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创建媒介条件删除来自发育大脑皮尔金耶细胞的floxed等位基的工具

Jennifer N Jahncke¹, Kevin M Wright²

¹Neuroscience Graduate Program, Oregon Health & Science University, Portland, Oregon 97239.

|May 22, 2024

概括

在小鼠中评估Cre-lox系统以准小脑Purkinje细胞 (PCs) 显示,记者表达并不总是等于蛋白质损失. 一个Calb1 Cre线有效地准胚胎PC,而其他线则显示这些关键神经元的重组变化或不存在.

科学领域:

神经科学是一个神经科学.
遗传学是一个遗传学.
分子生物学分子生物学

背景情况:

在神经科学研究中,Cre-lox系统对于向基因操纵至关重要.
大脑皮尔金耶细胞 (PCs) 对于运动控制和学习至关重要.
精确准PC中的基因删除对于理解其功能至关重要.

研究的目的:

评估各种Cre-driver系的疗效以及针对小鼠小脑Purkinje细胞 (PCs) 的病毒方法.
评估Cre介导的重组与随后的蛋白质损失之间的相关性,特别是对于Dystroglycan (Dag1) 基因.
描述PC中不同Cre向策略的重组和蛋白质损失的发育时间.

主要方法:

使用了Cre-lox系统与光记者 (Ai14) 和化Dystroglycan (Dag1) 鼠标线.
在使用多个Cre驱动线 (Pcp2, Ptf1a, Nestin, Calb1) 的Purkinje细胞中评估了Cre介导的重组和Dag1蛋白损失.
在出生后的第0天 (P0) 采用Adeno相关病毒血清型8 (AAV8) 介导的Cre复合酶.

主要成果:

Pcp2 Cre线在PCs中显示了P7-P14的马赛克重组,Dag1蛋白质的损失直到P30才完全.
由于短暂的前体表达,Ptf1a Cre导致PC和内部神经元中随机的Dag1蛋白损失.
尼斯丁Cre没有驱动PC的重组,而Calb1Cre使胚胎PC的有效向和早期的Dag1损失成为可能;AAV8-Cre的输送导致了逐渐的PC转导和延迟的Dag1损失 (由P35).

结论:

不同的Cre-driver线和病毒方法在准小脑PC时表现出不同的效率和时间概况.
报告者基因表达并不总是准确地反映功能性蛋白质淘汰,突出显示了蛋白质水平验证的必要性.
Calb1 Cre系列是有效的向胚胎PCs,使基因删除在synaptogenesis之前,而其他工具提供不同的发育窗口.

关键词:

在 AAV AAV AAV 中.普尔金耶细胞是什么?小脑小脑是什么意思有条件的淘汰赛.在这里,我们将创建CRE.消极葡萄糖可以是消极葡萄糖

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