WT mice used in this study were on the C57BL/6J background and from The Jackson Laboratory (Bar Harbor, ME, USA). Choline transporter heterozygous deficient (ChTHET) mouse breeders were provided by Dr. Randy Blakely (Florida Atlantic University, Boca Raton, FL, USA). Twelve-month-old female mice/15-month old male mice were treated daily for 6 weeks via intraperitoneal injection (i.p.) with galantamine hydrobromide (Sigma-Aldrich, St. Louis, MO, USA; G1660) at 2.5 mg/kg/day or vehicle, and were euthanized at 13.5 and 16.5 months of age. Fourteen-week-old female/male mice were treated daily for 6 weeks with galantamine hydrobromide or pyridostigmine bromide (Sigma-Aldrich; P9797) at 2.5 mg/kg/day and 1 mg/kg/day, respectively. Pyridostigmine was administered via mini-osmotic pumps implanted subcutaneously (Durect Corporation, Cupertino, CA, USA; 0000298) to reduce biohazard concerns.(38 (link)) All procedures were approved by the Institutional Animal Care and Use Committee at Baylor College of Medicine.
Galantamine hydrobromide
Manufactured by Merck Group
Sourced in United States, Germany, Italy
Galantamine hydrobromide is a laboratory-grade chemical compound used in various research and analytical applications. It is the hydrobromide salt of galantamine, a naturally occurring alkaloid. The compound is primarily used as a reference standard or analytical reagent in research, testing, and quality control processes.
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Lab products found in correlation
Acetylthiocholine iodide, by Merck Group (7 mentions)
Acetylcholinesterase, by Merck Group (5 mentions)
Donepezil hydrochloride, by Merck Group (4 mentions)
Gallic acid, by Merck Group (4 mentions)
5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (3 mentions)
Butyrylthiocholine iodide, by Merck Group (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
5 5 dithiobis 2 nitrobenzoic acid dtnb, by Merck Group (2 mentions)
2 4 6 tripyridyl s triazine tptz, by Merck Group (2 mentions)
Butyrylcholinesterase, by Merck Group (2 mentions)
Ascorbic acid, by Merck Group (2 mentions)
Trichloroacetic acid, by Merck Group (2 mentions)
Quercetin, by Merck Group (2 mentions)
Bovine brain extract, by Merck Group (1 mentions)
Sulfanilamide, by Merck Group (1 mentions)
Rosmarinic acid, by Merck Group (1 mentions)
Acetylcholinesterase ache from electric eel, by Merck Group (1 mentions)
Potassium ferricyanide, by Merck Group (1 mentions)
Folin ciocalteu s phenol reagent, by Merck Group (1 mentions)
Tacrine hydrochloride, by Merck Group (1 mentions)
19 protocols using galantamine hydrobromide
1
Galantamine and Pyridostigmine Treatment in Mice
2
Acetylcholinesterase Inhibition and Antioxidant Assays
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Acetylcholinesterase (AChE) from electric eel (Type VI-S, EC 3.1.17.), acetylthiocholine iodide (ATChI), bovine brain extract (Folch type VII), 3-(2-Pyridyl)-5,6-diphenyl-1,2,4-triazine-4′,4′′-disulfonic acid (ferrozine), 2,2-diphenyl-1-picrylhydrazyl (DPPH•), 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), galantamine hydrobromide, sulfanilamide, and rosmarinic acid (96%) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Aluminium chloride, ethylenediaminetetraacetic acid (EDTA), and hexamethylenetetramine were purchased from Kemika (Zagreb, Croatia). Butylhydroxytoluene (BHT, ≥99%), luteolin (≥97%), N-(1-naphthyl)ethylenediamine, sodium nitroprusside, and trolox (≥98%) were purchased from Fluka (Buchs, Switzerland). Folin-Ciocalteu's phenol reagent and potassium ferricyanide were obtained from Merck (Darmstadt, Germany). All other chemicals and solvents used were of analytical grade.
3
Acetylcholinesterase Inhibition Assay
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Recombinant human acetylcholinesterase (hAChE), Torpedo californica acetylcholinesterase (tcAChE), acetylthiocholine iodide (ATCI), 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB), all-trans-retinoic acid, thiazolyl blue tetrazolium bromide (MTT), isopropanol, dimethyl sulfoxide (DMSO), and the drugs donepezil hydrochloride, MW = 415.95 g/mol; tacrine hydrochloride, MW = 234.72; berberine hydrochloride, MW = 371.82 g/mol; and galantamine hydrobromide, MW = 368.27 g/mol, were all purchased from Sigma-Aldrich (Poole, UK). Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), albumin, and poly-D-lysine (PDL) were purchased from Thermofisher Scientific (Rochester, UK). Unless specified differently, all other chemicals were from Sigma-Aldrich (Poole, UK).
