This study compared the metabolic and activation changes induced by electrically-evoked (NMES) and voluntary (VOL) contractions performed at the same submaximal intensity using P chemical shift imaging (CSI) and T2 mapping investigations.
Neuromuscular electrostimulation (NMES) leads to a spatially fixed, synchronous and superficial motor unit recruitment which could induce muscle damage. Therefore, the extent of muscle damage and its spatial occurrence was expected to be heterogeneous across and along quadriceps femoris (QF) muscles. The aim of the present study was to characterize muscle spatial heterogeneity in QF damage after a single bout of isometric NMES by using multimodal magnetic resonance imaging (MRI).
We have designed and constructed an experimental set-up allowing electrical stimulation of hindlimb mouse muscles and the corresponding force measurements at high-field (11.75T). We performed high-resolution multimodal MRI (including T2 -weighted imaging, angiography and diffusion) and analysed the corresponding MRI changes in response to a stimulation protocol. Mice were tested twice over a 1-week period to investigate the reliability of mechanical measurements and T2 changes associated with the stimulation protocol. Additionally, angiographic images were obtained before and immediately after the stimulation protocol. Finally, multislice diffusion imaging was performed before, during and immediately after the stimulation session. Apparent diffusion coefficient (ADC) maps were calculated on the basis of diffusion weighted images (DWI). Both force production and T2 values were highly reproducible as illustrated by the low coefficient of variation (<8%) and high intraclass correlation coefficient (?0.75) values. Maximum intensity projection angiographic images clearly showed a strong vascular effect resulting from the stimulation protocol. Although a motion sensitive imaging sequence was used (echo planar imaging) and in spite of the strong muscle contractions, motion artifacts were minimal for DWI recorded under exercising conditions, thereby underlining the robustness of the measurements. Mean ADC values increased under exercising conditions and were higher during the recovery period as compared with the corresponding control values. The proposed experimental approach demonstrates accurate high-field multimodal MRI muscle investigations at a preclinical level which is of interest for monitoring the severity and/or the progression of neuromuscular diseases but also for assessing the efficacy of potential therapeutic interventions.
Recently a new MR endogenous contrast mechanism was reported. It allows specifically imaging the magnetization transfer (MT) effect arising from inhomogeneously broadened components of the NMR spectrum, and was hence dubbed ihMT. Such unique NMR lineshape properties are presumably occurring in myelin because of its specifically ordered, multilayered sheath structure. Here, optimization of a pulsed ihMT preparation module is presented to provide guidance for future studies and improve the understanding of underlying contrast mechanisms.
Isometric contractions induced by neuromuscular electrostimulation (NMES) have been shown to result in a prolonged force decrease but the time course of the potential central and peripheral factors have never been investigated. This study examined the specific time course of central and peripheral factors after isometric NMES-induced muscle damage. Twenty-five young healthy men were subjected to an NMES exercise consisting of 40 contractions for both legs. Changes in maximal voluntary contraction force of the knee extensors (MVC), peak evoked force during double stimulations at 10 Hz (Db(10)) and 100 Hz (Db(100)), its ratio (10:100), voluntary activation, muscle soreness and plasma creatine kinase activity were assessed before, immediately after and throughout four days after NMES session. Changes in knee extensors volume and T2 relaxation time were also assessed at two (D2) and four (D4) days post-exercise. MVC decreased by 29% immediately after NMES session and was still 19% lower than the baseline value at D4. The decrease in Db(10) was higher than in Db(100) immediately and one day post-exercise resulting in a decrease (-12%) in the 10:100 ratio. On the contrary, voluntary activation significantly decreased at D2 (-5%) and was still depressed at D4 (-5%). Muscle soreness and plasma creatine kinase activity increased after NMES and peaked at D2 and D4, respectively. T2 was also increased at D2 (6%) and D4 (9%). Additionally, changes in MVC and peripheral factors (e.g., Db(100)) were correlated on the full recovery period, while a significant correlation was found between changes in MVC and VA only from D2 to D4. The decrease in MVC recorded immediately after the NMES session was mainly due to peripheral changes while both central and peripheral contributions were involved in the prolonged force reduction. Interestingly, the chronological events differ from what has been reported so far for voluntary exercise-induced muscle damage.
