Gaboxadol

Manufactured by Merck Group

Sourced in Australia

Gaboxadol is a research tool used in experimental studies. It functions as a selective agonist for the GABA-A receptor. This compound is utilized by researchers to investigate the role of GABA-A receptor activation in various biological processes.

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Gaboxadol hydrochloride

16 protocols using gaboxadol

Gaboxadol-Enriched Paraquat Exposure

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Gaboxadol hydrochloride (Sigma-Aldrich, St. Louis, MO) was dissolved in water and added to melted cornmeal food to a final concentration of 0.1–0.2 mg/mL. Flies were flipped onto Gaboxadol-containing food for 3 days prior to paraquat injection and remained on Gaboxadol-containing food postinjection. Control food was made by adding the appropriate amount of vehicle alone (H₂O) to melted cornmeal food.

Hill V.M., O’Connor R.M., Sissoko G.B., Irobunda I.S., Leong S., Canman J.C., Stavropoulos N, & Shirasu-Hiza M. (2018). A bidirectional relationship between sleep and oxidative stress in Drosophila. PLoS Biology, 16(7), e2005206.

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Zebrafish Neuropharmacology Protocol

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All reagents were prepared fresh on the day of the experiment and dissolved in E3 to create working solutions to be added to the experimental chamber, and compared to the addition of carrier-only controls. Gaboxadol (Sigma T101), MS222 (Sigma), mepyramine (Sigma P5514), promethazine (Sigma P4651), carbachol (Sigma C4382), eserine (Sigma E8375) and methoctramine (Sigma M105) were all dissolved in E3. H6408 (Bachem; H-6408.0001) was prepared with double-distilled (dd) H₂0 (ddH₂0) at 1 mM with a working concentration at 10 μM. Zolpidem (Sanofis Pharmaceuticals; active ingredient of the prevalent sleep drug Ambien) was a gift from S. Nishino, and stock solutions were prepared in DMSO. Effective concentrations and fish age (ranging from 7 dpf–14dpf) were chosen based on published data^{47 (link),48 (link)} or dose–response experiments driving behavioural sleep in the Viewpoint behavioural tracking system (Supplementary Table 1). At least two independent tests for all drug dose–responses were performed.
Injected MCH peptide (Phoenix Pharmaceuticals; 070–47, lot no. 429808) was prepared as a 1 mM working solution in ddH₂0 supplemented with phenol red solution (Sigma; P0290) to confirm injection (see Extended Data Fig. 7h). Intracerebroventricular pulsatile injections (FemtoJet Microinjector, Eppendorf) were performed with zPSG larvae mounted in agarose.

Leung L.C., Wang G.X., Madelaine R., Skariah G., Kawakami K., Deisseroth K., Urban A.E, & Mourrain P. (2019). Neural signatures of sleep in zebrafish. Nature, 571(7764), 198-204.

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Modulating Neural Activity with Pharmacological Agents

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Neural activity was modulated using pentylenetetrazole (PTZ, Sigma, P6500), muscimol (Sigma, M1523), caffeine (Sigma, C0750), 4-aminopyridine (4-AP, Sigma, 275875), and gaboxadol (Sigma, T101). Stocks were prepared in molecular grade water. To test the impact on turn bias, larvae were treated with each drug diluted in E3 to working concentrations. The CP assay was conducted as previously mentioned. Drugs were replaced daily for all experiments. At 4 dpf, the drug(s) were removed, and fresh E3 was used for the rest of the experiment.

Hageter J., Starkey J, & Horstick E.J. (2023). Thalamic regulation of a visual critical period and motor behavior. Cell reports, 42(4), 112287.

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Recombinant Expression of Human GABA Receptor

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Human ρ1 cDNA subcloned into pcDNA1.1 (Invitrogen, San Diego, CA, USA) was kindly provided by Dr. George Uh1 (National Institute for Drug Abuse, Baltimore, MD, USA).
(GABA, 5-aminovaleric acid, β-alanine, glycine, isoguvacine, 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) also known as Gaboxadol, were purchased from Sigma-Aldrich (Sigm-Aldrich Pty Ltd, Castle Hill, NSW 1765 Australia). 1,2,5,6-Tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA) [31 (link)], trans-aminocrotonic acid (TACA) and cis-aminocrotonic acid (CACA) [32 (link)], and 2-aminoethylmethane thiosulfonate (MTSEA) [33 (link)] were prepared as previously reported.

Naffaa M.M., Absalom N., Solomon V.R., Chebib M., Hibbs D.E, & Hanrahan J.R. (2016). Investigating the Role of Loop C Hydrophilic Residue ‘T244’ in the Binding Site of ρ1 GABAC Receptors via Site Mutation and Partial Agonism. PLoS ONE, 11(5), e0156618.