4
Zebrafish Behavioral Modulation Protocol
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Before rapamycin injections, the zebrafish were fasted for 24 h. The injection solution consisted of 5% stock solution and 95% sterile saline solution. The control injection solution consisted of 5% DMSO and 95% sterile saline. Rapamycin was intraperitoneally injected (50 μg/g). Prior to injection, the zebrafish were anesthetized with tricaine. Before being subjected to behavioural or cognitive experiments, zebrafish were immersed in water tanks containing galantamine hydrobromide (50 μg/L, Sigma-Aldrich), fluoxetine hydrochloride (50 μg/L, Sigma-Aldrich), or GR55562 dihydrochloride (100 μg/L, Abcam, Waltham, MA, USA) for 30, 60, and 90 min respectively.
5
Cholinesterase Enzyme Assay Protocol
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Acetylcholinesterase (AChE) from electric eel (C3389-2KU), butyrylcholinesterase (BuChE) from horse serum (E.C.3.1.1.8, Sigma St. Louis, MO, USA), acetylthiocholine iodide and butyrylthiocholine chloride (Sigma St. Louis, MO, USA), 5,5′-dithiobis [2-nitrobenzoic acid] (DTNB) (Sigma), galantamine hydrobromide from Lycoris sp. (Sigma St. Louis, MO, USA). Instrumentation: (Shimadzu, UV-1700).
6
Antioxidant Capacity Evaluation Protocol
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Alpha-tocopherol, β-nicotinamide-adenine dinucleotide (NADH), 2,2′-diphenyl-1-picrylhydrazyl (DPPH), 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB), acetylthiocholine iodide (ATChI), AChE, galantamine hydrobromide, gallic acid, and phenazine methosulfate (PMS) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). 2,2′-azino-bis(3-ethylbenzothioazoline-6-sulphonic acid) diammonium salt (ABTS), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), catechin, nitroblue tetrazolium (NBT), and quercetin were purchased from Fluka Chemical Co. (Buchs, Switzerland). 2,4,6-tripyridyl-s-triazine (TPTZ) was purchased from Merck Chemical Co. (Darmstadt, Germany). All other chemicals were analytical grade.
7
Cholinesterase Inhibition Assay Protocol
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Plant materials were sourced from Divya Pharmacy, Haridwar, India, and were independently certified by Council of Scientific and Industrial Research—National Institute of Science Communication and Information Resources (CSIR—NISCAIR), New Delhi, India. Identification voucher numbers of B. Monnieri, W. somnifera, C. Pluricaulis, C. Paniculatus and T. cordifolia were NISCAIR/RHMD/Consult/2019/3453-54-32, 3453-54-15, 3453-54-167, 3453-54-120, and 3453-54-63, respectively. Acetylthiocholine chloride (ATCC), acetylcholinesterase from Electrophorus electricus (electric eel) (type VI-S, lyophilized powder, CAS Number: 9000-81-1), 5, 5-dithiobis [2-nitrobenzoic acid] (DTNB), and galantamine hydrobromide were purchased from Sigma (India). All other reagents were of analytical grade and purchased locally.
8
Synthesis and Characterization of NMZ
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All chemicals and reagents were purchased from commercial sources in the highest purity and were used as received. RIV-1061 (NMZ) was synthesized in-house according to our previous literature [39 (link)]. 4-Hydroxynonenal and 2-oxo-nonenal were purchased from Cayman Chemical (Michigan, USA). Donepezil hydrochloride, arecoline hydrobromide, memantine hydrochloride, galantamine hydrobromide, chlormethiazole hydrochloride and lipopolysaccharides (from Escherichia coli O111:B4) were all purchased from Sigma (Missouri, USA). All data was generated, reported and analyzed as the mean ± S.E.M. using ANOVA with Dunnett or Tukey's multi-comparison analysis using Graph-Pad Prism version 7.0.
9
Preparation of Alzheimer's Compounds
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Donepezil Hydrochloride (obtained from Sigma Aldrich, St. Louis, MO, USA), Galantamine Hydrobromide (obtained from Sigma Aldrich, St. Louis, MO, USA), Queuine Hydrochloride (obtained from Santa-Cruz) and Etoperidone Hydrochloride (obtained from Santa-Cruz) were all dissolved in dimethyl sulfoxide (DMSO). Thiamine Hydrochloride (obtained from Sigma Aldrich, St. Louis, MO, USA) was dissolved in double distilled water (ddH2O) Galantamine we purchased was extracted from Lycoris sp. plant.
10
Preparation of Common Pharmacological Agents
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Acetylcholine chloride, PHA-543613 (PHA), lipopolysaccharides (LPS), methyllycaconitine citrate salt (MLA), donepezil hydrochloride (DPZ) and galantamine hydrobromide (GAL) were purchased from Sigma (St. Louis, MO, USA). Proper amount of chemical was dissolved in Mili Q water (Merck, Darmstadt, Germany) and stored at −20 °C.
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