Nemaline myopathy (NM), the most common non-dystrophic congenital disease of skeletal muscle, can be caused by mutations in the skeletal muscle ?-actin gene (ACTA1) (~25% of all NM cases and up to 50% of severe forms of NM). Muscle function of the recently generated transgenic mouse model carrying the human Asp286Gly mutation in the ACTA1 gene (Tg(ACTA1)(Asp286Gly)) has been mainly investigated in vitro. Therefore, we aimed at providing a comprehensive picture of the in vivo hindlimb muscle function of Tg(ACTA1)(Asp286Gly) mice by combining strictly noninvasive investigations. Skeletal muscle anatomy (hindlimb muscles, intramuscular fat volumes) and microstructure were studied using multimodal magnetic resonance imaging (Dixon, T2, Diffusion Tensor Imaging [DTI]). Energy metabolism was studied using 31-phosphorus Magnetic Resonance Spectroscopy ((31)P-MRS). Skeletal muscle contractile performance was investigated while applying a force-frequency protocol (1-150 Hz) and a fatigue protocol (6 min-1.7 Hz). Tg(ACTA1)(Asp286Gly) mice showed a mild muscle weakness as illustrated by the reduction of both absolute (30%) and specific (15%) maximal force production. Dixon MRI did not show discernable fatty infiltration in Tg(ACTA1)(Asp286Gly) mice indicating that this mouse model does not reproduce human MRI findings. Increased T2 values were observed in Tg(ACTA1)(Asp286Gly) mice and might reflect the occurrence of muscle degeneration/regeneration process. Interestingly, T2 values were linearly related to muscle weakness. DTI experiments indicated lower ?2 and ?3 values in Tg(ACTA1)(Asp286Gly) mice, which might be associated to muscle atrophy and/or the presence of histological anomalies. Finally (31)P-MRS investigations illustrated an increased anaerobic energy cost of contraction in Tg(ACTA1)(Asp286Gly) mice, which might be ascribed to contractile and non-contractile processes. Overall, we provide a unique set of information about the anatomic, metabolic and functional consequences of the Asp286Gly mutation that might be considered as relevant biomarkers for monitoring the severity and/or the progression of NM and for assessing the efficacy of potential therapeutic interventions.
Different MR techniques, such as relaxation times, diffusion, perfusion, and spectroscopy have been employed to study rodent spinal cord. In this chapter, a description of these methods is given, along with examples of normal metrics that can be derived from the MR acquisitions, as well as examples of applications to pathology.
Perfusion MRI is a tool to assess the spatial distribution of microvascular blood flow. Arterial spin labeling (ASL) is shown here to be advantageous for quantification of cerebral microvascular blood flow (CBF) in rodents. This technique is today ready for assessment of a variety of murine models of human pathology including those associated with diffuse microvascular dysfunction. This chapter provides an introduction to the principles of CBF measurements by MRI along with a short overview over applications in which these measurements were found useful. The basics of commonly employed specific arterial spin-labeling techniques are described and theory is outlined in order to give the reader the ability to set up adequate post-processing tools. Three typical MR protocols for pulsed ASL on two different MRI systems are described in detail along with all necessary sequence parameters and technical requirements. The importance of the different parameters entering theory is discussed. Particular steps for animal preparation and maintenance during the experiment are given, since CBF regulation is sensitive to a number of experimental physiological parameters and influenced mainly by anesthesia and body temperature.
