GluN2A-mediated currents and calcium signal in human iPSC-derived neurons
Gene expression data indicate that during human brain development, neurons change the NMDA receptor (NMDAR) subunit composition to modulate their function, favouring the GluN2A subunit over GluN2B-a hallmark of neuronal maturation. However, evidence supporting this phenomenon in human iPSC-derived neurons remains elusive. Here, using two differentiation methods in parallel (BrainPhys Neuronal Medium, BPM, and Neural Maintenance Medium, NMM), we provide evidence of increased synaptic localization of NMDARs during neuronal maturation and that GluN2A subunit is crucial for the NMDA physiological function-inducing inward currents and calcium entrance at 60 days of differentiation. Calcium responses to specific agonists, particularly NMDA, were elevated in cells cultured under BPM conditions. This is likely attributable to their more mature neuronal phenotype and the RNA-seq identified upregulation of genes involved in intracellular calcium signaling proteins. Our results offer insight into how glutamate receptor subunits mature during brain development, delineating approaches to study NMDAR activity in health and disease.
Authors
- Ana V. Paternain (ORCID: https://orcid.org/0000-0002-7128-4472)
- Federico Salas-Lucia (ORCID: https://orcid.org/0000-0003-4141-5790)
- Sergio Escamilla (ORCID: https://orcid.org/0000-0003-0952-7506)
- Elvira de la Peña
- Carlos Avilés-Granados (ORCID: https://orcid.org/0000-0001-5966-6737)
- Inmaculada Cuchillo-Ibáñez
- Francisco Andrés Peralta
- Javier Sáez-Valero
- Henrik Zetterberg
- María-Ángeles Cortés-Gómez
Institutions
- University of Alicante (ES)
- University of Wisconsin–Madison (US)
- Sahlgrenska University Hospital (SE)
- University of Chicago (US)
- Biomedical Research Networking Center on Neurodegenerative Diseases (ES)
- Instituto de Neurociencias (ES)
- Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (ES)
- UK Dementia Research Institute (GB)
- National Hospital for Neurology and Neurosurgery (GB)
- University College London (GB)
- Indian Institute of Science Bangalore (IN)
- University of Gothenburg (SE)
Publication Details
- Journal
- Scientific Reports
- Published
- 2026-02-18
- DOI
- https://doi.org/10.1038/s41598-026-38482-y
- Primary Topic
- Neuroscience and Neuropharmacology Research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Generalitat Valenciana
- Ministerio de Ciencia e Innovación
- Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas
- Instituto de Salud Carlos III
- European Regional Development Fund
- Agencia Estatal de Investigación