Orexin a

Manufactured by Bio-Techne

Sourced in United Kingdom, United States

Orexin-A is a peptide produced by Bio-Techne. It is a key regulator of sleep-wake cycles and arousal. Orexin-A plays a crucial role in maintaining wakefulness and promoting the transition from sleep to wakefulness.

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Lab products found in correlation

Sb 334867, by Bio-Techne (8 mentions) Sb 334867 a, by Bio-Techne (3 mentions) Tcs ox2 29, by Bio-Techne (3 mentions) Capsaicin, by Merck Group (2 mentions) Prism 5, by GraphPad (1 mentions) Gf109203x, by Apexbio (1 mentions) Ly294002, by Apexbio (1 mentions) Pump 22, by Harvard Apparatus (1 mentions) Mouse anti c fos antibody, by Santa Cruz Biotechnology (1 mentions) Prism 6, by GraphPad (1 mentions) Gdp β s, by Merck Group (1 mentions) Gabazine, by Merck Group (1 mentions) Sb 674042, by Bio-Techne (1 mentions) Anti dynorphin a antibody, by Abcam (1 mentions) Dynorphin a, by Bio-Techne (1 mentions) Nrf2 antibody, by Santa Cruz Biotechnology (1 mentions) Win 55 212 2, by Merck Group (1 mentions) Naltrexone, by Merck Group (1 mentions) Am251, by Merck Group (1 mentions) Naloxone, by Merck Group (1 mentions)

23 protocols using orexin a

Orexin A and SB334867 Application

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Orexin A and N‐(2‐methyl‐6‐benzoxazolyl)‐N’‐1,5‐naphthyridin‐4‐yl urea (SB334867) were purchased from Tocris Bioscience (Bristol, United Kingdom). Orexin A was dissolved in deionized water to form stock solutions and diluted with aCSF solution to designated final concentrations. SB334867 was dissolved in dimethyl sulfoxide. The final DMSO concentration in bath solution was 0.05%, which had on effects on the firing activity of PVN neurons.

Zhou J., Ma H., Shao J., Wei Y., Zhang X., Zhang Y, & Li D. (2019). Downregulation of Orexin Receptor in Hypothalamic Paraventricular Nucleus Decreases Blood Pressure in Obese Zucker Rats. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 8(13), e011434.

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Capsaicin and Orexin-A Solubilization

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Capsaicin was dissolved in Tween 80 and ethanol solution (1 ethanol, 1 Tween 80, 8 distilled water). Orexin-A and SB-334867-A (Tocris Bioscience, Bristol, UK) were dissolved in distilled water and dimethyl sulfoxide, respectively.

Kooshki R., Abbasnejad M., Esmaeili Mahani S., Raoof M., Moeini Aghtaei M.M, & Dabiri S. (2018). Orexin-A inhibits capsaicin-induced changes in cyclooxygenase-2 and brain-derived neurotrophic factor expression in trigeminal nucleus caudalis of rats. The Korean Journal of Pain, 31(3), 174-182.

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Investigating IGF-I and Orexin A in the HDB Nucleus

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IGF-I was locally delivered in the HDB nucleus (10 nM; 0.2 μl; coordinates as above) employing a 1 μl Hamilton syringe. Besides, IGF-I was injected systemically (1 μg/g; i.p.) in other experiments. The muscarinic receptor antagonist atropine (1 mg/Kg in 0.9% NaCl i.p.) was administered 15 min before IGF-I systemic injection to assess whether IGF-I effects were due to activation of muscarinic receptors. Orexin A (Tocris, Spain) was also injected into the HDB nucleus (10 nM; 0.2 μl; coordinates as above).
In a set of experiments, we administrated human IGF-I (hIGF-I) through Alzet osmotic mini-pumps (Model 1004; USA) for chronic administration (Pre-Protech, USA; 50 g/kg/day) in ChAT-ChR2-YFP animals or the vehicle (saline solution). Pumps were implanted subcutaneously between the scapulae, following the manufacturer’s instructions. Treatment lasted 28 days. After that, animals were submitted to electrophysiological recordings as described above.

Zegarra-Valdivia J.A., Chaves-Coira I., Fernandez de Sevilla M.E., Martinez-Rachadell L., Esparza J., Torres-Aleman I, & Nuñez A. (2021). Reduced Insulin-Like Growth Factor-I Effects in the Basal Forebrain of Aging Mouse. Frontiers in Aging Neuroscience, 13, 682388.

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Orexin-A Receptor Antagonist Signaling

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Unless otherwise stated, all chemicals were purchased from Sigma Chemical, Co. (St. Louis. MO, United States). Orexin-A and SB334867 were purchased from Tocris (Bristol, United Kingdom). LY294002 and GF109203X were purchased from ApexBio (Boston, MA, United States). Results shown in the figures were representative of at least three independent experiments. All values in the figures and the text were expressed as mean ± SEM. Data analysis was performed using IBM SPSS Statistics 22.0 software and Prism 5.0. The results were analyzed by one-way or two-way analysis of variance followed by a Bonferroni post hoc test for multiple comparisons. P < 0.05 was considered statistically significant.

Liu M.F., Xue Y., Liu C., Liu Y.H., Diao H.L., Wang Y., Pan Y.P, & Chen L. (2018). Orexin-A Exerts Neuroprotective Effects via OX1R in Parkinson’s Disease. Frontiers in Neuroscience, 12, 835.