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GABAA Receptor Modulation: Mechanisms

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Drugs were injected i.h. at a volume of 0.25 μl per side at a rate of 0.15 μl min⁻¹ or i.p. at a volume of 0.2 ml. Extrasynaptic GABA_A receptors were activated by gaboxadol (varying doses, indicated in corresponding figures, dissolved in 0.9% saline; vol/vol, Sigma-Aldrich), PKCβII was inhibited by PKCβII peptide inhibitor I trifluoroacetate salt (0.25 μg per hippocampus, dissolved in artificial cerebrospinal fluid; Sigma-Aldrich) and PKCδ was inhibited by myristoyl (N-terminal) SFNSYELGSL peptide (0.5 μg per hippocampus, dissolved in artificial cerebrospinal fluid; GenScript).

Jovasevic V., Corcoran K.A., Leaderbrand K., Yamawaki N., Guedea A.L., Chen H.J., Shepherd G.M, & Radulovic J. (2015). GABAergic mechanisms regulated by miR-33 encode state-dependent fear. Nature neuroscience, 18(9), 1265-1271.

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Sleep Induction in Drosophila

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For sleep induction experiments, flies were placed overnight on 1% agar and 5% sucrose food laced with Gaboxadol (0.2 mg/ml; Sigma-Aldrich). A little blue food coloring was added to the food, so flies who had recently fed could be readily identified by their blue bellies.

van Alphen B., Semenza E.R., Yap M., van Swinderen B, & Allada R. (2021). A deep sleep stage in Drosophila with a functional role in waste clearance. Science Advances, 7(4), eabc2999.

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Intrahippocampal Gaboxadol Administration

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Gaboxadol (0.25 or 0.5 μg, dissolved in artificial cerebrospinal fluid (ACSF), Sigma-Aldrich) was injected intrahippocampally (i.h.), at a volume of 0.5 μl per side, at a rate of 0.15 μl/min.

Jovasevic V, & Radulovic J. (2021). High ethanol preference and dissociated memory are co-occurring phenotypes associated with hippocampal GABAAR-δ receptor levels. Neurobiology of learning and memory, 183, 107459.

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Gaboxadol Pharmacokinetics and Exposure

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Animals were injected with Gaboxadol (3 mg/kg and 10 mg/kg i.p.; Cat.#: T101, Sigma-Aldrich, Darmstadt, GER) or vehicle solution (99.7% of a 0.9% saline solution + 0.3% Tween20; Cat.#: 11332465001, Sigma-Aldrich). Dosing was randomized using a latin-based square design, with each animal receiving every compound/vehicle for within-subject comparison. Dosing was performed fifteen minutes before starting the data acquisition, in line with the previously established pharmacokinetics (data not shown). No blinding was performed. The duration of the washout phase between dosings was 48 h. Blood samples were collected from the tail vein approximately two hours after dosing and exposures were confirmed by Hoffmann-La Roche bioanalytics.

Janz P., Bainier M., Marashli S., Schoenenberger P., Valencia M, & Redondo R.L. (2022). Neurexin1α knockout rats display oscillatory abnormalities and sensory processing deficits back-translating key endophenotypes of psychiatric disorders. Translational Psychiatry, 12, 455.

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Anesthetic Cocktail Preparation and Delivery

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Injectable ethanol (Decon Labs Inc., #2701) solutions were prepared in 0.9% saline (15% and 20%, v/v). Gaboxadol (55 mg/kg; Sigma‐Aldrich, #T101) and ketamine (175 mg/kg; Sigma‐Aldrich, #K2753) were dissolved in 0.9% saline and injected at 0.1 mL/10 g of body weight. Zolpidem (60 mg/kg; Tocris Bioscience, #0655) was freshly prepared as a suspension in saline with 3–4 drops of Tween‐80 (Sigma‐Aldrich, #P1754) and administered daily in a volume of 0.1 mL/10 g of body weight. All drugs were administered interperitoneally (i.p.).

Plasil S.L., Farris S.P., Blednov Y., Mayfield R.D., Mangieri R.A., Nwokeji U.J., Aziz H.C., Lambeth P.S., Harris R.A, & Homanics G.E. (2024). Mutation of novel ethanol‐responsive lncRNA Gm41261 impacts ethanol‐related behavioral responses in mice. Genes, Brain, and Behavior, 23(1), e12886.

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Intraperitoneal Drug Administration in Rodents

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Injectable ethanol (Aaper Alcohol and Chemical, Shelbyville, KY) solutions were prepared in 0.9% saline (20%, v/v). Gaboxadol (Sigma-Aldrich, St. Louis, MO), diazepam (Sigma-Aldrich), and pentobarbital (Sigma-Aldrich RBI, Natick, MA) were dissolved in 0.9% saline. All drugs were injected intraperitoneally (i.p.) at 0.01 ml/g of body weight.

Blednov Y.A., Black M., Benavidez J.M., Da Costa A., Mayfield J, & Harris R.A. (2017). Sedative and motor incoordination effects of ethanol in mice lacking CD14, TLR2, TLR4, or MyD88. Alcoholism, clinical and experimental research, 41(3), 531-540.

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