand ME, Salmaso N (2021) Ion channels and electrophysiological properties of astrocytes: implications for emergent stimulation technologies. Front Cell Neurosci 15:644126.
Murphy-Royal C, Gordon GR, Bains JS (2019) Stress-induced structural and functional modifications of astrocytes-Further implicating glia in the central response to stress. Glia 67:1806–1820.
Naour AL, Beziat E, Kam JH, Magistretti P, Benabid AL, Mitrofanis J (2023) Do astrocytes respond to light, sound, or electrical stimulation? Neural Regen Res 18:2343–2347.
Olloquequi J, Cornejo-Córdova E, Verdaguer E, Soriano FX, Binvignat O, Auladell C, Camins A (2018) Excitotoxicity in the pathogenesis of neurological and psychiatric disorders: Therapeutic implications. J Psychopharmacol 32:265–275.
Pál B (2024) On the functions of astrocyte-mediated neuronal slow inward currents. Neural Regen Res 19:2602–2612.
Pajarillo E, Rizor A, Lee J, Aschner M, Lee E (2019) The role of astrocytic glutamate transporters GLT-1 and GLAST in neurological disorders: Potential targets for neurotherapeutics. Neuropharmacology 161:107559.
Parkin GM, Udawela M, Gibbons A, Dean B (2018) Glutamate transporters, EAAT1 and EAAT2, are potentially important in the pathophysiology and treatment of schizophrenia and affective disorders. World J Psychiatry 8:51–63.
Percie du Sert N, et al. (2020) The ARRIVE guidelines 2.0: updated guidelines for reporting animal research. PLoS Biol 18:e3000410.
Perkins EM, Clarkson YL, Suminaite D, Lyndon AR, Tanaka K, Rothstein JD, Skehel PA, Wyllie DJA, Jackson M (2018) Loss of cerebellar glutamate transporters EAAT4 and GLAST differentially affects the spontaneous firing pattern and survival of Purkinje cells. Hum Mol Genet 27:2614–2627.
Popoli M, Yan Z, McEwen BS, Sanacora G (2011) The stressed synapse: the impact of stress and glucocorticoids on glutamate transmission. Nat Rev Neurosci 13:22–37.
Qiu B, Boudker O (2023) Symport and antiport mechanisms of human glutamate transporters. Nat Commun 14:2579.
Reagan LP, Rosell DR, Wood GE, Spedding M, Muñoz C, Rothstein J, McEwen BS (2004) Chronic restraint stress up-regulates GLT-1 mRNA and protein expression in the rat hippocampus: reversal by tianeptine. Proc Natl Acad Sci U S A 101:2179–2184.
Roozendaal B, McEwen BS, Chattarji S (2009) Stress, memory and the amygdala. Nat Rev Neurosci 10:423–433.
Sanacora G, Yan Z, Popoli M (2022) The stressed synapse 2.0: pathophysiological mechanisms in stress-related neuropsychiatric disorders. Nat Rev Neurosci 23:86–103.
Shao LX, Jiang Q, Liu XX, Gong DM, Yin YX, Wu G, Sun NH, Wang CK, Chen QZ, Yu C, Shi WX, Fan HY, Fukunaga K, Chen Z, Lu YM, Han F (2019) Functional coupling of Tmem74 and HCN1 channels regulates anxiety-like behavior in BLA neurons. Mol Psychiatry 24:1461–1477.
Sharma A, Kazim SF, Larson CS, Ramakrishnan A, Gray JD, McEwen BS, Rosenberg PA, Shen L, Pereira AC (2019) Divergent roles of astrocytic versus neuronal EAAT2 deficiency on cognition and overlap with aging and Alzheimer’s molecular signatures. Proc Natl Acad Sci U S A 116:21800–21811.
Stein MB, Simmons AN, Feinstein JS, Paulus MP (2007) Increased amygdala and insula activation during emotion processing in anxiety-prone subjects. Am J Psychiatry 164:318–327.
Suslova M, Kortzak D, Machtens JP, Kovermann P, Fahlke C (2023) Apo state pore opening as functional basis of increased EAAT anion channel activity in episodic ataxia 6. Front Physiol 14:1147216.
Takaba R, Ibi D, Watanabe K, Hayakawa K, Nakasai G, Hiramatsu M (2022) Role of sirtuin1 in impairments of emotion-related behaviors in mice with chronic mild unpredictable stress during adolescence. Physiol Behav 257:113971.
Takahashi K, Foster JB, Lin CL (2015) Glutamate transporter EAAT2: regulation, function, and potential as a therapeutic target for neurological and psychiatric disease. Cell Mol Life Sci 72:3489–3506.
Timper K, Del Río-Martín A, Cremer AL, Bremser S, Alber J, Giavalisco P, Varela L, Heilinger C, Nolte H, Trifunovic A, Horvath TL, Kloppenburg P, Backes H, Brüning JC (2020) GLP-1 receptor signaling in astrocytes regulates fatty acid oxidation, mitochondrial integrity, and function. Cell Metab 31:1189–1205.e13.
Tovote P, Fadok JP, Lüthi A (2015) Neuronal circuits for fear and anxiety. Nat Rev Neurosci 16:317–331.
Tsai SF, Hsu PL, Chen YW, Hossain MS, Chen PC, Tzeng SF, Chen PS, Kuo YM (2022) High-fat diet induces depression-like phenotype via astrocyte-mediated hyperactivation of ventral hippocampal glutamatergic afferents to the nucleus accumbens. Mol Psychiatry 27:4372–4384.
Uehara T, Sumiyoshi T, Itoh H, Kurachi M (2007) Role of glutamate transporters in the modulation of stress-induced lactate metabolism in the rat brain. Psychopharmacology (Berl) 195:297–302.
Wang L, Hu X, Ren Y, Lv J, Zhao S, Guo L, Liu T, Han J (2023) Arousal modulates the amygdala-insula reciprocal connectivity during naturalistic emotional movie watching. Neuroimage 279:120316.
Wood OWG, Yeung JHY, Faull RLM, Kwakowsky A (2022) EAAT2 as a therapeutic research target in Alzheimer’s disease: A systematic review. Front Neurosci 16:952096.
Xiao Q, Xu X, Tu J (2020) Chronic optogenetic manipulation of basolateral amygdala astrocytes rescues stress-induced anxiety. Biochem Biophys Res Commun 533:657–664.
Zhang X, Asim M, Fang W, Md Monir H, Wang H, Kim K, Feng H, Wang S, Gao Q, Lai Y, He J (2023) Cholecystokinin B receptor antagonists for the treatment of depression via blocking long-term potentiation in the basolateral amygdala. Mol Psychiatry 28:3459–3474.

Date Created: 20240806 Latest Revision: 20240808

20240808

10.4103/NRR.NRR-D-23-01411

39104111