Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
Nuffield Department of Clinical Neurosciences, Academic Unit of Neuropathology, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DU, UK; The Oxford Parkinson's Disease Centre, University of Oxford, Oxford OX1 3QX, UK.
Stem Cell Reports. 2017 Aug 8;9(2):587-599. doi: 10.1016/j.stemcr.2017.06.005. Epub 2017 Jul 6.
The H1 haplotype of the microtubule-associated protein tau (MAPT) locus is genetically associated with neurodegenerative diseases, including Parkinson's disease (PD), and affects gene expression and splicing. However, the functional impact on neurons of such expression differences has yet to be fully elucidated. Here, we employ extended maturation phases during differentiation of induced pluripotent stem cells (iPSCs) into mature dopaminergic neuronal cultures to obtain cultures expressing all six adult tau protein isoforms. After 6 months of maturation, levels of exon 3+ and exon 10+ transcripts approach those of adult brain. Mature dopaminergic neuronal cultures display haplotype differences in expression, with H1 expressing 22% higher levels of MAPT transcripts than H2 and H2 expressing 2-fold greater exon 3+ transcripts than H1. Furthermore, knocking down adult tau protein variants alters axonal transport velocities in mature iPSC-derived dopaminergic neuronal cultures. This work links haplotype-specific MAPT expression with a biologically functional outcome relevant for PD.
微管相关蛋白 tau（MAPT）基因座的 H1 单倍型与神经退行性疾病（包括帕金森病）具有遗传相关性，并影响基因表达和剪接。然而，这种表达差异对神经元的功能影响尚未完全阐明。在这里，我们在诱导多能干细胞（iPSC）分化为成熟多巴胺能神经元培养物的过程中采用了延长的成熟阶段，以获得表达所有六种成年 tau 蛋白同工型的培养物。在 6 个月的成熟后，外显子 3+和外显子 10+转录本的水平接近成年大脑的水平。成熟的多巴胺能神经元培养物在表达上表现出单倍型差异，H1 表达的 MAPT 转录本比 H2 高 22%，H2 表达的外显子 3+转录本比 H1 高 2 倍。此外，敲低成年 tau 蛋白变体改变了成熟 iPSC 衍生的多巴胺能神经元培养物中的轴突运输速度。这项工作将特定单倍型的 MAPT 表达与与帕金森病相关的生物学功能结果联系起来。
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