Mouse MPTP Model
Description:
Parkinson’s Disease Mouse Model. Twelve, 14 weeks old male C57BL/6J mice (Jackson labs) were used for the study. Half were treated with MPTP (n=6) and the other half (n=6) were control mice, treated with saline. The mice were given intraperitioneal (i.p.) injections of 30mg/kg MPTP free base (MPTP.Hcl, Sigma) once a day for five days. They were sacrificed four days after the last treatment. All studies were performed in accordance with the UCLA Animal Research Committee guidelines and approval.
Dissecting Regions of Interest. Frontal cortex, midbrain and striatum were dissected on a cold plate. A Rodent Brain Matrix was used to create one mm coronal slices from which the region of interest were cut out using online resources from the Allen Brain Atlas (www.brainatlas.org) and Mouse Brain Library (www.mbl.org) as reference guides. The brain region identified as Frontal Cortex in this work corresponds to the dissection of the cortical area in each 1 mm slice between Bregma coordinates +3 and 0. Striatum was dissected between Bregma coordinates +1 and -2. Midbrain was dissected between Bregma coordinates -3 and -5 by removing Thalamus, Hypothalamus and Pons.
Total RNA Isolation and Quality Check. Total RNA was isolated from the dissected brain samples using the RNeasy Lipid Tissue Mini Kit (Qiagen, USA). Tissue lysis and homogenization was done with QIAzol Lysis Reagent and a Tissue Tearor rotor stator (Biospec products, USA). Total RNA quality was assessed by microcapillary electrophoresis using the BioAnalyzer 2100 (Agilent, USA). All samples were of excellent quality (RIN > 8).
DNA Microarray. A total of 18 samples of total RNA from the control and MPTP treated mice were processed by the UCLA Microarray Core Facility and hybridized to the microarrays following the manufacturer protocol. We used the Affymetrix GeneChip Mouse Genome 430 2.0 array, which contains the entire mouse genome on one chip (~45K features).
Microarray Data Analysis. Absolute gene expressions were calculated using Robust Microarray Analysis (RMA) (Irizarry et al., 2003). In the RMA method, Perfect Match intensities are background corrected and normalized using quantile normalization (all arrays are normalized at the same time) (Bolstad et al., 2003).
Bolstad, B.M., R. A. Irizarry, M. Astrand and T. P. Speed (2003). "A comparison of normalization methods for high density oligonucleotide array data based on variance and bias." Bioinformatics 19: 185-193.
Irizarry, R. A., B. Hobbs, F. Collin, Y. D. Beazer-Barclay, K. J. Antonellis, U. Scherf and T. P. Speed (2003). "Exploration, normalization, and summaries of high density oligonucleotide array probe level data." Biostatistics 4: 249-264.
Usage: This dataset can be viewed using the Mouse BIRN Atlasing Tool (MBAT). See the respective manual for the program.
Accession Number: TBD
Contributors/Authors:
Daniel M. Sforza 1 , Parvana Hartenstein 1, Goran Lacan 2, William P. Melega 2, and Arthur W. Toga 1
1. UCLA Laboratory of Neuro Imaging, Department of Neurology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095-7334, USA.
2. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
Citations:
- D. M. Sforza, P. Hartenstein, G. Lacan, W. P. Melega, and A. W. Toga. Mapping of multiple brain region gene expression in a mouse MPTP model of Parkinson's disease. Presented at Neuroscience 2006, meeting of the Society for Neuroscience, Atlanta, October 14-18, 2006.
- Please acknowledge the contributors and the mouse BIRN.
Technical Contacts:
Daniel Sforza sforza@ucla.edu
Acknowledgements: This work was made possible by Grant Number U24 RR021760 from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NCRR or NIH.