RNA-Seq of Sprague Dawley Rat Skin Treated with Aldosterone or Vehicle on High-Salt Diet for 2 or 14 Days
RNA-Seq of Sprague Dawley Rat Skin Treated with Aldosterone or Vehicle on High-Salt Diet for 2 or 14 Days
Rats were euthanized by cervical dislocation. Carcasses, skin, and plasma were collected from aldosterone- and vehicle-treated rats after 2 and 14 days on a high-salt diet for the measurement of Na⁺, K⁺, and Cl⁻ concentrations. Skin samples for RNA extraction were flash-frozen and homogenized using a Tissuelyser (Qiagen).  Rats weighing approximately 300 g underwent unilateral nephrectomy and were individually housed with free access to tap water and a low-salt diet (AIN-76A diet containing 0.26% NaCl and 0.36% potassium; Diet No. 100000, Dyets Inc., Bethlehem, PA). Four weeks post-surgery, rats were randomized to receive a continuous subcutaneous infusion of either aldosterone or vehicle using subcutaneously implanted osmotic minipumps (2 pumps per rat). Aldosterone was infused at a dose of 1.5 microgram/h with a flow rate of 0.3 microliter/h. Following minipump implantation, rats were maintained on the low-salt diet and tap water for two weeks to recover. Subsequently, all rats were switched to a high-salt diet (AIN-76A diet containing 4% NaCl; Diet No. 113756, Dyets Inc., Bethlehem, PA) with tap water provided ad libitum. The high-salt diet was compositionally identical to the low-salt diet except for its increased salt content. RNA was isolated using the RNeasy Fibrous Tissue Kit (Qiagen) with water elution and stored at -80°C until further processing.  Library preparation was done with KAPA mRNA HyperPrep Kit for Illumina. Library size distribution was evaluated on the Agilent 2100 Bio-analyzer using the High Sensitivity DNA Kit (Agilent).