Journal
EUROPEAN JOURNAL OF PHARMACOLOGY
Volume 927, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.ejphar.2022.175056
Keywords
NMDA receptors; Primary neurons; iPSC; Neurosteroids
Categories
Funding
- Academy of Sciences of the Czech Republic [RVO 61388963]
- Technology Agency of the Czech Republic
- National Centres of Competence (NCK1) [TN01000013]
Ask authors/readers for more resources
The goal of this study was to evaluate the efficacy of mixed cortical and hippocampal primary rat postnatal neuronal culture for identifying NMDAR antagonists. The authors found that this model is comparable to other commonly used primary rat neuronal models and is a convenient and robust tool for NMDAR antagonist screening.
The goal of this study was to evaluate mixed cortical and hippocampal primary rat postnatal neuronal culture as in vitro tool for identification of N-methyl-D-aspartate receptor (NMDAR) antagonists and to find out, whether this model is comparable with other commonly used primary rat neuronal models differing in their origin (pure cortical vs. mixed cortical and hippocampal) and differentiation state (embryonal vs. postnatal). Induced pluripotent stem cell (iPSC) - derived human glutamatergic neurons have been included in this study as well. First, the cultures were characterized by their neuron/astrocyte composition, the mRNA expression of NR2B/ NR2A NMDAR subunit ratios, and the expression of glutamate transporters (GLT1, GLAST). Then, selected endogenous steroids and synthetic neuroactive steroids that have been previously identified as negative allosteric modulators of recombinant GluN1/GluN2B NMDA receptors, were evaluated for their ability to prevent an NMDA or glutamate-induced Ca2+ influx (acute effect) and excitotoxicity over 24 h. Though the neuroprotective potential against excitotoxic stimuli varied among the models studied, postnatal mixed cortical and hippocampal culture proved to be a convenient and robust tool for NMDAR antagonist screening. The most widely used embryonal (E18) cultures offered higher cell yields but at the expense of a higher sensitivity to compounds' cytotoxicity. iPSC-derived neurons were not found to be superior to rat cultures for screening purposes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available