Six- to eight-weeks old C57BL/6 mice were purchased from Harlan, Indianapolis, IN. Animal maintenance and experiments were in accordance with National Institutes of Health guidelines and were approved by the Institutional Animal Care and Use committee of the Rush University Medical Center, Chicago, IL. For acute MPTP intoxication, mice received four intraperitoneal (i.p.) injections of MPTP-HCl (18 mg/kg of free base; Sigma Chemical Co., St. Louis, MO) in saline at 2-h intervals (Ghosh et al. 2007 (link); Ghosh et al. 2009 (link); Roy et al. 2012 (link)). Control animals received only saline.
Mptp hcl
Manufactured by Merck Group
Sourced in United States, Italy, Sao Tome and Principe
MPTP-HCl is a chemical compound used in research applications. It is a hydrochloride salt of the compound MPTP, which is commonly used as a neurotoxin in animal studies. The primary function of MPTP-HCl is to facilitate controlled and reproducible experiments in a laboratory setting.
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Lab products found in correlation
C57bl 6j control mice, by Jackson ImmunoResearch (2 mentions)
Il 6 ko, by Jackson ImmunoResearch (2 mentions)
C57bl 6j wt, by Jackson ImmunoResearch (2 mentions)
C57bl 6j wt mice, by Jackson ImmunoResearch (2 mentions)
Epigallocatechin gallate (egcg), by Merck Group (2 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (2 mentions)
Homovanillic acid hva, by Merck Group (1 mentions)
Polyclonal rabbit anti th, by Merck Group (1 mentions)
α tubulin antibody, by Abcam (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (1 mentions)
Biotinylated secondary antibody, by Vector Laboratories (1 mentions)
C57bl j, by Charles River Laboratories (1 mentions)
Normal saline, by Merck Group (1 mentions)
Chloroquine phosphate, by MedChemExpress (1 mentions)
Parkin, by Santa Cruz Biotechnology (1 mentions)
Cathepsin d, by Santa Cruz Biotechnology (1 mentions)
α syn, by Abcam (1 mentions)
Lamp2, by Proteintech (1 mentions)
Pink1, by Santa Cruz Biotechnology (1 mentions)
Cathepsin b, by Proteintech (1 mentions)
82 protocols using mptp hcl
1
Acute MPTP Intoxication in C57BL/6 Mice
2
Primate Model of Parkinson's Disease
3
Acute MPTP Mouse Parkinson's Model
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Groups of mice (n = 5) received MPTP under an acute protocol. Mice were given 4 i.p. injections of MPTP-HCl (Sigma) 2 hours apart and at a dose of 20 mg/kg (free-base). They were euthanized 7 days after the last MPTP injection. Control mice received an equivalent volume of 0.9% NaCl solution. Removed brains were postfixed overnight in fresh 4% paraformaldehyde (PFA)/phosphate-buffered saline (PBS) solution, cryoprotected with 30% sucrose in 0.1 M PB, and frozen in isopentane (−30°C). Brain free-floating sections (20 μm thick) encompassing the entire midbrain were prepared using a freezing microtome (Microm, Germany). Three different sections, representative of three different rostrocaudal levels of the midbrain, were used. The sections were dropped with specific orientation in paraffin. The paraffin blocks were sectioned into 4 μm thick sections. All sections were mounted on silan-coated glass slides and used for morphological, immunohistochemical, and immunofluorescence analysis. Samples of three different sections (20 μm thick) were collected and used for immunoblotting analysis.
4
Acute MPTP Intoxication in C57BL/6 Mice
5
MPTP-Induced Parkinson's Model in Mice
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C57BL/6 mice aged 8–10 weeks were kept individually in a cage with free access to water and food and lived in a 12 h light/12 h darkness cycle. Animal care and the general protocols for animal use and MPTP experiments were approved by the Institutional Animal Care and Use Committee of Chang Gung Memorial Hospital (IACUC Approval No: 2017031401). There were four treatment groups of animals, including a sham control group (I), an MPTP group (II), an erinacine A group (III, 1 mg/kg,) and two H. erinaceus wet mycelia (HEM) groups (III, 10.76 mg and IV, 21.52 mg). Accordingly, the mice were intraperitoneally (i.p.) injected with MPTP-HCl (30 mg/kg; Sigma, St. Louis, MO) (the MPTP group) or saline (the control group) over 4 days. After the first MPTP injection, the mice received HEM (dissolved in H2O; HEM groups) with oral administration or erinacine A (dissolved in dimethyl sulfoxide (DMSO); erinacine A groups) with intraperitoneal administration or an equivalent volume of saline (sham-operated group) for an additional 5 days. The mice were sacrificed 8 days after MPTP injection and their brains were collected for further analysis [25 (link)].
6
MPTP-induced Parkinson's Disease Mouse Model
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All animal experiments involved in this research were conducted under the approval of animal ethics committee of The Affiliated Brain Hospital of Nanjing Medical University.
C57BL/6 mice (male, 10 weeks old, 20-25 g, n = 30) were provided from the Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (Beijing, China). Mice were kept in individual cages in a 12h light/dark cycle constant temperature room (25° C) with free access to food and water.
