All animals underwent invasive hemodynamic testing on the euthanasia day. The rats were placed in the dorsal position on the operating table and anesthetized with pentobarbital sodium (30 mg/kg body weight, Biowet, Poland), which was administered intraperitoneally. Animals were mechanically ventilated during the whole procedure using a pressure-controlled respirator and a mixture of air and oxygen. Anesthesia was maintained by additional bolus doses of pentobarbital sodium as needed. Lidocaine (20 mg/mL, B. Braun Melsungen AG, Germany) was used for local infiltration of the surgical sites. The chest cavity was opened by the left and right mini-thoracotomy in the 6th intercostal space. For the measurement of ventricular systolic and end-diastolic pressure, the heparinized 21G venous cannulas connected to a pressure recording system (Siemens SC 7000, Erlangen, Germany) were introduced simultaneously to the right and left ventricles via their apexes [13 (link)]. The pressure transducer was fixed to the operating table and set at the level of the animal’s heart. The values were registered from a stable signal with 300-s periods, and mean values were calculated as output values. After the procedure, animals were euthanized via an overdose of sodium pentobarbital (Biowet, Poland) which was administered intraperitoneally.
Pentobarbital sodium
Manufactured by Biowet
Sourced in Poland, Germany
Pentobarbital sodium is a barbiturate that acts as a central nervous system depressant. It is commonly used as a sedative and hypnotic agent in laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Tween 80, by Merck Group (3 mentions)
Retinol, by Merck Group (2 mentions)
L glutathione reduced, by Merck Group (2 mentions)
L ascrobic acid, by Merck Group (2 mentions)
Chloro 2 4 dinitro benzene cdnb, by Merck Group (2 mentions)
5 5 dithiobis 2 dinitrobenzoic acid, by Merck Group (2 mentions)
Tx 100, by Merck Group (2 mentions)
Ketamine, by Biowet (2 mentions)
Xylazine, by Biowet (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
Sc 7 000, by Siemens (1 mentions)
Lidocaine, by B. Braun (1 mentions)
Butylated hydroxytoluene, by Merck Group (1 mentions)
C18 silica gel, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Acetic acid, by Chempur (1 mentions)
Ethanol, by Merck Group (1 mentions)
Sodium chloride (nacl), by Chempur (1 mentions)
Sr142948, by Bio-Techne (1 mentions)
1 fluoro 2 4 dinitrobenzene dnfb, by Merck Group (1 mentions)
11 protocols using pentobarbital sodium
1
Invasive Hemodynamic Assessment in Rats
2
Quantification of Oxidative Stress Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Drugs and reagents were obtained from the following sources: 3-(3-carbamoylphenyl)phenyl N-cyclohexylcarbamate (URB597), 8-iso Prostaglandin F2α-d4 (8-isoPGF2α–d4), 8-iso Prostaglandin F2α (8-isoPGF2α) anandamide-d8 (AEA-d8), and 2-arachidonylglycerol-d8 (2-AG-d8) from Cayman Chemical Company (Ann Arbor, MI, USA); dimethyl sulfoxide (DMSO), superoxide dismutase (SOD), catalase (CAT), L-ascrobic acid, retinol, L-glutathione reduced (GSH), and Tween 80 were acquired from Sigma-Aldrich (Steinheim, Germany); pentobarbital sodium was purchased from Biowet (Puławy, Poland), chloro-2,4-dinitro benzene (CDNB), butylated hydroxytoluene (BHT), and 5,5′-dithiobis (2-dinitrobenzoic acid) (DTNB) were acquired from Sigma-Aldrich (Steinheim, Germany). URB597 was dissolved in an URB597 solvent: a mixture of DMSO, Tween 80, and saline (0.9% NaCl) [1:2:7; v/v/v].
3
Extraction and Analysis of Organic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chemicals used in the experiment were as follows: trinitrobenzensulfonic acid (Sigma Aldrich, Germany); pentobarbital sodium (Biowet, Poland); physiological saline (Polpharma S.A., Poland); ethanol and formaldehyde (Avantor, Poland); acetic acid, methanol, butanol, diethyl ether, hexane (Chempur, Poland); acetonitrile (Merck, Germany) and C-18 silica gel (Sigma Aldrich, Germany).
4
Administering JZL195 and Pentobarbital in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Drugs were obtained from the following sources: (4-nitrophenyl) 4-[(3-phenoxyphenyl)methyl]piperazine-1-carboxylate (JZL195) from MedChemExpress (Monmouth Junction, NJ, USA); ethanol from POCH (Gliwice, Poland); Tween-80 from Sigma-Aldrich (Steinheim, Germany); pentobarbital sodium from Biowet (Puławy, Poland) and NaCl from Chempur (Piekary Śląskie, Poland). pentobarbital sodium was dissolved in saline and injected in a volume of 2 mL/kg. JZL195 was administered as a milky suspension at a volume of 1 mL/kg (at a dose of 1 and 10 mg/kg) and 2 mL/kg (at a dose of 100 mg/kg). The vehicle for JZL195 consisted of ethanol, Tween-80 and 0.9% saline mixed in a ratio of 3:1:16.
