Fos binding in hippocampal chromain 1 h after peritoneal KA administration

Fos induction is the most commonly used marker of neuronal activation. Here, we investigate the binding profile of Fos in the adult mouse hippocampus using antibodies specific against this transcription factor and the chromatin Immunoprecipitation coupled to DNA-sequencing (ChIP-seq) method. We generated profiles in basal conditions and 1 hour after neuronal stimulation. To trigger the syncronous activation of hippocampal exciatory neurons we treated the mice with kainic acid (KA) through peritoneal administration. This model of status epilepticus has been characterized in detail at the epigenomic level (Fernandez-Albert et al., 2019). Overall design: Two-month-old C57BL/6J mice were treated with PBS (saline) of Kainic acid (KA, two consecutive intraperitoneal injections separated by 30 min at 10 mg/kg). One hour after the first kainate injection, mice with level 4 epileptic seizures were sacrificed with subsequent dissection of their hippocampi. A Dounce homogenizer was used to dissociate hippocampal tissue and the extract was incubated with anti-Fos antibody and used for ChIP-seq.

Fos诱导（Fos induction）是目前最常用的神经元激活标记物。本研究利用针对该转录因子的特异性抗体，结合染色质免疫共沉淀联用DNA测序（chromatin Immunoprecipitation coupled to DNA-sequencing，ChIP-seq）技术，探究成年小鼠海马体中Fos的结合谱。我们分别在基础状态下以及神经元刺激后1小时获取了结合谱数据。为同步激活海马兴奋性神经元，我们通过腹腔注射的方式向小鼠施加红藻氨酸（kainic acid，KA）。该癫痫持续状态模型已在表观基因组层面得到了详尽的特征解析（Fernandez-Albert等，2019）。实验整体设计：将2月龄C57BL/6J小鼠分为两组，分别给予磷酸盐缓冲液（PBS，生理盐水）或红藻氨酸（KA：按10 mg/kg剂量连续两次腹腔注射，两次注射间隔30分钟）。首次注射红藻氨酸1小时后，对出现4级癫痫发作的小鼠实施安乐死，并随后分离其海马组织。使用杜恩匀浆器（Dounce homogenizer）解离海马组织，将组织提取物与抗Fos抗体孵育后，用于ChIP-seq实验。