Characterization of a Novel Mutation in SLC1A1 Associated with Schizophrenia.
Characterization of a Novel Mutation in SLC1A1 Associated with Schizophrenia.
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DOI：
10.1159/000433599
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发表时间：
2015-10
期刊：
影响因子：
--
通讯作者：
Middleton FA
中科院分区：
文献类型：
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作者：
Afshari P;Myles-Worsley M;Cohen OS;Tiobech J;Faraone SV;Byerley W;Middleton FA
我们最近在一个扩展的帕劳家系中描述了SLC1A1基因染色体9p24.2的半缺失及其与精神分裂症的共分离。这一发现代表了精神分裂症的几种病理生理模型的趋同点。本报告试图描述这种半缺失的生物学后果。双荧光素酶测定表明，部分缺失的等位基因（缺乏外显子1和原生启动子）可以使用外显子2上游的序列作为替代启动子驱动5'截断的SLC1A1的表达。然而，共聚焦显微镜和电生理记录显示，5'截短的SLC1A1缺乏正常的膜定位和谷氨酸转运能力。为了确定半缺失的下游后果，我们首先使用主题qRT-PCR阵列来比较缺失携带者（n=11）与非携带者（n=8）以及患有精神病的缺失携带者（n=5）与没有精神病的人（n=3）外周血白细胞RNA中84个GABA和谷氨酸基因的表达。然后，使用靶向RNA-Seq （TREx）来量化SLC1A1敲低或SLC1A1全长或5'截短过表达的HEK293细胞中375个与神经精神疾病相关的基因的表达。检测到与精神分裂症病理生理密切相关的几个基因（如SLC1A2、SLC1A3、SLC1A6、SLC7A11、GRIN2A、GRIA1、DLX1）的表达变化。
We recently described a hemi-deletion on chromosome 9p24.2 at the SLC1A1 gene lcous and its co-segregation with schizophrenia in an extended Palauan pedigree. This finding represents a point of convergence for several pathophysiological models of schizophrenia. The present report sought to characterize the biological consequences of this hemi-deletion. Dual luciferase assays demonstrated that the partially-deleted allele (lacking exon 1 and the native promoter) can drive expression of a 5'-truncated SLC1A1 using sequence upstream of exon 2 as a surrogate promoter. However, confocal microscopy and electrophysiological recordings demonstrate that the 5'-truncated SLC1A1 lacks normal membrane localization and glutamate transport ability. To identify downstream consequences of the hemi-deletion we first used a themed qRT-PCR array to compare expression of 84 GABA and glutamate genes in RNA from peripheral blood leukocytes in deletion carriers (n=11) vs. non-carriers (n=8) as well as deletion carriers with psychosis (n=5) vs. those without (n=3). Then, targeted RNA-Seq (TREx) was used to quantify expression of 375 genes associated with neuropsychiatric disorders in HEK293 cells subjected to either knockdown of SLC1A1 or overexpression of full-length or 5'-truncated SLC1A1. Expression changes of several genes strongly implicated in schizophrenia pathophysiology were detected (e.g., SLC1A2, SLC1A3, SLC1A6, SLC7A11, GRIN2A, GRIA1, DLX1).