Volume 3 • Issue 4 • 1000129
J Alzheimers Dis Parkinsonism
ISSN: 2161-0460 JADP, an open access journal
Open Access Research Article
Schmitt et al., J Alzheimers Dis Parkinsonism 2013, 3:4
http://dx.doi.org/10.4172/2161-0460.1000129 Alzheimer’s Disease &
Parkinsonism
Keywords: MR-spectroscopy; C57BL/6; Brain metabolites; Absolute
quantification
Introduction
Rodent models of human diseases are of high importance
for biomedical research as they enable studying diseases under
laboratory conditions to provide information on pathophysiology of
the disease and to develop and evaluate treatment options. is is of
special importance for orphan diseases such as the guanidinoacetate
methyltransferase (GAMT) deficiency, where it has not been possible,
due to the low number of patients, to evaluate and modify treatment
strategies in a clinical setting. Further, in order to understand disease
induced changes occurring during brain development, it is essential
to have an accurate understanding of normal brain maturation and its
biological variability [1].
Proton magnetic resonance spectroscopy (
1
H MRS) is a well-
established tool for non-invasive detection and quantification of brain
metabolites such as taurine, creatine, choline, glutamate, glutamine,
myo-inositol, and N-acetylaspartate [2].
1
H MRS applied to the brain
of mice and rats at different magnetic field strengths revealed metabolic
patterns that are comparable to those observed in human brain [2-
5]. Studies of cerebral metabolite concentrations in different mouse
strains, however, have also demonstrated small (<13%) inter-strain
variations, which are limited to specific metabolites [4,6,7]. Metabolite
changes occurring during post-natal mouse brain development
have been studied in vivo with single-voxel spectroscopy [8-10] or a
*Corresponding author: Andreas Schulze, The Hospital for Sick Children, The
Research Institute, Genetics and Genome Biology, University of Toronto, 555
University Avenue, Toronto, ON, M5G 1X8, Canada, Tel: +1 (416) 813 7654; Fax: +1
(416) 813 5345; E-mail: andreas.schulze@sickkids.ca
Received August 27, 2013; Accepted November 09, 2013; Published November
13, 2013
Citation: Schmitt B, vonBoth I, Amara CE, Schulze A (2013) Quantitative
Assessment of Metabolic Changes in the Developing Brain of C57BL/6 Mice by In
Vivo Proton Magnetic Resonance Spectroscopy. J Alzheimers Dis Parkinsonism 3:
129. doi: 10.4172/2161-0460.1000129
Copyright: © 2013 Schmitt B, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Quantitative Assessment of Metabolic Changes in the Developing Brain
of C57BL/6 Mice by In Vivo Proton Magnetic Resonance Spectroscopy
Benjamin Schmitt
1
, Ingo vonBoth
2
, Catherine E Amara
3
and Andreas Schulze
4
*
1
Department of Radiology, Medical University of Vienna Centre for High-Field MR, Austria
2
Department of Laboratory Medicine and Pathobiology, University of Toronto, Canada
3
Faculty of Kinesiology & Physical Education, University of Toronto, Canada
4
The Hospital for Sick Children, The Research Institute, Genetics and Genome Biology (Work was conducted here), University of Toronto, Canada
Abstract
Localized proton MRS was used to quantify cerebral metabolite concentrations in the thalamus of mice to assess
the variation of major metabolites during brain development.
Three sets of C57BL/6 mice were followed in a longitudinal study from a very early phase at post-natal day
four (p4) until day 57 (p57). Experiments were conducted in accordance with Canadian animal care guidelines
on a 7-Tesla small animal MR system. Specimens were examined under inhalation anesthesia using single-voxel
MRS. A cubic volume with edge lengths of 1.9 mm was placed in the thalamus region of animals and point-resolved
spectroscopy (PRESS) spectra were acquired with the following parameters (TR/TE/NEX=2500 ms/20 ms/600;
Bandwidth=4000 Hz). Absolute metabolite quantification using LCModel was obtained by assigning water signal
intensity measured by MRS to water concentrations determined by histobiochemical analysis and interpolation.
Optimized anesthesia, immobilization, and careful monitoring led to a survival rate of 100% throughout the study.
The brain water content was 84.8, 78.8, and 77.6% at p12, p31, and p66. Variation of metabolites revealed similar
patterns for the total of creatine and phosphocreatine (tCr), glutamate and glutamine (Glx), and the total of N-acetyl
aspartic compounds (tNAA), with steady increases from p4 to reaching a plateau after p21. The total of Choline-
containing compounds (tCho) and myo-inositol (Ins) had high concentrations at early exam points, decreased to
minima between p14 and p19, and increased to adult levels thereafter. Taurine (Tau) had highest levels at p4,
decreased persistently but fast in the early development and slow in the later stages of brain development.
Our results indicate that biological variance must be considered if results from studies on mouse models of
pathologies are compared with results from healthy controls during development. This aspect seems to be especially
important between p10 and p21. Due to the high temporal resolution used at early time points in our study and the
inclusion of multiple groups of animals at time points, our data contribute important aspects to the existing literature
about mouse brain development.
multi-voxel approach in C57BL/6 mice, and in vitro with whole brain
samples from C57BL/6 mice [11,12]. e four in vivo studies used
different approaches for absolute metabolite quantification, namely
advanced method for accurate, robust and efficient spectral fitting
(AMARES), and linear combination of model spectra (LCModel).
Additionally, the particular time points, the intervals between them,
and the brain regions examined varied between the studies. While
the results of studies seem fairly homogeneous, distinct differences in
reported development patterns were found in a study by Larvaron et
al. [8-11]. Differences between the study results may be attributed to
the respective employed quantification strategies, e.g., the choice of
using an external quantification or measured brain water content as