With the increasing number of transgenic mouse models of human brain diseases, there is a need for a sensitive method that allows assessing quantitative whole brain perfusion within a reasonable scan time. Arterial spin labeling (ASL), an MRI technique that permits the noninvasive quantification of cerebral blood flow, has been used to assess rodents brain perfusion. For mice, the reported experiments performed with continuous or pulsed ASL were challenged by poor multislice capability, limited sensitivity, or quantification issues. Here, the recently proposed pseudo-continuous ASL strategy, which has shown great promise for human studies, was investigated for mouse brain perfusion imaging at 11.75 T. Pseudo-continuous ASL was experimentally optimized and compared with a standard flow-sensitive alternating inversion recovery sequence for sensitivity, robustness, absolute quantification, and multislice imaging capability. A sensitivity gain up to 40% and clear advantages for multislice imaging are obtained with pseudo-continuous ASL.
Diffusion tensor imaging is increasingly used for probing spinal cord (SC) pathologies, especially in mouse models of human diseases. However, diffusion tensor imaging series requires a long acquisition time and mouse experiments rarely use rapid imaging techniques such as echo planar imaging. A recent preliminary study demonstrated the feasibility and robustness of the echo planar imaging sequence for mouse cervical SC diffusion tensor imaging investigations. The feasibility of echo planar imaging at thoracic and lumbar levels, however, remained unknown due to bulk motion, field inhomogeneities, and off-centering of the SC in the axial plane. In the present study, the feasibility and the robustness of an echo planar imaging-based diffusion tensor imaging sequence for mouse thoracic and lumbar SC investigations is demonstrated. Quantitative and accurate diffusion tensor imaging metrics, as well as high spatially resolved images, have been obtained. This successful demonstration may open new perspectives in the field of mouse SC imaging. Echo planar imaging is used in several imaging modalities, such as relaxometry or perfusion, and may prove to be very attractive for multimodal MR investigations to acquire a more detailed characterization of the SC tissue.
In spinal cord injuries (SCI), tissue edema and consequent ischemia play an important role in neuronal damage. The assessment of quantitative spinal cord blood flow (SCBF) would be very valuable to help in understanding SCI pathophysiology. SCBF has previously been measured in animals with invasive techniques such as hydrogen clearance or labeled microspheres. A recent preliminary study also demonstrated the feasibility of assessing cervical SCBF by MRI with arterial spin labeling (ASL). However, due to bulk motion and field inhomogeneities, the feasibility of perfusion MRI at lower levels of the SC (thoracic, lumbar) remained an open question. In the present study, absolute SCBF measurements were carried out at both the cervical C3 and lumbar L1 levels of mouse SC using an adapted presaturated flow-sensitive alternating inversion recovery (presat-FAIR) ASL technique at 11.75T. Quantitative SCBF maps (resolution of 133 x 133 microm(2)) showed significantly lower gray matter (GM) perfusion values at the L1 level as compared to the C3 level (6% and 11% for the ventral and dorsal horns and 8% for total GM). The presat-FAIR technique was then successfully applied to a mouse model of hemisection performed at the L1 level, illustrating the potential of ASL to help in SC pathology characterization.
MR spectroscopy allows a noninvasive assessment of metabolic information in healthy and pathological central nervous system. Whereas MR spectroscopy has been extensively applied in the brain, only few spectroscopic studies of the spinal cord (SC) have been performed so far. For mice, due to additional technical challenges, in vivo 1H SC MRS has not yet been reported. In this work, the feasibility of short echo time localized proton magnetic resonance spectroscopy using Point RESolved Spectroscopy sequence for the examination of mouse cervical SC at 11.75 T is presented. Several optimizations were performed to improve the static field homogeneity, to reduce physiological motion effects and lipid contaminations arising from SC surrounding tissues, and to provide a careful metabolic quantification. Satisfactory spectrum quality was obtained. The described protocol allowed reliable quantification of five metabolites in the cervical SC. The mean reproducibility regarding the quantification of tNAA, tCr and tCho was ?80%, >70% for mI and >55% for Glu, whereas the intersubject variabilities were ?21%. The application of this protocol to transgenic mouse models in pathological conditions such as SC injury or neurodegenerative diseases may thus provide complementary information to MRI and increase our understanding of such pathologies.
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