Inhibition of TRPV4 rescues impaired excitability and social deficits in NAc-specific Shank3-deficient mice.

We used AAV transfection of Shank3 shRNA to study the effect of Shank3 knock down in Nucleus Accumbens neurons. We find that Shank3 knock down leads to social deficits and hyperexcitability of Nucleus Accumbens D1-MSN. Bulk RNA sequencing shows that D1-MSN with Shank3 knock down upregulate TRPV4 and its subsequent antagonist driven inhibition leads to both a rescue of D1-MSN hyperexcitability and social deficit. Overall design: Analysis of Nucleus accumbens cells from experiments involving the controls AAV-Scrambled or the AAV-Shank3-Sh Virus isolating in each case 4 different types of samples: Drd1 expressing infected cells, Drd1 expressing non-infected cells, non-Drd1 expressing infected cells and non-Drd1 expressing non-infected cells.

本研究采用腺相关病毒（AAV）介导的Shank3短发夹RNA（shRNA）转染，探究伏隔核（Nucleus Accumbens）神经元中Shank3基因敲低的生物学效应。研究发现，Shank3基因敲低会引发伏隔核D1型中型多棘神经元（D1-MSN）的社交行为缺陷与过度兴奋性升高。批量RNA测序结果显示，Shank3敲低的D1-MSN中瞬时受体电位阳离子通道蛋白V亚家族成员4（TRPV4）表达上调；后续通过TRPV4拮抗剂介导的抑制，可同时挽救D1-MSN的过度兴奋性表型与社交缺陷。整体实验设计：以AAV-乱序对照病毒（AAV-Scrambled）或AAV-Shank3-sh病毒感染的小鼠伏隔核细胞为研究对象，每组分别分离四类样本：表达多巴胺D1受体（Drd1）的感染细胞、表达多巴胺D1受体（Drd1）的未感染细胞、不表达多巴胺D1受体（Drd1）的感染细胞，以及不表达多巴胺D1受体（Drd1）的未感染细胞，并对上述样本进行分析。