The de novo Autism Spectrum Disorder RELN R2290C Mutation Reduces Reelin Secretion and Increases Protein Disulfide Isomerase Expression. Mus musculus

To determine if changes in Protein Disulfide Isomerase (PDIA1) expression in mice with a null Reelin allele were caused by accumulation of intracellular Reelin or were an effect of reduced Reelin protein, we examined expression of PDIA1 and other stress markers in heterozygous RELN +/- null allele mice. The levels of PDIA1 as well as PERK, BIP, phospho-eIF2alpha and total eIF2alpha were unchanged between wild-type and RELN +/- null allele mice. This suggested that there are phenotypic differences in the cerebella between mice that carry a RELN allele that fails to produce a protein (null allele) and those that make a protein that fails to be secreted (Orl allele). Each of the three major ER stress pathways ultimately leads to changes in gene transcription. Thus, we compared wild-type and heterozygous RELN Orl +/- mice cerebellum by RNAseq. Analysis was performed on 3 heterozygous (HET) and 3 wild-type (WT) cerebella, obtained from 6-week old male mice. Overall design: Comaprison of gene expression in the cerebellum of Reelin heterozygous null mice and wildtype mice using RNA-seq

为明确携带瑞林基因（RELN）无效等位基因的小鼠体内蛋白质二硫键异构酶（Protein Disulfide Isomerase, PDIA1）的表达变化，究竟是由细胞内瑞林（Reelin）蛋白积累所致，还是瑞林蛋白水平降低引发的效应，我们对杂合型RELN +/-无效等位基因小鼠的PDIA1及其他应激标志物的表达情况进行了检测。野生型与RELN +/-无效等位基因小鼠的PDIA1水平，以及PERK、BIP、磷酸化eIF2α与总eIF2α水平均未发生显著变化。这提示，携带无法合成蛋白的RELN等位基因（无效等位基因）的小鼠，与合成蛋白但无法分泌的Orl等位基因小鼠，二者的小脑表型存在差异。三大内质网应激通路最终均会介导基因转录改变，因此我们通过RNA测序（RNA-seq）比较了野生型与杂合型RELN Orl +/-小鼠的小脑组织基因表达谱。本研究共采集6周龄雄性小鼠的3份杂合型（HET）与3份野生型（WT）小脑组织开展分析。整体实验设计：采用RNA-seq技术比较瑞林杂合无效等位基因小鼠与野生型小鼠小脑的基因表达差异。