N-methyl-d-aspartate Receptor-mediated Preconditioning Mitigates Excitotoxicity in Human Induced Pluripotent Stem Cell-derived Brain Organoids. (21st February 2022)

• Record Type: Journal Article
• Authors: Bauersachs, Hanke Gwendolyn; Bengtson, C. Peter; Weiss, Ursula; Hellwig, Andrea; García-Vilela, Celia; Zaremba, Bastienne; Kaessmann, Henrik; Pruunsild, Priit; Bading, Hilmar
• Is Part Of: Neuroscience. Volume 484(2022)
• Page Start: 83
• Page End: 97
• Publication Date: 2022-02-21
• Subjects: AMPA α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid -- BB blocking buffer -- BSA bovine serum albumin -- CRE cAMP response element -- CREB CRE binding protein -- ECL enhanced chemiluminescence -- GABAA γ-aminobutyric acid type A -- iPSC induced pluripotent stem cell -- LDH lactate dehydrogenase -- NM neural medium -- NMDA N-methyl-d-aspartate -- NMDAR N-methyl-d-aspartate receptor -- MM maintenance medium -- PBST PBS plus Tween -- pCREB phosphorylated CREB -- SEM scanning electron microscopy -- sEPSC spontaneous excitatory postsynaptic current -- sIPSC spontaneous inhibitory postsynaptic current -- snRNA-seq single nuclei RNA sequencing -- sPSC spontaneous postsynaptic current -- TBP TATA-box binding protein -- TEM transmission electron microscopy; human neuron -- glutamate -- neurodegeneration -- synaptic activity -- pro-survival signaling -- neuroprotection; Neurochemistry -- Periodicals; Neurophysiology -- Periodicals; Neurology -- Periodicals; Neurochimie -- Périodiques; Neurophysiologie -- Périodiques; Neurochemistry; Neurophysiology; Electronic journals; Periodicals; Electronic journals; 612.8
• DOI: 10.1016/j.neuroscience.2021.12.026
• Languages: English
• ISSNs: 0306-4522

Abstract

Graphical abstract: Highlights: NMDA receptors mediate glutamate excitotoxicity in human iPSC-derived brain organoids. Brain organoid excitotoxicity involves structural disintegration and CREB shut-off. Preconditioning with NMDA boosts synaptic activity and cell survival in organoids. Abstract: Studies in rodent models of acute and chronic neurodegenerative disorders have uncovered that glutamate-induced excitotoxic cell death is mediated primarily by extrasynaptic N-methyl-d -aspartate receptors (NMDARs). Rodent neurons can also build up in an activity-dependent manner a protective shield against excitotoxicity. This form of acquired neuroprotection is induced by preconditioning with low doses of NMDA or by activation of synaptic NMDARs triggered by bursts of action potentials. Whether NMDARs in human neurons have similar dichotomous actions in cell death and survival is unknown. To investigate this, we established an induced pluripotent stem cell (iPSC)-derived forebrain organoid model for excitotoxic cell death and explored conditions of NMDAR activation that promote neuronal survival when applied prior to a toxic insult. We found that glutamate-induced excitotoxicity in human iPSC-derived neurons is mediated by NMDARs. Treatment of organoids with high concentrations of glutamate or NMDA caused the typical excitotoxicity pathology, comprising structural disintegration, neurite blebbing, shut-off of the transcription factor CRE binding protein (CREB), and cell death. In contrast, bath-applied low doses of NMDA elicited synaptic activity, a robust and sustained increase in CREB phosphorylation as well as function, and upregulation of immediate-early genes, including neuroprotective genes. Moreover, we found that conditions of enhanced synaptic activity increased survival of human iPSC-derived neurons if applied as pre-treatment before toxic NMDA application. These results revealed that both toxic and protective actions of NMDARs are preserved in human neurons. The experimental platform described in this study may prove useful for the validation of neuroprotective gene products and drugs in human neurons.