Transcriptome profiling of embryonic cortex E18 and primary cortical neuron cultures deficient for the Amyloid Precursor Protein (APP)

The amyloid precursor protein (APP), an important player in the Alzheimer's disease, regulates neurites formation and synaptic function by mechanisms still not fully understood. Its ability to control gene expression via its intracellular domain (AICD) was proposed to mediate several of these physiological functions. Here, we tested whether putative AICD target genes could be transcriptionally modified by the absence of APP in embryonic cortex (E18) and in primary cortical neurons during maturation after 3 or 7 days in vitro. Moreover, we aimed to identified novel APP dependent target genes that could be related to particular biological functions in the context of neuronal physiology. Primary neuronal culture from embryonic cortices were carried out using APP+/+ and APP-/- embryos at developmental stage E18. Primary neuronal cultures were kept in vitro for 3 (DIV3) or 7 (DIV7) days and subjected to RNA extraction and hybridization on Affymetrix microarrays. In addition, embryonic cortices at E18 (E18) were subjected RNA extraction and hybrization on Affymetrix microarrays. In all our experimental conditions (E18, DIV3 and DIV7), 3 biologicals replicates for each genotype (APP+/+ and APP-/-) were used.淀粉样前体蛋白（amyloid precursor protein, APP）是阿尔茨海默病中的关键调控因子，其通过尚未完全阐明的机制调控神经突起形成与突触功能。有研究提出，其通过胞内结构域（intracellular domain, AICD）调控基因表达的能力，可介导上述多种生理功能的发挥。本研究分别在胚胎大脑皮层（E18）以及体外成熟培养3天（DIV3）或7天（DIV7）的原代皮层神经元中，探究了APP缺失是否会对推定的AICD靶基因产生转录水平的调控效应。此外，本研究旨在鉴定与神经元生理过程中特定生物学功能相关的新型APP依赖性靶基因。本研究选取发育阶段为E18的APP野生型（APP+/+）与APP敲除型（APP-/-）胚胎，分离其皮层组织制备原代神经元培养物。将原代神经元在体外分别培养3天（DIV3）与7天（DIV7）后，进行RNA提取并在Affymetrix基因芯片上完成杂交。此外，直接对E18时期的胚胎皮层组织进行RNA提取与Affymetrix基因芯片杂交。所有实验分组（E18、DIV3及DIV7）中，每种基因型（APP+/+与APP-/-）均设置3次生物学重复。