Effect of androgen; gonad- and androgen receptor on gene expression in mouse kidney and liver

To investigate mechanisms underlying renal sex differences, we performed whole kidney RNA-seq on wild type C57BL/6J mice (Jackson Laboratory) at postnatal 0, 2, 4, 8 and 79 weeks old. We also profiled wild type adult C57BL/6NCrl (Charles River Laboratory) and adult castrated, ovariectomized mice when the surgeries for gonadal removal were performed at the age of around 3 weeks old. Tissue specific Ar (androgen receptor), Tbx10 and Neurog2 knockout mice were maintained at F1 cross Arc/c floxed allele to Six2TGC/+ CRE strain. To examine the acute response of testosterone, we injected testosterone to adult castrated and ovariectomized mice and collected kidneys 24 hours later for bulk RNA-seq. To compare the mechanisms of Ar in regulating sex differences systemetically or locally in kidney and liver, we obtained bulk RNA-seq for kidneys and livers from ubiquitous, nephron-specific and hepatocyte-specific knockout of Ar by Sox2CRE/+, Six2TGC/+ and Albumin-CRE mouse strain respectively. Total RNA were collected for RNA-seq from minced 1) kidneys of C57BL/6J (Jackson Laboratory) at postnatal age 0, 2, 4, 8 and 79 weeks old; 2) kidneys of wild type C57BL/6NCrl mice (Charles River Laboratory), and those underwent castration or ovariectomy at around 3 weeks old that were injected with testosterone or oil control at 8~11 weeks old; 3) kidneys of adult conditional knockout by nephron-specific CRE strain Six2TGC/+ for Ar, Tbx10 and Neurog2 ; 4) kidneys and livers from ubiquitous removal of Ar by Sox2-CRE strain; 5) livers of hepatocyte-specific removal of Ar by Albumin CRE for both sexes.为探究肾脏性别差异的潜在分子机制，本研究对野生型C57BL/6J小鼠（杰克逊实验室（Jackson Laboratory））在出生后0、2、4、8及79周龄时开展全肾脏RNA测序（RNA-seq）。 我们同时对野生型成年C57BL/6NCrl小鼠（查尔斯河实验室（Charles River Laboratory）），以及于约3周龄接受性腺摘除手术的成年去势、卵巢切除小鼠进行了转录组分析。 组织特异性雄激素受体（Ar, androgen receptor）、Tbx10及Neurog2基因敲除小鼠通过将Arc/c floxed等位基因与Six2TGC/+ CRE品系进行F1杂交培育获得。 为检测睾酮的急性应答效应，我们向成年去势及卵巢切除小鼠注射睾酮，并于24小时后采集肾脏组织进行批量RNA测序（bulk RNA-seq）。 为对比雄激素受体（Ar）在肾脏与肝脏内系统性或局部调控性别差异的分子机制，我们分别通过Sox2CRE/+、Six2TGC/+及Albumin-CRE小鼠品系，构建了全身、肾单位特异性及肝细胞特异性的Ar敲除模型，并采集其肾脏与肝脏组织进行批量RNA测序。 本研究共收集以下五类样本用于RNA测序：1）出生后0、2、4、8及79周龄的C57BL/6J小鼠（杰克逊实验室）肾脏组织；2）野生型C57BL/6NCrl小鼠（查尔斯河实验室）肾脏组织，以及在约3周龄接受去势或卵巢切除手术、并于8~11周龄注射睾酮或油溶剂对照的小鼠肾脏组织；3）通过肾单位特异性CRE品系Six2TGC/+构建的成年Ar、Tbx10及Neurog2条件性敲除小鼠肾脏组织；4）通过Sox2-CRE品系实现全身Ar敲除小鼠的肾脏与肝脏组织；5）雌雄两性肝细胞特异性Ar敲除（由Albumin CRE构建）小鼠的肝脏组织。