Sodium diethyldithiocarbamate, FeSO4 7H2O, diethylenetriaminepentaacetic acid, and glutathione ethyl ester were purchased from Sigma‐Aldrich. N‐Hydroxysulfosuccinimidobiotin was obtained from Merck, dithiothreitol from Promega, streptavidin–Sepharose from GE Healthcare Bio‐Sciences, and Protein A/G‐Plus agarose from Santa Cruz Biotechnology. Osmotic mini‐pumps were purchased from Alzet and CL from Sigma‐Aldrich. Dihydroethidium and N‐(Biotinoyl)‐N′‐(iodoacetyl) ethylenediamine were obtained from Invitrogen. Monoclonal antibodies were purchased from the following vendors: β1 subunit of Na+‐K+ ATPase from Upstate Biotechnology; anti‐glutathione from Virogen, eNOS antibody from Sigma‐Aldrich, and p47phox, phosphorylated eNOS116, phosphorylated eNOS1177, neuronal NOS (nNOS), and α tubulin from Santa Cruz Biotechnology. All chemicals used in Krebs solutions were analytical grade and were obtained from BDH.
Osmotic minipumps
Manufactured by Merck Group
Sourced in United States
Osmotic minipumps are a type of laboratory equipment used for the continuous and controlled delivery of substances over an extended period. They operate based on the principle of osmosis, where the gradual influx of water into the device's reservoir causes the release of the contained substance at a predetermined rate.
Automatically generated - may contain errors
Lab products found in correlation
Isoproterenol, by Merck Group (2 mentions)
Sodium diethyldithiocarbamate, by Merck Group (1 mentions)
Dihydroethidium, by Thermo Fisher Scientific (1 mentions)
Glutathione ethyl ester, by Merck Group (1 mentions)
Dithiothreitol (dtt), by Promega (1 mentions)
P47phox, by Merck Group (1 mentions)
Diethylenetriaminepentaacetic acid, by Merck Group (1 mentions)
Streptavidin sepharose, by GE Healthcare (1 mentions)
Feso4 7h2o, by Merck Group (1 mentions)
Protein a g plus agarose, by Santa Cruz Biotechnology (1 mentions)
α tubulin, by Santa Cruz Biotechnology (1 mentions)
Bp 2010a, by Softron (1 mentions)
Isoflurane, by Baxter (1 mentions)
Aldosterone, by Merck Group (1 mentions)
5 protocols using osmotic minipumps
1
Biochemical Assay Reagents and Antibodies
2
NAMPT Haplodeficient Mice Model for Angiotensin II-Induced Hypertension
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Animal studies were approved by the Animal Care and Use Committee of Tongji Hospital Affiliated to Tongji University. The NAMPT haplodeficient mice (NAMPT+/-) were generated using CRISPR-Cas9 technology, as described previously [18 (link)]. The disruption of NAMPT by homologous recombination results in early embryonic lethality because of its dysfunction in egg cylinder organization and gastrulation. At the age of 8 weeks, male NAMPT+/- mice and their littermate wild-type (WT) mice were subcutaneously implanted with osmotic minipumps (model 2004; Alzet) to permit an infusion of Ang II (Sigma-Aldrich, Cat. no. A9525, 0.7 mg/kg per day) or saline for 4 weeks under inhalation anesthesia with 2% v/v isoflurane/oxygen, as described previously [44 (link)]. Blood pressure was measured weekly with a tail-cuff system using a heated scanner unit (BP-2010A, Softron) after surgery. The blood pressure of each mouse was detected at least five times until reaching a steady state, and the final value was calculated from an average of three proximal repeats.
