The drugs administered during the experiments were amiloride hydrochloride hydrate (molecular weight [MW] 266.09; Sigma-Aldrich, USA), tizanidine hydrochloride (MW 290.17; Sigma-Aldrich, USA) and yohimbine hydrochloride (MW 390.90; Sigma-Aldrich, USA). All drugs were dissolved in normal saline. Amiloride and tizanidine were each intrathecally administered 10 min before the formalin test. To assess the dose dependency and time courses of the antinociceptive action of intrathecal amiloride, tizanidine and coadministered amiloride-tizanidine, the animals were randomly assigned to different groups receiving intrathecal injections of different doses of amiloride (12.5 μg, 25 μg, 50 μg, 100 μg), tizanidine (0.5 μg, 1 μg, 2.5 μg, 5 μg) or amiloride plus tizanidine 10 min before formalin injection. In the control group, intrathecal saline was given. Yohimbine was intrathecally administered 10 min before 5 μg tizanidine, 100 μg amiloride or the dose that resulted in 50% of the saline response (ED50)/2 of amiloride + tizanidine ED50/2 mixture. The agents were delivered using a microsyringe in a total volume of 10 mL, followed by 10 mL of saline to flush the catheter.
Yohimbine hydrochloride
Manufactured by Merck Group
Sourced in United States, Germany
Yohimbine hydrochloride is a chemical compound used in various laboratory applications. It is a crystalline powder that is soluble in water and organic solvents. Yohimbine hydrochloride is commonly used in research and scientific experiments, but its specific applications and functions may vary depending on the context and intended use.
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Lab products found in correlation
Prazosin hydrochloride, by Merck Group (5 mentions)
Clonidine hydrochloride, by Merck Group (2 mentions)
Nicotine hydrogen tartrate, by Merck Group (2 mentions)
Serotonin hydrochloride, by Merck Group (2 mentions)
Tyramine hydrochloride, by Merck Group (2 mentions)
Octopamine hydrochloride, by Merck Group (2 mentions)
Naltrexone hydrochloride, by Merck Group (2 mentions)
Tizanidine hydrochloride, by Merck Group (1 mentions)
Amiloride hydrochloride hydrate, by Merck Group (1 mentions)
Domosedan, by Pfizer (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Human insulin, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium f12, by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Propranolol, by Merck Group (1 mentions)
Foetal bovine serum (fbs), by Harvard Bioscience (1 mentions)
Isoprenaline hydrochloride, by Merck Group (1 mentions)
Itaq universal sybr green supermix, by Bio-Rad (1 mentions)
30 protocols using yohimbine hydrochloride
1
Intrathecal Antinociceptive Synergism
2
Testosterone Analysis of Canine Semen
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Blood samples were collected from the jugular vein in heparinized tubes between 10:00 h and 11.00 h. The collected blood was centrifuged at 1500× g for 15 min, the plasma separated, and stored at −20 °C until required for testosterone analysis.
Ejaculates were collected by the transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) [43 (link)]. Animals were anesthetized by 0.5 mg/kg intravenous ketamine hydrochloride (Imalgene-1000) (Rhône Mérieux, Lyon, France), 50 µg/kg detomidine (Domosedan) (Pfizer Inc., Amboise Cedex, France), and 0.5 mg/kg tiletamine-zolazepan (Zoletil-100) (Virbac España SA, Barcelona, Spain) Anesthesia was maintained via isoflurane (Isobavet) (Intervet Schering-Plough Animal Health) and later reversed using 0.7 mg/kg yohimbine hydrochloride (half intravenous and half intramuscular) (Sigma, Zwijndrecht, The Netherlands). The time between semen collections for each animal was 14–15 days.
