GSM5464002: Go.h; Homo sapiens; RNA-Seq
Serial kidney biopsies were performed in duplicate prior to pacing (Baseline), on Day-14 of pacing (HF), and after 25-days (non-pacing) recovery (6 sheep, 5 x Baseline, 6 x Heart Failure and 5 x Recovery serial samples). For the first two procedures, the sheep were anaesthetized (as described above) and 2 adjacent percutaneous kidney biopsies taken under ultrasonic guidance (Philips iE33; Philips Ultrasound, WA, USA) using a 14g spring-loaded automated biopsy gun (BARD Peripheral Vascular, Inc., Tempe, USA). Final kidney samples were collected at study-end immediately following euthanasia (IV Pentobarb300 100mg/kg). On each occasion, one of the biopsies was fixed in 10% formalin (4oC overnight) for histological analysis, and the other snap frozen in liquid nitrogen and stored at -80oC for subsequent RNA extraction and RNA-Seq analysis. To maximise statistical power, the 5 x Baseline, 6 x Heart Failure and 5 x Recovery serial samples were supplemented with kidney samples collected post-mortem from animals sacrificed before, during or after the same pacing protocol. This provided a total of 14 x Baseline, 13 x HF and 8 x Recovery samples for RNA-Seq analysis. Total RNA was extracted from frozen kidney biopsy samples and purified with RNeasy Midi Columns (Qiagen, Hilden, Germany).  RNA-Seq libraries were prepared using total RNA (500 ng input) and TruSeq stranded mRNA library prep kits (Illumina, San Diego, CA).