Endogenous Metabolite; alpha7 nicotinic receptors

Cell viability can be maintained and apoptosis effectively reduced with glycerophosphoinositide choline (0 or 70 μM, 4 days) [1].

Glycerophosphoinositide choline (subcutaneous injection, 0.2 and 100 mg/kg/h, 24 or 48 hours) significantly inhibits the release of tumor necrosis factor (TNF) from macrophages and attenuates postoperative nociceptive reflexes in female C57/Bl6 mice. 2) [2].

Cell viability assay [1]
Cell Types: rat pheochromocytoma cells PC12
Tested Concentrations: 0 or 70 μM
Incubation Duration: 4 days
Experimental Results: Cell viability was 94% at 70 μM and 83% at 0 μM. Compared with the no-treatment group, the number of cells showing DNA fragmentation (a characteristic of apoptosis) was diminished by 8.5% at 70 μM.

Animal/Disease Models: Female C57/Bl6 mouse postoperative pain model [2]
Doses: 0.2 and 100 mg/kg/h
Route of Administration: subcutaneous injection, 24 or 48 hrs (hrs (hours))
Experimental Results: Thermal allergy was diminished after surgery and reached 48 hrs (hrs (hours)) after treatment For maximum efficacy, the ED50 value of the choline dose is 1.7 mg/kg/h. Allergic responses to punctate mechanical stimulation were diminished in a dose-dependent manner with an ED50 value of 4.7 mg/kg/h at 48 hrs (hrs (hours)) but not at 24 hrs (hrs (hours)) after infusion.

[1]. Apoptosis is induced by choline deficiency in fetal brain and in PC12 cells. Brain Res Dev Brain Res. 1997 Jul 18;101(1-2):9-16.

[2]. Antinociceptive and anti-inflammatory effects of choline in a mouse model of postoperative pain. Br J Anaesth. 2010 Aug;105(2):201-7.

Treatment of rats with choline during critical periods of brain development resulted in a lasting enhancement of spatial memory in their offspring. Apoptosis is a normal phenomenon during brain development, and in certain tissues, choline availability modulates apoptosis. This study aimed to investigate whether choline availability affects apoptosis in the fetal brain and the PC12 cell line derived from rat pheochromocytoma. We fed time-mated Sprague Dawley rats a choline-deficient (CD), choline-controlled, or choline-supplemented (CS) diet for 6 days, and then prepared fetal brain sections on day 18 of embryonic development. We assessed apoptosis using terminal deoxyribonucleotide end labeling (TUNEL) to detect DNA strand breaks and count apoptotic bodies. In the hippocampus of fetuses fed CD, control, and CS diets, the proportions of TUNEL-positive cells were 15.9% (P < 0.01), 8.7%, and 7.2%, respectively. A similar negative correlation between choline intake and the number of TUNEL-positive cells was also detected in the cortical regions of these fetal brain sections. The number of apoptotic bodies in fetal brain slices was negatively correlated with maternal choline intake (the proportion of apoptotic bodies in hippocampal cells of fetuses fed CD, control, and CS diets were 6.2% (P < 0.01), 2.5%, and 1.9%, respectively). PC12 cells were cultured in DMEM/F12 medium supplemented with 70 μM choline or without choline. Compared with 70 μM choline medium (0.55%), the number of apoptotic bodies in PC12 cells cultured in 0 μM choline medium was significantly increased (1.5%; P < 0.05). In PC12 cells, TUNEL markers (DNA strand breaks) were significantly increased in choline-deficient medium (13.5%, P < 0.05) compared with choline-supplemented medium (5.0%). Furthermore, genomic DNA breaks into 200 bp internuclear body fragments were detected in choline-deficient cells. These results indicate that choline deficiency can induce apoptotic death in neuron-like cells and whole-brain cells. We believe that changes in choline availability in the brain modulate the rate of apoptosis during development. [1]

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Background: Choline is a dietary supplement that activates α7 nicotine receptors. Activation of α7 nicotine receptors reduces cytokine production in macrophages and has an analgesic effect in an inflammatory pain model. We hypothesized that systemic administration of choline would reduce the inflammatory response in macrophages and exert an analgesic effect in a mouse model of postoperative pain. Methods: We investigated the responses of wild-type and α7 nicotine receptor knockout mice to thermal and punctate pressure stimulation after model surgery. We investigated the effects of genotype and choline treatment on the binding of α-cobra venom to macrophages and their production of tumor necrosis factor (TNF). Results: Choline had a moderate analgesic effect. The ED50 of choline for inhibiting heat-induced hyperalgesia was 1.7 mg kg⁻¹ h⁻¹. The ED50 of choline for reducing the punctate pressure threshold was 4.7 mg kg⁻¹ h⁻¹. α7 nicotine receptor knockout mice did not show any change in hypersensitivity to heat or pressure, but compared with littermate control mice, there was a significant difference after treatment with 5 mg kg⁻¹ h⁻¹ choline (P<0.05, 0.01). 100 mM choline reduced the binding of α-cladosnip toxin to macrophages by 72% and reduced TNF release by up to 51% (standard deviation 11%). The inhibitory effect of choline on TNF release was not different among different genotype mice. Conclusion: Systemic choline exerts a moderate analgesic effect by activating α7 nicotine acetylcholine receptors. Its analgesic effect may not be achieved by reducing the release of macrophage TNF pathway cytokines. Although millimolecular concentrations of choline significantly inhibited TNF release, this effect was not dependent on the α7 subunit and its concentration may be higher than the systemic concentration reached in vivo. [2]

C14H32NO12P

437.37

437.166

C, 38.45; H, 7.37; N, 3.20; O, 43.90; P, 7.08

425642-32-6

Choline chloride;67-48-1; Choline bitartrate;87-67-2;Choline Fenofibrate;856676-23-8;Choline-d4 chloride;285979-70-6;Choline-d9 chloride;61037-86-3;Choline Chloride-13C3;Choline theophyllinate;4499-40-5;Glycerophosphoinositol choline;425642-32-6;Choline-d6 chloride;Choline-d13 chloride;352438-97-2;Choline-13C2 chloride;202190-49-6; 425642-32-6 (choline); 129830-95-1 (free); 425642-29-1 (potassium); 425642-30-4 (sodium)

91936944

Typically exists as solid at room temperature

8

12

8

28

401

5

P(=O)([O-])(OC[C@@H](CO)O)OC1[C@@H]([C@H](C([C@H]([C@H]1O)O)O)O)O.OCC[N+](C)(C)C

PTZZCYHESHNXFL-SECXAADESA-M

InChI=1S/C9H19O11P.C5H14NO/c10-1-3(11)2-19-21(17,18)20-9-7(15)5(13)4(12)6(14)8(9)16;1-6(2,3)4-5-7/h3-16H,1-2H2,(H,17,18);7H,4-5H2,1-3H3/q;+1/p-1/t3-,4?,5-,6+,7-,8-,9?;/m1./s1

[(2R)-2,3-dihydroxypropyl] [(2R,3R,5S,6R)-2,3,4,5,6-pentahydroxycyclohexyl] phosphate;2-hydroxyethyl(trimethyl)azanium

Plain; Glycerophosphoinositol choline; Glycerophosphoinositol choline; Plain; 425642-32-6; 3W4V4N5240; Glycerophosphoinositol (choline); UNII-3W4V4N5240; Glycerophosphoinositol choline [INCI]; D-Myo-inositol, 1-((2R)-2,3-dihydroxypropyl hydrogen phosphate), ION(1-), 2-hydroxy-N,N,N-trimethylethanaminium (1:1);

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)