Single-molecule fluorescence in-situ hybridization reveals that human SHANK3 mRNA expression varies during development and in autism-associated SHANK3 heterozygosity.
Single-molecule fluorescence in-situ hybridization reveals that human SHANK3 mRNA expression varies during development and in autism-associated SHANK3 heterozygosity.
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单分子荧光原位杂交表明，在发育和自闭症相关的shank3杂合性中，人shank3 mRNA表达在变化。
DOI：
10.1186/s13287-018-0957-3
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发表时间：
2018-07-31
影响因子：
7.5
通讯作者：
Andreae LC
中科院分区：
文献类型：
--
作者：
Taylor SE;Taylor RD;Price J;Andreae LC
SHANK 3基因的缺失和突变与自闭症谱系障碍密切相关，并成为自闭症相关障碍M-M综合征的基础。SHANK 3是在兴奋性神经元的突触后膜处发现的支架蛋白。单分子荧光原位杂交（smFISH）允许在体外可视化单个mRNA转录物。在这里，我们在人诱导型多能干细胞（hiPSC）衍生的皮质神经元中进行并定量smFISH，靶向SHANK 3转录本。smFISH和常规免疫荧光染色均显示，在对照人类皮层神经元中，SHANK 3 mRNA和蛋白分别发育增加。对来自SHANK 3杂合型自闭症个体的神经元中的单个SHANK 3 mRNA分子的分析表明，虽然SHANK 3 mRNA转录物的数量在细胞索马中保持与对照水平相当，但在神经元过程中减少了50%，这表明SHANK 3 mRNA的局部树突靶向可能在SHANK 3单倍不足中受到特异性影响。人SHANK 3 mRNA在hiPSC衍生的神经元中显示发育调节的树突定位，其在从患有自闭症的单倍不足个体产生的神经元中减少。尽管需要进一步的复制，但考虑到局部mRNA翻译在突触功能中的重要性，这可能代表了重要的早期异常。
Deletions and mutations in the SHANK3 gene are strongly associated with autism spectrum disorder and underlie the autism-associated disorder Phelan–McDermid syndrome. SHANK3 is a scaffolding protein found at the post-synaptic membrane of excitatory neurons. Single-molecule fluorescence in-situ hybridization (smFISH) allows the visualization of single mRNA transcripts in vitro. Here we perform and quantify smFISH in human inducible pluripotent stem cell (hiPSC)-derived cortical neurons, targeting the SHANK3 transcript. Both smFISH and conventional immunofluorescence staining demonstrated a developmental increase in SHANK3 mRNA and protein, respectively, in control human cortical neurons. Analysis of single SHANK3 mRNA molecules in neurons derived from an autistic individual heterozygous for SHANK3 indicated that while the number of SHANK3 mRNA transcripts remained comparable with control levels in the cell soma, there was a 50% reduction within neuronal processes, suggesting that local, dendritic targeting of SHANK3 mRNA may be specifically affected in SHANK3 haploinsufficiency. Human SHANK3 mRNA shows developmentally regulated dendritic localization in hiPSC-derived neurons, which is reduced in neurons generated from a haploinsufficient individual with autism. Although further replication is needed, given the importance of local mRNA translation in synaptic function, this could represent an important early abnormality.
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影响因子：
30.8
作者：
Durand, Christelle M.;Betancur, Catalina;Bourgeron, Thomas
通讯作者：
Bourgeron, Thomas
影响因子：
5.2
作者：
Boccuto, Luigi;Lauri, Maria;Schwartz, Charles E.
通讯作者：
Schwartz, Charles E.
影响因子：
14.8
作者：
Kaech, Stefanie;Banker, Gary
通讯作者：
Banker, Gary
影响因子：
6.2
作者：
Betancur C;Buxbaum JD
通讯作者：
Buxbaum JD
患者特异性诱导的多能干细胞的效用用于自闭症谱系障碍的建模。
影响因子：
3.4
作者：
Cocks, Graham;Curran, Sarah;Gami, Priya;Uwanogho, Dafe;Jeffries, Aaron R.;Kathuria, Annie;Lucchesi, Walter;Wood, Victoria;Dixon, Rosemary;Ogilvie, Caroline;Steckler, Thomas;Price, Jack
通讯作者：
Price, Jack