Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan.
Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako City, Saitama 351-0198, Japan.
Cells. 2020 Dec 22;10(1):6. doi: 10.3390/cells10010006.
The common marmoset () has attracted considerable attention, especially in the biomedical science and neuroscience research fields, because of its potential to recapitulate the complex and multidimensional phenotypes of human diseases, and several neurodegenerative transgenic models have been reported. However, there remain several issues as (i) it takes years to generate late-onset disease models, and (ii) the onset age and severity of phenotypes can vary among individuals due to differences in genetic background. In the present study, we established an efficient and rapid direct neuronal induction method (induced neurons; iNs) from embryonic and adult marmoset fibroblasts to investigate cellular-level phenotypes in the marmoset brain in vitro. We overexpressed reprogramming effectors, i.e., microRNA-9/9*, microRNA-124, and Achaete-Scute family bHLH transcription factor 1, in fibroblasts with a small molecule cocktail that facilitates neuronal induction. The resultant iNs from embryonic and adult marmoset fibroblasts showed neuronal characteristics within two weeks, including neuron-specific gene expression and spontaneous neuronal activity. As directly reprogrammed neurons have been shown to model neurodegenerative disorders, the neuronal reprogramming of marmoset fibroblasts may offer new tools for investigating neurological phenotypes associated with disease progression in non-human primate neurological disease models.
普通狨猴（）因其能够重现人类疾病的复杂和多维表型而引起了相当大的关注，特别是在生物医学科学和神经科学研究领域，并且已经报道了几种神经退行性转基因模型。然而，仍然存在一些问题，即（i）生成迟发性疾病模型需要数年时间，并且（ii）由于遗传背景的差异，表型的发病年龄和严重程度在个体之间可能有所不同。在本研究中，我们建立了一种从胚胎和成年狨猴成纤维细胞中高效快速诱导直接神经元的方法（诱导神经元；iNs），以研究体外狨猴大脑中的细胞水平表型。我们用一种促进神经元诱导的小分子鸡尾酒在成纤维细胞中过表达重编程效应因子，即 microRNA-9/9*、microRNA-124 和 Achaete-Scute 家族 bHLH 转录因子 1。来自胚胎和成纤维细胞的 iNs 在两周内表现出神经元特征，包括神经元特异性基因表达和自发神经元活动。由于直接重编程神经元已被证明可模拟神经退行性疾病，因此狨猴成纤维细胞的神经元重编程可能为研究与非人类灵长类动物神经疾病模型中疾病进展相关的神经表型提供新工具。
