Robust Expression of Functional NMDA Receptors in Human Induced Pluripotent Stem Cell-Derived Neuronal Cultures Using an Accelerated Protocol.

Autor:	Ruden JB; Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, United States., Dixit M; Division of Geriatric Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, United States., Zepeda JC; Department of Pharmacology, Vanderbilt University, Nashville, TN, United States., Grueter BA; Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, United States.; Department of Pharmacology, Vanderbilt University, Nashville, TN, United States.; Department of Anesthesiology, Vanderbilt University Medical Center, Nashville, TN, United States.; Department of Psychiatry, Vanderbilt University Medical Center, Nashville, TN, United States.; Vanderbilt Center for Addiction Research, Vanderbilt University, Nashville, TN, United States.; Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, TN, United States., Dugan LL; Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, United States.; Division of Geriatric Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, United States.; VA Tennessee Valley Geriatric Research, Education, and Clinical Center (GRECC), Nashville, TN, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in molecular neuroscience [Front Mol Neurosci] 2021 Nov 26; Vol. 14, pp. 777049. Date of Electronic Publication: 2021 Nov 26 (Print Publication: 2021).
DOI:	10.3389/fnmol.2021.777049
Abstrakt:	N-methyl-D-aspartate (NMDA) receptors are critical for higher-order nervous system function, but in previously published protocols to convert human induced pluripotent stem cells (iPSCs) to mature neurons, functional NMDA receptors (NMDARs) are often either not reported or take an extended time to develop. Here, we describe a protocol to convert human iPSC-derived neural progenitor cells (NPCs) to mature neurons in only 37 days. We demonstrate that the mature neurons express functional NMDARs exhibiting ligand-activated calcium flux, and we document the presence of NMDAR-mediated electrically evoked postsynaptic current. In addition to being more rapid than previous procedures, our protocol is straightforward, does not produce organoids which are difficult to image, and does not involve co-culture with rodent astrocytes. This could enhance our ability to study primate/human-specific aspects of NMDAR function and signaling in health and disease. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2021 Ruden, Dixit, Zepeda, Grueter and Dugan.)
Databáze:	MEDLINE
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