Department of Biomedical Sciences, Grand Valley State University, 1 Campus Drive, Allendale MI 49401, USA; Department of Cell and Molecular Biology, Grand Valley State University, 1 Campus Drive, Allendale MI 49401, USA.
Department of Cell and Molecular Biology, Grand Valley State University, 1 Campus Drive, Allendale MI 49401, USA.
Neuroscience. 2019 Nov 21;421:176-191. doi: 10.1016/j.neuroscience.2019.09.003. Epub 2019 Oct 28.
The floor plate of the developing midbrain gives rise to dopaminergic (DA) neurons, an important class of cells involved in Parkinson's disease (PD). Neural progenitors of the midbrain floor plate utilize key genes in transcriptional networks to drive dopamine neurogenesis. Identifying factors that promote dopaminergic neuron transcriptional networks can provide insight into strategies for therapies in PD. Using the chick embryo, we developed a quantitative PCR (qPCR) based method to assess the potential of a candidate factor to drive DA neuron gene expression, including the basic helix-loop-helix transcription factor Nato3 (Ferd3l). We then showed that overexpression of Nato3 in the developing chick mesencephalon produces a regionally dependent increase in genes associated with the DA neurogenesis, (such as Foxa2, Lmx1b and Shh) as well as DA neuron genes Nurr1 (an immature DA neuron marker) and mRNA expression of tyrosine hydroxylase (TH, a mature DA neuron marker). Interestingly, our data also showed that Nato3 is a potent regulator of Lmx1b by its broad induction of Lmx1b expression in neural progenitors of multiple regions of the CNS, including the midbrain and spinal cord. These data introduce a new, in vivo approach to identifying a gene that can drive DA transcriptional networks and provide the new insight that Nato3 can drive expression of key DA neuron genes, including Lmx1b, in neural progenitors.
中脑发育中的基板产生多巴胺能（DA）神经元，这是一类与帕金森病（PD）相关的重要细胞。中脑基板的神经前体细胞利用转录网络中的关键基因来驱动多巴胺神经发生。确定促进多巴胺能神经元转录网络的因素可以为 PD 的治疗策略提供深入了解。我们使用鸡胚开发了一种基于定量 PCR（qPCR）的方法来评估候选因子驱动 DA 神经元基因表达的潜力，包括基本螺旋-环-螺旋转录因子 Nato3（Ferd3l）。然后，我们表明，在发育中的鸡中脑中转染过量的 Nato3 会导致与 DA 神经发生相关的基因（如 Foxa2、Lmx1b 和 Shh）以及 DA 神经元基因 Nurr1（不成熟的 DA 神经元标记物）和酪氨酸羟化酶（TH，成熟的 DA 神经元标记物）的区域依赖性增加。有趣的是，我们的数据还表明，Nato3 通过广泛诱导中枢神经系统多个区域（包括中脑和脊髓）的神经前体细胞中 Lmx1b 的表达，成为 Lmx1b 的有效调节因子。这些数据引入了一种新的、体内方法来识别可以驱动 DA 转录网络的基因，并提供了新的见解，即 Nato3 可以驱动包括 Lmx1b 在内的关键 DA 神经元基因在神经前体细胞中的表达。