MPTP-HCl (Sigma-Aldrich, St. Louis, MO, USA) was used to construct PD mouse model. Briefly, mice (n = 24) were injected intraperitoneally with MPTP-HCl with a dose of 30 mg/kg/day for 4 consecutive days. The other 6 mice were served as Sham group and they were injected intraperitoneally with 0.9% sterile saline with an equivalent volume for 4 consecutive days. On the 1st, 3rd, 5th and 7th day after the last injection of MPTP-HCl, 6 mice were randomly selected and sacrificed to collect the midbrain. The midbrain was immediately stored at -80° C.
C57BL/6 mice (male, 10 weeks old, 20-25 g, n = 30) were provided from the Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (Beijing, China). Mice were kept in individual cages in a 12h light/dark cycle constant temperature room (25° C) with free access to food and water.
MPTP-HCl (Sigma-Aldrich, St. Louis, MO, USA) was used to construct PD mouse model. Briefly, mice (n = 24) were injected intraperitoneally with MPTP-HCl with a dose of 30 mg/kg/day for 4 consecutive days. The other 6 mice were served as Sham group and they were injected intraperitoneally with 0.9% sterile saline with an equivalent volume for 4 consecutive days. On the 1st, 3rd, 5th and 7th day after the last injection of MPTP-HCl, 6 mice were randomly selected and sacrificed to collect the midbrain. The midbrain was immediately stored at -80° C.
7
Immunohistochemistry and HPLC Analysis of Dopaminergic Markers
8
MPTP-Induced Parkinson's Model in TREM2 Knockout Mice
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TREM2 -/-mice were generated as described (Turnbull, et al., 2006) . TREM2 -/-mice and C57BL/6 control littermates (8-12 weeks of age) received 4 intra-peritoneal (i.p.) injections at 2 hours intervals of either vehicle (PBS) or MPTP-HCl (20 mg/kg of free base in PBS; Sigma-Aldrich, Italy). This dose was chosen as it has been shown to be the lowest producing significant loss of striatal dopamine and dopamine active transporter (DAT) levels with a concomitant increase in striatal and nigral activated microglia expressing TSPO (Jackson-Lewis and Przedborski, 2007) (link). MPTP handling and safety measures were in accordance with published guidelines. Radiopharmaceuticals used for ex vivo and in vivo studies were prepared in our facility as described below. Animal experiments were carried out in compliance with the institutional guidelines for the care and use of experimental animals (IACUC), which have been notified to the Italian Ministry of Health and approved by the Ethics Committee of the San Raffaele Scientific Institute.
9
MPTP-Induced Parkinson's Model in Rhesus Monkeys
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Example 2
Female rhesus monkeys (6-8 years old; 5-7 kg) were used in this study. All animals were singly housed with a 12-h light/dark cycle. Purine monkey chow and water were available ad libitum. Diets were supplemented with fruit during the testing sessions. The study was performed in accordance with federal guidelines of proper animal care and with the approval of the IACUC. Monkeys were intoxicated with MPTP according to protocol described previously (12-16). Briefly, animals were tranquilized with ketamine (10 mg/kg, i.m.) and then maintained on an anesthetic plane with isoflurane (1-2%). The animals were put in the supine position. For each injection, a right-sided incision was made along the medial edge of the sternocleidomastoid muscle. The carotid sheath was opened and the common carotid artery, internal jugular vein, and vagus nerves were identified. The common carotid was exposed below the carotid bifurcation. The external carotid artery was then ligated. A 27 gauge butterfly needle was inserted into the common carotid artery in a direction retrograde to blood flow; for each injection, 20 ml of saline containing 3 mg of MPTP-HCl (Sigma) was infused at a rate of 1.33 ml/min (15 min). After the infusion was completed, 3 ml of saline was delivered, and then the incision was closed.
10
Murine models of neurodegeneration
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C57BL6/J WT (Jackson Labs, 000664), IL-6 KO (Jackson Labs, 002650) mice were obtained from the Jackson Laboratories. GFAP-IL6 transgenic mice were obtained from Scripps Research Institute via material transfer agreement (Castelnau et al., 1998 (link)). For toxin-induced dopaminergic neurodegeneration, 12-week-old C57BL6/J WT mice from Jackson Labs (000664) were given a single subcutaneous injection of either 20 mg/kg MPTP-HCl (Sigma Aldrich, M0896) or normal saline. PBS-and α-syn PFF-injected mice were treated with 0.5 mg/mL deferiprone (ApoPharma) dissolved in their drinking water starting 1 week after injection and continuing until euthanasia. Previous data have shown that this dose is sufficient to reduce iron-induced neurodegeneration in mice (Zhao et al., 2015 (link)). Hemizygous hA53T transgenic mice (006823) were obtained from Jackson Laboratories, euthanized at 12 months of age and compared to age- and sex-matched C57BL6/J control mice (Jackson Labs, 000664). All housing, breeding, and procedures were performed according to the NIH Guide for the Care and Use of Experimental Animals, ARVO standards for the use of animals and approved by either the Johns Hopkins University Animal Care and Use Committee or the University of Pennsylvania Animal Care and Use Committee.
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