In some studies, mice were rendered hypertensive via administration of DOCA and subsequent ad libitum access to isotonic saline drink (0.15M NaCl) in addition to continued access to standard drinking water.37 (link), 38 (link) In these studies, DOCA pellets (50 mg, Innovative Research of America, Sarasota, FL) were implanted subcutaneously (sc) in the interscapular region. Immediately after pellet implantation, mice were provided free access to isotonic saline drink for the three-week duration of the study. Controls underwent sham pellet implantation surgery.
Doca pellet
Manufactured by Innovative Research
Sourced in United States
DOCA pellets are a laboratory product used for research purposes. They are composed of deoxycholic acid, a naturally occurring bile acid. DOCA pellets are used in various research applications, but their core function is to serve as a component in experimental procedures. No further details or interpretations about their intended use can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
L name, by Merck Group (2 mentions)
Tail cuff method, by Kent Scientific (2 mentions)
Ta11pa c10, by Data Sciences International (1 mentions)
Lithium heparinate, by Merck Group (1 mentions)
125i pe13f, by PerkinElmer (1 mentions)
Sodium heparinate, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Nω nitro l arginine methyl ester l name, by Merck Group (1 mentions)
Sprague dawley rat, by Charles River Laboratories (1 mentions)
4 0 silk suture, by Johnson & Johnson (1 mentions)
14 protocols using doca pellet
1
DOCA-Induced Hypertension in Mice
2
DOCA-salt Induced Hypertension in Mice
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Floxed control and CD renin KO mice were anesthetized with 2% isoflorane and the right kidney was removed using a dorsal incision. Mice were monitored closely in the post-operative period for signs of pain or distress and treated with Rimadyl wafers as needed. Mice were monitored twice daily for the first 48 hours and then daily thereafter. After 7 days of recovery, radio telemetry devices (TA11-PAC10, Data Sciences International, St. Paul, MN) were inserted with the catheter implanted in the carotid artery. Mice were housed in individual cages, monitored daily for signs of pain or distress and allowed to recover for 5 days before recording BP. No adverse events were noted in the post-operative period following either surgery. After 3 days of baseline BP measurement on normal sodium diet (0.25% Na+, Harlan Teklad #2920X, Indianapolis, IN), DOCA pellets (50 mg, 21 day release, Innovative Research of America, Sarasota, FL) were implanted subcutaneously under isoflorane anesthesia. Mice were maintained on a normal sodium pellet diet and provided 0.9% saline for drinking water as per published DOCA-salt protocol [17 (link)]. Mice were not handled during BP recording since even small stimuli can markedly affect BP in mice.
3
Porcine Model of Hypertension-Induced Cardiac Tissue
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The animal experiments from which the samples of the present study were taken were approved by the local Bioethics Committee of Vienna, Austria (BMWF‐66.010/0091‐II/3b/2013) and conformed to the guide for the care and use of laboratory animals, US National Institute of Health (NIH Publication No. 85–23, revised 1996). Details of the animal model and experimental procedure were published in Schwarzl et al. (2015 ) and Reiter et al. (2016 ). Briefly, 10 female landrace pigs were assigned to a control or DOCA group (n = 5 each). In the latter, arterial hypertension was evoked by subcutaneous implantation of DOCA pellets (100 mg kg−1, 90‐day release, Innovative Research of America, USA) in combination with a high‐salt, high‐sugar, high‐potassium diet. The experiments were terminated at 12 weeks after the onset of treatment. After several in vivo measurements under appropriate anesthesia (Reiter et al. 2016 ), a bolus of 100 mmol potassium was injected into the coronary arteries to induce cardiac arrest and death of the animals. Subendocardial, midmyocardial and subepicardial samples were taken from the left ventricular free wall (excluding the interventricular septum) and fixed and stored in 1.5% paraformaldehyde, 1.5% glutaraldehyde in 0.15 m Hepes buffer at 4 °C.
4
Stroke and Salt-Loading Protocol
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Animals were weaned at age six weeks and randomly assigned to experimental or control groups. Isoflurane was used for both induction (5%) and maintenance (1.5–2%) of anesthesia. Salt-loaded animals (‘SBN/y-DOCA’ (n = 4) and ‘SBH/y-DOCA’ (n = 19) groups) were subcutaneously implanted with 75 mg deoxycorticosterone-acetate (DOCA) pellets (Innovative Research of America) and provided with 1% NaCl and 0.001% Potassium Chloride in tap water. Sham control animals (‘SBN/y sham’ (n = 4) and ‘SBH/y sham’ (n = 12) groups) were anesthetized, a skin incision was made, and the incision was sewn closed. Control animals were provided with tap water with no added salt. Animals were followed daily for a period of two months. When severe symptoms of stroke or loss of >10% body weight/week occurred, or it appeared that death was imminent, animals were sacrificed. Sacrificed animals were counted in survival curves, and tissues were harvested and used for analyses.