5
Neurotensin Receptor Antagonist Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neurotensin (NT) and neurotensin antagonist SR 142948 were purchased from Tocris Bioscience (Bristol, UK). 1-fluoro-2,4-dinitrobenzene (DNFB), dinitrobenzene sulfphonic acid (DNS), TX-100 and protease inhibitor cocktail were obtained from Sigma-Aldrich (Poznań, Poland). Pentobarbital sodium, ketamine and xylazine were acquired from Biowet (Puławy, Poland).
6
Oxidative Stress Biomarker Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Drugs and reagents were obtained from the following sources: 3-(3-carbamoylphenyl)phenyl N-cyclohexylcarbamate (URB597), 4-hyroxynonenal (4-HNE), 8-iso Prostaglandin F2α-d4 (8-isoPGF2α–d4), 8-iso Prostaglandin F2α (8-isoPGF2α) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) from Cayman Chemical Company (Ann Arbor, MI, USA); 11-deoxycorticosterone acetate (DOCA), dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), β-Nicotinamide adenine dinucleotide 2′-phosphate reduced tetrasodium salt hydrate (NADPH), xanthine, xanthine oxidase (XO), 9,9′-Bis(N-methylacridinium nitrate) (lucigenin), superoxide dismutase (SOD), catalase (CAT) thioredoxin (Trx), l -ascrobic acid, retinol, l -glutathione reduced (GSH), l -glutathione oxidized (GSSG), benzaldehyde-d6 and Tween 80 were acquired from Sigma-Aldrich (Steinheim, Germany); pentobarbital sodium was purchased from Biowet (Puławy, Poland); chloro-2,4-dinitro benzene (CDNB), butylated hydroxytoluene (BHT), and 5,5′-dithiobis (2-dinitrobenzoic acid) (DTNB) were acquired from Sigma-Aldrich (Steinheim, Germany). DOCA was dissolved in DMF, whereas URB597 was dissolved in an URB597 solvent: a mixture of DMSO, Tween 80, and saline (0.9% NaCl) [1:2:7; v/v/v].
7
Synthesis of Opioid Peptide PK20
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PK20 (H–Dmt–D–Lys–Phe–Phe–Lys–Lys–Pro–Phe–Tle–Leu–OH) was synthesized by using solid-phase peptide synthesis according to the method of Merrifield with Fmoc approach [77 (link)] in the Department of Neuropeptides. Chemicals and reagents were purchased from commercial companies: 1-fluoro-2, 4-dinitrobenzene (DNFB, No. 556971), dexamethasone (No. D4902), dinitrobenzene sulfonic acid (DNS, No. D1529), protease inhibitor cocktail (No. S8830), and TX-100 (No. X100) were obtained from Sigma-Aldrich (Poznań, Poland). Ketamine, xylazine, and pentobarbital sodium were purchased from Biowet (Puławy, Poland).
8
Bladder Reconstruction with BAM and ASCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
One hundred and four syngeneic male Wistar rats weighing between 250 and 300 g were randomly divided into eight equal groups (n = 13). Rats were anesthetized with sodium pentobarbital (15 mg/kg, i.p., Biowet, Poland) and lidocaine (20 mg/kg, i.m., Polfa, Poland) before undergoing hemicystectomy and bladder reconstruction with a graft of approximately 1 cm2 in size. Urinary bladders were reconstructed with BAM (first, second, third, and fourth groups) or BAM seeded with ASCs (fifth, sixth, seventh, and eighth groups). The animals were sacrificed after 7 (first and fifth groups), 30 (second and sixth groups), 90 (third and seventh groups), and 180 (fourth and eighth groups) days. The reconstructed bladders were harvested for macroscopic, histological, and molecular analyses.
9
Histological Verification of Microinjections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immediately after the completion of the experiments, rats were overdosed with sodium pentobarbital (morbital; 133.3 mg/ml; i.p.; Biowet, Poland) and the brains were removed and stored in a 4% formalin (POCH, Poland) solution for at least 3 days before the sectioning Brains were cut into 12-μm sections on a cryostat, mounted on gel-coated glass slides. The brain sections were defatted, stained with cresyl violet, cleared with xylene and placed under coverslips. The placement of microinjection probes were verified using a light microscope. There was no necrosis distal to the track upon histological examination of sections. Only data from rats with correctly placed probes within the NAc and the PFc according to previously established guidelines were included for statistical analyses (Fig. 2 ).![]()
Histological verification of microinjection representative probe placements in the NAc (left panels) and the PFc (right panels) of rats that underwent cocaine self-administration (
10
Neurotoxicity Evaluation of 25I-NBOMe
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
4-Iodo-2,5-dimethoxy-N-(2-methoxybenzyl)phenethylamine hydrochloride (25I-NBOMe) was purchased from Chiron AS (Trondheim, Norway). The chemicals used for the alkaline comet assay were from Trevigen (Gaithersburg, MD, USA) and Merck (Warszawa, Poland). TUNEL assay was performed with the usage of Click-iT™ Plus TUNEL Assay of Invitrogen, Thermo Fisher Scientific (Waltham, MA, USA). The reagents used in immunohistochemistry came from Sigma Aldrich (Poznań, Poland), Vector Laboratories (Burlingame, CA, USA), and Proteintech (Manchester, UK) and the one for mass spectrometry were from Merck (Warszawa, Poland). Animals were anesthetized with the usage of sodium pentobarbital (Biowet Puławy, Poland).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!