3
Assessing Cardiac Function in Mice
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Osmotic minipumps (Alzet, Cupertino, CA, USA) were implanted subcutaneously in WT or Nur77-KO mice. The procedure was performed under isoflurane anaesthesia (4% isoflurane for induction, 2.5% isoflurane and O2 for maintenance of anaesthesia; Baxter, Illinois, United States) with subcutaneously-administered Carprofen (4 mg/kg) as a local analgetic. The Osmotic minipumps continuously released isoproterenol (Sigma) dissolved in saline, in a dose of 60 mg/kg/day. Control mice received minipumps containing saline only. After 7 days the mice were euthanized through a lethal dose of intraperitoneal-injected ketamine (238 mg/kg)/xylazine (102 mg/kg). Prior to and 6 days after implantation of the Osmotic minipumps, transthoracic echocardiography was performed in isoflurane-anaesthetized mice (4% isoflurane for induction, 2.5% isoflurane and O2 for maintenance of anaesthesia). Using the Vevo770 imaging system equipped with a 30-MHz linear array transducer (VisualSonics Inc, Toronto, Canada), a two-dimensional short-axis view of the left ventricle was obtained at the level of the papillary muscles and 2D M-mode tracings were recorded. Left ventricular ejection fraction and fractional shortening were calculated using the following formulas: LVEF% = (LVvol;d-LVvol;s)/(LVvol;d)*100; LVFS% = (LVID;d-LVID;s)/(LVID;d)*100.
4
Isoproterenol-Induced Cardiac Hypertrophy
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WT, Nur77-KO, cardiomyocyte-specific Nur77-deficient mice (CM-KO) and their control littermates (CM-WT) were employed in previous studies [21 (link),25 (link)]. Briefly, osmotic minipumps (Alzet, Cupertino, CA, USA) were implanted subcutaneously under isoflurane anesthesia (4% isoflurane for induction, 2.5% isoflurane and O2 for maintenance of anesthesia; Baxter, IL, United States) with subcutaneously administered carprofen (4 mg/kg) as a local analgesic. osmotic minipumps contained 60 mg/kg/day isoproterenol (Sigma, St. Louis, MO, USA) or saline as control. After 7 days, the mice were euthanized through a lethal dose of intraperitoneal-injected ketamine (238 mg/kg)/xylazine (102 mg/kg), after which hearts were excised.
5
Aldosterone-Induced Hypertension Model in Mice
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We used and defined an aldosterone‐induced hypertension model that has been previously reported by Brilla et al
29 (link) as an MR‐related hypertension model in this study. Eight‐week‐old male cardiac‐specific and inducible KO (c‐MYPT2−/−) mice and their littermate MYPT2f/f (MYPT2+/+) mice were divided into the following groups (n=10 each): (c‐MYPT2−/− control), MYPT2+/+ control, c‐MYPT2−/− MR‐related hypertension (c‐MYPT2−/−‐Treated), and MYPT2+/+ MR‐related hypertension (MYPT2+/+‐Treated). MR‐related hypertension groups were uni‐nephrectomized. After that, an osmotic minipump (model 1002; Alzet, Cupertino, CA) was implanted subcutaneously to infuse vehicle or aldosterone (0.15 μg/h; Sigma‐Aldrich, St. Louis, MO) in control or treated groups for 4 weeks, as previously described.
30 (link),
31 (link) Osmotic minipumps were replaced every 2 weeks. Heat support was provided throughout the procedure and during the recovery period. MR‐related hypertension mice were fed a high‐salt diet (8% NaCl), and control mice were fed a normal diet.
29 (link) as an MR‐related hypertension model in this study. Eight‐week‐old male cardiac‐specific and inducible KO (c‐MYPT2−/−) mice and their littermate MYPT2f/f (MYPT2+/+) mice were divided into the following groups (n=10 each): (c‐MYPT2−/− control), MYPT2+/+ control, c‐MYPT2−/− MR‐related hypertension (c‐MYPT2−/−‐Treated), and MYPT2+/+ MR‐related hypertension (MYPT2+/+‐Treated). MR‐related hypertension groups were uni‐nephrectomized. After that, an osmotic minipump (model 1002; Alzet, Cupertino, CA) was implanted subcutaneously to infuse vehicle or aldosterone (0.15 μg/h; Sigma‐Aldrich, St. Louis, MO) in control or treated groups for 4 weeks, as previously described.
30 (link),
31 (link) Osmotic minipumps were replaced every 2 weeks. Heat support was provided throughout the procedure and during the recovery period. MR‐related hypertension mice were fed a high‐salt diet (8% NaCl), and control mice were fed a normal diet.
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