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Orexin Neurotransmission in Cocaine Addiction

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Cocaine hydrochloride (15 mg/kg in saline, Macfarlan Smith Ltd., Edinburgh, United Kingdom), orexin-A [333 pmol/μL in artificial cerebrospinal fluid (aCSF); Tocris Bioscience, United Kingdom], orexin-B [333 pmol/μL in artificial cerebrospinal fluid (aCSF); 10 ug/loci; Tocris Bioscience, United Kingdom] (Zhou et al., 2023 (link)), and TCS-OX2-29 (an OX2R antagonist; 33.3 μg/μL in 5% DMSO aCSF; Tocris Bioscience, United Kingdom) were used. The brain injection was carried out with a micro-pump (pump 22, Harvard Apparatus, Holliston, MA, United States) at a speed of 0.25 μL/min for 5 min on each side.

Jang H.B., Ahn D., Kim H.K., Guan X., Fan Y., Lee B.H, & Kim H.Y. (2023). Mediation of lateral hypothalamus orexin input to lateral habenula in the inhibitory effects of mechanical stimulation on psychomotor responses induced by cocaine. Frontiers in Molecular Neuroscience, 16, 1195939.

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Orexin-A and Capsaicin Injection Protocol

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Orexin-A (Tocris Bioscience, Bristol, UK) was added to synthetic CSF. The orexin-1 receptor (OX1R) antagonist SB-334867-A (Tocris Bioscience) was dissolved in dimethyl sulfoxide and diluted in synthetic CSF. The drugs were injected into the RVM with a polyethylene syringe (volume: 1 μL). Capsaicin (Sigma-Aldrich, St. Louis, MO) was added to a solution containing 80% distilled water, 10% ethanol, and 10% Tween 80. Capsaicin (100 μg/10 μL) was intradentally administrated in the rats’ mandibular incisors.

Shahsavari F., Abbasnejad M., Esmaeili-Mahani S, & Raoof M. (2022). The ability of orexin-A to modify pain-induced cyclooxygenase-2 and brain-derived neurotrophic factor expression is associated with its ability to inhibit capsaicin-induced pulpal nociception in rats. The Korean Journal of Pain, 35(3), 261-270.

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Orexin-A and SB334867 in PTZ Seizure

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Orexin-A (cat, 1455; OX1R and OX2R agonist) and SB334867 (cat, 1960; OX1R antagonist) from Tocris Bioscience and PTZ (cat, P6500) from Sigma Aldrich was used in this research. Lidocaine 2% was purchased from local drug market in IRAN.

Akbari N., Salmani M.E., Goudarzvand M., LashkarBoluki T., Goudarzi I, & Abrari K. (2014). Unilateral Hypothalamus Inactivation Prevents PTZ Kindling Development through Hippocampal Orexin Receptor 1 Modulation. Basic and Clinical Neuroscience, 5(1), 66-73.

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Orexin-A and SB-334867 Neuromodulation Protocol

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Orexin-A and SB-334867 were purchased from Tocris (Bristol, United Kingdom). SCH-22390 was purchased from Sigma-Aldrich (St Louis, MO, United States) Mouse anti-c-fos antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States) and Cy3-binding goat anti-mouse antibody purchased from Jackson ImmunoResearch (West Grove, PA, United States). The data were analyzed using Prism 6 (GraphPad Software, San Diego, CA, United States), and the results were expressed as the mean ± SD. Paired Student’s t-test was used to analyze changes in discharge rate before and after treatment. Student’s t-test was used to compare differences between two groups, and one-way or two-way analysis of variance followed by Bonferroni post hoc test were used to compare differences among three or more groups. P < 0.05 was considered statistically significant.

Jin T., Jiang Z., Luan X., Qu Z., Guo F., Gao S., Xu L, & Sun X. (2020). Exogenous Orexin-A Microinjected Into Central Nucleus of the Amygdala Modulates Feeding and Gastric Motility in Rats. Frontiers in Neuroscience, 14, 274.

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Electrophysiological Assay of Orexin Receptor Antagonists

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Orexin A, N-(2-methyl-6-benzoxazolyl)-N-1,5-naphthyridin-4-yl urea (SB-334867), and (2S)-1-(3,4-dihydro-6,7-dimethoxy-2(1H)-isoquinolinyl)-3,3-dimethyl-2-[(4-pyridinylmethyl)amino]-1-butanone hydrochloride (TCS-OX2-29) were purchased from Tocris. CNQX, AP5, gabazine, and GDP-β-s were obtained from Sigma Aldrich. For the electrophysiological experiments, all reagents, except SB-334867, were dissolved in deionized water to form stock solutions and diluted with aCSF solution. SB-334867 was dissolved in dimethyl sulfoxide (DMSO), and the final concentration of DMSO in the bath solution was 0.05% which showed no effects on the firing activity of the vehicle control neurons.

Zhou J.J., Yuan F., Zhang Y, & Li D.P. (2015). Upregulation of orexin receptor in paraventricular nucleus promotes sympathetic outflow in obese Zucker rats. Neuropharmacology, 99, 481-490.

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Intrathecal administration of orexin, dynorphin, and SB674042 in von Frey tests

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Orexin A, SB674042, and dynorphin A were purchased from Tocris Bioscience (Bristol, UK), and the anti-dynorphin A antibody was purchased from Abcam (Cambridge, UK). SB674042 was dissolved in dimethyl sulfoxide (DMSO) and the final solution contained 6% DMSO. The remaining samples were dissolved in saline solution. The amount of intrathecally administered drug was 5 µg in a volume of 5 µL and all PE-10 tubes were flushed with 10 µL of saline [21 (link)–24 (link, link, link)]. All drugs were injected 30 minutes before the von Frey tests.

Youn D.H., Jun J., Kim T.W, & Park K. (2022). Spinal orexin A attenuates opioid-induced mechanical hypersensitivity in the rat. The Korean Journal of Pain, 35(4), 433-439.

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