Pretreatment with 50 mM 4-phenylpyridine reduced the IC50 value of MPTP (the concentration that inhibits 50% of the convulsion amplitude) in the phrenic nerve-diaphragm of mice from 53 mM to 18 mM, and the IC50 value of d-tubocurarine from 0.7 mM to 0.3 mM[2].

MPTP can be used to construct a Parkinson's disease model in animal models. MPTP can simulate naturally occurring neurodegeneration and can be used to study dopaminergic neuronal degeneration, mitochondrial dysfunction, and neuroinflammation. After injection, MPTP is rapidly metabolized to MPP+, and its serum half-life in sheep is about 6 days. Induction of Parkinson's disease model [4][5][6][7] Background: MPTP can freely cross the blood-brain barrier into the brain and is metabolized to MPP+, its active and toxic form, by monoamine oxidase B (MAO-B) in astrocytes. MPP+ is taken up by dopaminergic neurons via dopamine transporter (DAT), blocking complex I in the mitochondrial electron transport chain, triggering oxidative stress and mitochondrial dysfunction, and ultimately leading to neuronal apoptosis. Parkinson's disease is caused by the loss of neurons in the substantia nigra (the part of the substantia nigra striatum that produces dopamine). The toxic effects of MPTP can lead to the death of dopaminergic neurons in the substantia nigra, thereby causing symptoms similar to Parkinson's disease. Specific modeling method: Mice: C57BL/6 • Male • 8-12 weeks old (cycle: 2 weeks), older mice may be more sensitive. Administration route: Acute model: 14-20 mg/kg • Intraperitoneal injection • 4 times daily, 2 hours apart. Subacute model: 20-30 mg/kg • Intraperitoneal injection • Once daily for 5 consecutive days. Note 1: After administration, observe whether mice exhibit symptoms such as decreased activity, unsteady gait, convulsions, ruffled fur, and frequent urination. These behaviors may last for 24-48 hours, after which the mice generally return to normal. 2. MPTP is usually sold in the form of MPTP hydrochloride. The molecular weight of MPTP hydrochloride is 209.7. Therefore, when preparing the injection solution, it is recommended to consider the presence of hydrochloride (HCl). The molecular weight of HCl is 35.4, accounting for 17% of the total MPTP. Therefore, if a 20 mg/kg dose of MPTP is prepared, the actual dose of MPTP hydrochloride administered is 20 mg/kg * 1.17% = 23.4 mg/kg. 3. If multiple injections are given within a day, it is best to alternate between the two sides. If daily injections are given, they should be administered at the same time. Before each injection, the mouse weight should be weighed and the dose adjusted. 4. Mice with Parkinson's disease models may not exhibit the behavioral deficits of Parkinson's disease. Individual differences may exist among mice, so the success rate of model establishment is usually difficult to achieve 100%. Therefore, in MPTP mouse studies, the primary focus should be on monitoring glial proliferation-related substantia nigra-striatal lesions. 5. High drug doses/mice weighing less than 22 grams/mixing different batches of drug/mice not acclimatized beforehand/low animal room temperature, etc., can all lead to mouse mortality. It is recommended to increase the number of animals per group and adjust to the optimal dose according to experimental conditions. Model indicators: Substantia nigra striatum damage: After successful modeling, the activity of tyrosine hydroxylase in the substantia nigra and striatum decreased (IHC, IF, WB, etc.); Other markers: Decreased neurotransmitters in the brain (DA, DOPAC, 5-HT, HVA, etc.) (HPLC detection); Activation of microglia (IBA1+ cells) and astrocytes (GFAP+ cells) in the substantia nigra striatum, and an increase in the number of α-synuclein aggregates in the substantia nigra.

[1]. Langston J W, Irwin I. MPTP Neurotoxicity: An Overview and Characterization of Phases of Toxicity. II. Selective Accumulation of MPP in the Substantia Nigra: A Key to Neurotoxicity (Question). Life Sci., 1985, 36, No. 3, 201-12.

[2]. Potentiation of MPTP by 4-Phenylpyridine on the Neuromuscular Blockade in Mouse Phrenic Nerve-Diaphragm. Neuropharmacology, 1993, 32, No. 9, 877-83.

[3]. Gas1 up-regulation is inducible and contributes to cell apoptosis in reactive astrocytes in the substantia nigra of LPS and MPTP models. J Neuroinflammation. 2016 Jul 8;13(1):180.

[4]. Jackson-Lewis V, Przedborski S. Protocol for the MPTP mouse model of Parkinson's disease. Nat Protoc. 2007;2(1):141-51.

[5]. The CD200R1 microglial inhibitory receptor as a therapeutic target in the MPTP model of Parkinson's disease. J Neuroinflammation. 2021 Apr 6;18(1):88.

[6]. MPTP-driven NLRP3 inflammasome activation in microglia plays a central role in dopaminergic neurodegeneration. Cell Death Differ. 2019 Jan;26(2):213-228.

[7]. Reassessment of subacute MPTP-treated mice as animal model of Parkinson's disease. Acta Pharmacol Sin. 2017 Oct;38(10):1317-1328.

[8]. A sheep model for MPTP induced Parkinson-like symptoms. Life Sci. 1989;45(17):1601-8.

[9]. Langston J W, Irwin I. MPTP Neurotoxicity: An Overview and Characterization of Phases of Toxicity. II. Selective Accumulation of MPP in the Substantia Nigra: A Key to Neurotoxicity (Question). Life Sci., 1985, 36, No. 3, 201-12.

[10]. Potentiation of MPTP by 4-Phenylpyridine on the Neuromuscular Blockade in Mouse Phrenic Nerve-Diaphragm. Neuropharmacology, 1993, 32, No. 9, 877-83.

[11]. Gas1 up-regulation is inducible and contributes to cell apoptosis in reactive astrocytes in the substantia nigra of LPS and MPTP models. J Neuroinflammation. 2016 Jul 8;13(1):180.

[12]. Jackson-Lewis V, Przedborski S. Protocol for the MPTP mouse model of Parkinson's disease. Nat Protoc. 2007;2(1):141-51.

C12H15N

173.25

28289-54-5

MPTP hydrochloride

Off-white to light yellow solid

CN1CC=C(CC1)C2=CC=CC=C2

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: 本产品在运输和储存过程中需避光（避免光照）。

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

DMSO : ~50 mg/mL (~288.60 mM; with sonication and warming)

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.)