
Differential development and electrophysiological activity in cultured cortical neurons from the mouse and cynomolgus monkey
Brief intro:
- Author: Xue-Yan Zhang, Jun Li, Cai-Juan Li, Ying-Qi Lin, Chun-Hui Huang, Xiao Zheng, Xi-Chen Song, Zhu-Chi Tu, Xiao-Jiang Li, and Sen Yan
- Journal: Neural Regen Res
- Doi: https://www.doi.org/10.4103/1673-5374.313056
- Publication Date: 2021 Apr 23
Products/Services used in the paper
Quotation shows PackGene:To investigate the effects of mutant HTT on cortical neurons at the cellular level, we used the adeno-associated virus (AAV) vector to express N-terminal HTT (1–230 amino acids) containing either 23Q or 109Q and a dsRED fluorescence protein (RFP). The AAV vectors were packaged and amplified by PackGene Biotech (Guangzhou, China). The vector contained the cytomegalovirus (CMV) promoter to express transgenes in cultured cortical neurons.
Research Field:CNS
Targeted organ:primary cortical neurons
Animal or cell line strain:Wild-type C57BL/6 mice, cynomolgus monkey
Abstract
In vitro cultures of primary cortical neurons are widely used to investigate neuronal function. However, it has yet to be fully investigated whether there are significant differences in development and function between cultured rodent and primate cortical neurons, and whether these differences influence the utilization of cultured cortical neurons to model pathological conditions. Using in vitro culture techniques combined with immunofluorescence and electrophysiological methods, our study found that the development and maturation of primary cerebral cortical neurons from cynomolgus monkeys were slower than those from mice. We used a microelectrode array technique to compare the electrophysiological differences in cortical neurons, and found that primary cortical neurons from the mouse brain began to show electrical activity earlier than those from the cynomolgus monkey. Although cultured monkey cortical neurons developed slowly in vitro, they exhibited typical pathological features-revealed by immunofluorescent staining-when infected with adeno-associated viral vectors expressing mutant huntingtin (HTT), the Huntington's disease protein. A quantitative analysis of the cultured monkey cortical neurons also confirmed that mutant HTT significantly reduced the length of neurites. Therefore, compared with the primary cortical neurons of mice, cultured monkey cortical neurons have longer developmental and survival times and greater sustained physiological activity, such as electrophysiological activity. Our findings also suggest that primary cynomolgus monkey neurons cultured in vitro can simulate a cell model of human neurodegenerative disease, and may be useful for investigating time-dependent neuronal death as well as treatment via neuronal regeneration. All mouse experiments and protocols were approved by the Animal Care and Use Committee of Jinan University of China (IACUC Approval No. 20200512-04) on May 12, 2020. All monkey experiments were approved by the IACUC protocol (IACUC Approval No. LDACU 20190820-01) on August 23, 2019 for animal management and use.
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