Ejaculates were collected by the transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) [43 (link)]. Animals were anesthetized by 0.5 mg/kg intravenous ketamine hydrochloride (Imalgene-1000) (Rhône Mérieux, Lyon, France), 50 µg/kg detomidine (Domosedan) (Pfizer Inc., Amboise Cedex, France), and 0.5 mg/kg tiletamine-zolazepan (Zoletil-100) (Virbac España SA, Barcelona, Spain) Anesthesia was maintained via isoflurane (Isobavet) (Intervet Schering-Plough Animal Health) and later reversed using 0.7 mg/kg yohimbine hydrochloride (half intravenous and half intramuscular) (Sigma, Zwijndrecht, The Netherlands). The time between semen collections for each animal was 14–15 days.
3
Molecular Mechanisms of Adrenergic Signaling
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(−)-Adrenaline bitartrate, (−)-isoprenaline hydrochloride, prazosin hydrochloride, yohimbine hydrochloride, (±)-propranolol, α-methylDOPA, Dulbecco’s Modified Eagle’s Medium/F-12 supplemented with NaHCO3, Dulbecco’s Modified Eagle’s Medium with NaHCO3 and stable L-glutamine, epidermal growth factor, human insulin, hydrocortisone, ICI 118,551, penicillin/streptomycin and HPLC-ECD standards from the highest purity available, were from Sigma-Aldrich (Sintra, Portugal). ITaqTM Universal SYBR Green supermix was from Biorad (Amadora, Portugal). Foetal bovine serum was from Biochrom and L-glutamine was from Gibco (Biotecnómica, São Mamede, Portugal).
Goat anti-mouse IgG conjugated with Alexa Fluor 488 (a11029) and mouse monoclonal anti-phenylethanolamine N-methyltransferase (MA5-25530) were from Thermo Fisher Scientific (Loures, Portugal). Rabbit polyclonal anti-β2-adrenoceptor (13096-1-AP) was from Proteintech (Rosemont, IL, USA). Goat anti-rabbit IgG conjugated with Alexa Fluor 594 (ab150092) and rabbit monoclonal anti-tyrosine hydroxylase (ab137869) were purchased from Abcam (Cambridge, UK). Goat anti-rabbit IgG conjugated with horseradish peroxidase (sc-2004) was from Santa Cruz Biotechnology Inc. (Frilabo, Maia, Portugal).
Goat anti-mouse IgG conjugated with Alexa Fluor 488 (a11029) and mouse monoclonal anti-phenylethanolamine N-methyltransferase (MA5-25530) were from Thermo Fisher Scientific (Loures, Portugal). Rabbit polyclonal anti-β2-adrenoceptor (13096-1-AP) was from Proteintech (Rosemont, IL, USA). Goat anti-rabbit IgG conjugated with Alexa Fluor 594 (ab150092) and rabbit monoclonal anti-tyrosine hydroxylase (ab137869) were purchased from Abcam (Cambridge, UK). Goat anti-rabbit IgG conjugated with horseradish peroxidase (sc-2004) was from Santa Cruz Biotechnology Inc. (Frilabo, Maia, Portugal).
4
Preparation of Yohimbine and WAY100635
5
Subcutaneous and Oral Cytotoxin Delivery
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In in vivo assays, except for the LD50 experiments, where the doses will be presented in the test description, the following compounds were used: 40 μg/kg (in 200 µL of PBS) of CTX [13 (link)] or 54 μg/kg (in 200 µL of PBS) of CTX:SBA-15 administered by subcutaneous (s.c.) or oral route (p.o.); the formyl peptide receptor antagonist, Boc2 (butoxycarbonil-Phe-Leu-Phe-Leu-Phe, #Phoenix Pharmaceutical Inc, USA) at 10 μg/kg (in 100 µL of sterile saline), the muscarinic receptors antagonist, atropine sulphate (Sigma #5908996) at 10 mg/kg (in 100 µL of sterile saline), and the α-adrenergic receptor antagonist, yohimbine hydrochloride (Sigma #y3125) at 2 mg/kg (in 100 µL of sterile saline), were intraperitoneally (i.p.) administered. All the antagonists were administered 30 min prior the treatment with CTX or CTX:SBA-15.