5
Peptide Synthesis and Radioligand Binding
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K17F was synthesized by GL Biochem (Shangai, China). LIT01-196 (300 mg) was synthesized by the Laboratoire d’Innovation Thérapeutique (CNRS UMR7200, Illkirch, France), as previously described (Gerbier et al., 2017 (link)). [125I]-pE13F (monoiodinated on Lys8 with Bolton-Hunter reagent, 2,200 Ci/mmol) was purchased from PerkinElmer (Wellesley, MA, United States). DOCA pellets (50 mg, 21-days release) were obtained from Innovative Research of America (Sarasota, FL, United States). Sodium heparinate, lithium heparinate, EDTA and N(ω)-nitro-L-arginine methyl ester (L-NAME) were obtained from SIGMA-ALDRICH (Saint-Quentin Fallavier, France). Rabbit polyclonal antibodies directed against the apelin fragment K17F were produced in the laboratory, as previously described (Reaux et al., 2001 (link)).
6
Renal Injury in DOCA-salt Hypertension
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Eight weeks old uninephrectomized (UNX) Sprague Dawley rats were purchased from Charles River Laboratories. The rats were subsequently assigned to one of two groups: (i) UNX controls provided with normal drinking water (UNX) (n = 12); (ii) deoxycorticosterone acetate (DOCA) and sodium chloride (DOCA-salt) rats receiving 60-day time release DOCA pellets (50 mg; Innovative Research of America, Sarasota, FL) and 0.9% NaCl and 0.2% KCl in drinking water for 4 weeks (UNX + DOCA) (n = 24). Then DOCA-salt rats were randomly selected and implanted with osmotic minipumps delivering a) vehicle (N = 12) or b) 16 mg/kg/day Serelaxin (N = 12) for another 4 weeks. When the rats were 16 weeks old, blood pressure, glomerular filtration rate (GFR), urinary albumin excretion, and cardiac echo were measured. The animals were then anesthetized to collect blood, and harvest the heart and kidney tissue. The animal protocol of this study has been approved by the IACUC of the University of Colorado Denver and all experimental methods and procedures were carried out in accordance with the approved guidelines.
7
Unilateral Nephrectomy and DOCA-Induced Hypertension
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Abcc6−/− and WT mice were unilaterally nephrectomised under isoflurane inhalation anaesthesia (2.2–2.6% isoflurane in 100% O2). Hind-paw withdrawal, blink reflex and respiratory rate were monitored in these mice. A 1.5-cm incision was made through the skin and abdominal muscle caudal to the rib cage. The renal artery and vein were ligated with 4-0 silk sutures (Ethicon) and the left kidney was removed. Skin and muscle layers were closed separately with 4-0 silk sutures. In addition, a 1-cm incision was made in the back, at the base of the neck to implant DOCA pellets subcutaneously (Innovative Research of America, Sarasota, Florida, USA), to provide a dose of 1 mg/kg/day. Mice were then administered water containing 0.9% NaCl and 0.2% KCl. Upon recovery, they were monitored each day for 19 days to measure SBP, DBP and HR using tail-cuff plethysmography48 .
8
DOCA-Salt Induced Hypertension Model
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At 12 weeks of age, unilateral nephrectomy was performed under anesthesia (isoflurane 3.5% induction followed by 1.5% maintenance dose) in WT and KO mice (n = 8/group), and a pellet containing DOCA was implanted in each mouse subcutaneously (DOCA pellet, 75 mg, 60-day release, Innovative Research of America, Sarasota, Florida, USA). Subsequently, these mice were exposed to salt by adding 1% NaCl to drinking water (DOCA groups). Unilateral nephrectomy was also performed in an additional group of WT and KO mice (n = 10/group), but these animals received no DOCA pellet and were given normal drinking water (SHAM groups). Animals were treated for 8 weeks overall.
9
DOCA-salt Induced Hypertension in Mice
10
DOCA-salt Induced Hypertension in Mice
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The systolic, diastolic, mean arterial BP, and heart rate of the mice were measured for 5 consecutive days by the noninvasive tail-cuff method (Kent Scientific). The mice in DOCA-salt group had their left kidneys removed. The mice were implanted subcutaneously with a DOCA pellet (Innovative Research of America) and provided with drinking water containing 1.0% NaCl and 0.2% KCl. The mice in the sham group only had their left kidney removed and were given tap water. Mice in NG-nitro-L-arginine methyl ester hydrochloride (L-NAME)-treated groups were administered L-NAME (Sigma) in drinking water for 4 weeks after the surgery.
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