Abstract:
Dopaminergic neurons in the brain are an important source of dopamine, which is a crucial neurotransmitter for wellbeing, memory, reward, and motor control. Deficiency of dopamine due to advanced age and accumulative dopaminergic neuron defects can lead to movement disorders such as Parkinson\'s disease. Glial cell-derived neurotrophic factor (GDNF) is one of many factors involved in dopaminergic neuron development and/or survival. However, other endogenous GDNF functions in the brain await further investigation. Zebrafish is a well-established genetic model for neurodevelopment and neurodegeneration studies. Importantly, zebrafish shares approximately 70% functional orthologs with human genes including GDNF. To gain a better understanding on the precise functional role of gdnf in dopaminergic neurons, our laboratory devised a targeted knockdown of gdnf in the zebrafish larval brain using vivo morpholino. Here, detailed protocols on the generation of gdnf morphants using vivo morpholino are outlined. This method can be applied for targeting of genes in the brain to determine specific spatiotemporal gene function in situ.
摘要:
大脑中的多巴胺能神经元是多巴胺的重要来源,这是幸福的关键神经递质,记忆,奖励,和电机控制。由于高龄和积累的多巴胺能神经元缺陷导致的多巴胺缺乏可导致运动障碍如帕金森病。胶质细胞源性神经营养因子(GDNF)是参与多巴胺能神经元发育和/或存活的许多因素之一。然而,脑内其他内源性GDNF功能有待进一步研究。斑马鱼是神经发育和神经变性研究的成熟遗传模型。重要的是,斑马鱼与包括GDNF在内的人类基因共享约70%的功能直向同源物。为了更好地了解gdnf在多巴胺能神经元中的确切功能作用,我们的实验室设计了一个有针对性的击倒gdnf在斑马鱼幼虫大脑中使用体内吗啉。这里,概述了使用体内吗啉代生成gdnf吗啡的详细方案。该方法可用于靶向大脑中的基因以原位确定特定的时空基因功能。