Td7034

Dahl salt-sensitive S (SS/jr) rats were obtained from the colony maintained by Dr. Michael Garrett at the University of Mississippi Medical Center. Female rats who had experienced two pregnancies at 12 and 17 weeks of age and virgin littermate controls were studied at six months of age, following a seven-week recovery from the second pregnancy. All pregnant animals displayed the previously published phenotype of superimposed preeclampsia [31 (link)]. These groups will hereafter be referred to as “virgin” (n=8) and “prior pregnancy” (n=11). Pups were removed within 48 hours of birth to remove any confounding effects of lactation. All rats were fed normal chow (TD7034, 0.3% NaCl, Harlan Teklad, Madison, WI) and water ad libitum on a 12-hour light/dark cycle. All experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were monitored by the University of Mississippi Medical Center Institutional Animal Care and Use Committee.

All animal procedures and protocols performed in this study were approved by the University of Toledo Institutional Animal Care and Use Committee. The procedure for the generation of a CRISPR/Cas9 targeted knock-in rat model has been previously described [3 (link)]. The non-founder S rat is a control derived from generating the knock-in. The nonfounder and the Spp2 knock-in rat strains were from stocks maintained at the University of Toledo animal facility. nonfounder and knock-in were not litter mates but were concomitantly bred for experimental procedures. Rats were weaned at 28 to 30 days of age and fed a low-salt diet (0.3% NaCl, TD 7034, Harlan Teklad). High-salt diet (2% NaCl, TD 94217, Harlan Teklad) was started at 6 weeks of age for all experiments.

Dahl salt-sensitive S (SS/jr) rats were obtained from the colony maintained by Dr. Michael Garrett at the University of Mississippi Medical Center and were fed normal chow (TD7034, 0.3% NaCl, Harlan Teklad, Madison, WI). Experiments were performed in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were monitored by the University of Mississippi Medical Center Institutional Animal Care and Use Committee. Sildenafil citrate (BIOTANG, Inc.) was administered at 50 mg/kg/day to a subset of rats in a gel diet beginning on gestational day (GD) 10 through GD 20. Blood pressure was measured via telemetry^{9 (link)}, and uterine artery resistance was measured via Doppler ultrasound^{12 (link)}. On GD 20, rats were anesthetized for tissue collection, and renal histology and biochemical assays were performed as previously described^{12 (link),13 (link)}.
All data are presented as mean ± SE. Statistical analyses were performed by either Student’s t-test (between the treated and control groups) or repeated measures ANOVA (MAP and proteinuria data) using Sigma Plot 12 (Systat Software, Inc.). Means were considered significantly different if p<0.05.

All animals were from a colony maintained at the University of Toledo College of Medicine and Life Sciences. All animal procedures and protocols used were approved by the University of Toledo Institutional Animal Care and Use Committee (IACUC protocol approval numbers 108854, 104573, 108390). Experiments were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and Animal Research Reporting of In Vivo Experiments (ARRIVE) guidelines. Male inbred Dahl salt-resistant (R) and salt-sensitive (S) rat strains were from the University of Toledo College of Medicine and Life Sciences. All rats were weaned at 4 weeks of age and maintained on a 12-hour light cycle with water ad libitum and a low-salt diet (0.3% NaCl, Harlan Teklad diet TD 7034; Madison, WI) until 5 weeks of age. At this time, the R and S were divided into two groups high-salt diet (2% NaCl, Harlan Teklad diet; TD 94217; Madison, WI) or maintained on a low-salt diet for a further 5–6 weeks.