6
Alcohol, Tamoxifen, and Yohimbine Protocol
7
Intrathecal α2-Receptor Modulation
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The drugs used were clonidine hydrochloride (α2-agonist), yohimbine hydrochloride (α2-antagonist/5HT agonist) and methysergide maleate (5HT antagonist) (all from Sigma). The aim of using methysergide maleate was to determine any interaction of 5-HT system with α2-antagonist (yohimbine). clonidine hydrochloride and methysergide maleate were dissolved in 0.9% saline. Yohimbine was dissolved in dimethyl sulphoxide (DMSO). Drugs or vehicles were administered intrathecally in a volume of 100 μL using a 30-gauge needle. The dosages used were based on preliminary investigations from our laboratory. Fresh preparations of drugs were always used.
8
Alcohol Drinking and Yohimbine Effects
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The alcohol drinking solution 10% (v/v) was prepared by diluting 95% alcohol (F.L. Carsetti, Camerino, Italy) with tap water. Yohimbine hydrochloride (17-hydroxyyohimban-16-carboxylic acid methyl ester hydrochloride) was purchased from Sigma-Aldrich, Italy, dissolved in sterile distilled water, and administered intraperitoneally (i.p.) at 0.0, 0.312, 0.625, and 1.25 mg/kg in a 1 ml/kg injection volume 30 min before the drug tests (Lê et al., 2005 (link); Marinelli et al., 2007 (link); Ayanwuyi et al., 2013 (link)).
9
Rat Ghrelin and Norepinephrine Regulation
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Rat UCN1 and rat acylated ghrelin were purchased from Peptide Institute, Inc. (Osaka, Japan). Yohimbine hydrochloride, a selective α2‐AR antagonist, was purchased from Sigma‐Aldrich Chemical Co. (St. Louis, MO, USA). The GHS‐R1a antagonist, [D‐Lys3]‐GHRP‐6, was purchased from Bachem, Inc. (Torrance, CA, USA). These compounds were dissolved in saline when injected by IP and intravenous (IV) routes. Rikkunshito, a Japanese traditional Kampo medicine, was supplied from Tsumura & Co. in the form of a powdered extract obtained by spray‐drying a hot water extract mixture of the following eight crude drugs: Atractylodis lanceae rhizoma (4.0 g), Ginseng radix (4.0 g), Pinelliae tuber (4.0 g), Poria (4.0 g), Zizyphi fructus (2.0 g), Citri unshiu pericarpium (2.0 g), Glycyrrhizae radix (1.0 g), and Zingiberis rhizoma (0.5 g). Rikkunshito was dissolved in distilled water when injected by orogastric administration (PO). Other analytical reagents included commercially available highest‐purity products.
Treatments were performed on unanesthetized and lightly hand‐restrained rats using the following volumes: 10 μL/rat for ICV injection, 1 mL/kg for IP or IV injection through the tail vein, and 10 mL/kg for PO.
Treatments were performed on unanesthetized and lightly hand‐restrained rats using the following volumes: 10 μL/rat for ICV injection, 1 mL/kg for IP or IV injection through the tail vein, and 10 mL/kg for PO.
10
Cigarette Smoke Extract Preparation
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Nicotine hydrogen tartrate (Sigma, St Louis, MO) was dissolved in sterile saline and adjusted to pH 7.2–7.4. All nicotine doses were calculated as free base. CSE was created by bubbling smoke from commercial cigarettes (Camel unfiltered, RJ Reynolds) through sterile saline (26 (link), 29 (link)). Briefly, eight cigarettes were smoked through 35 ml of saline solution (35-ml puffs over 2 s, repeated every 30 s) and the final solution was adjusted to pH 7.2–7.4. The CSE solution was prepared each day immediately before experimental testing in order to minimize differences resulting from differential stability of the constituents including nicotine (29 (link)). All CSE doses were defined by the solution’s nicotine content, which was analyzed by GC–MS (Finnigan Trace MS with Trace GC 2000 series, UCI Mass Spectrometry Facility). To ensure consistent nicotine concentration within CSE, periodic batches were sent to an outside facility (UCSF Clinical Pharmacology Laboratory) to confirm nicotine content. Yohimbine hydrochloride (Sigma-Aldrich) was dissolved in double distilled water.
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