Imaging with time-of-flight secondary ion mass spectrometry (TOF-SIMS) has expanded very rapidly with the development of gold cluster ion sources (Au 3 ؉ ). It is now possible to acquire ion density maps (ion images) on a tissue section without any treatment and with a lateral resolution of few micrometers. In this article, we have taken advantage of this technique to study the degeneration/regeneration process in muscles of a Duchenne muscular dystrophy model mouse. Specific distribution of different lipid classes (fatty acids, triglycerides, phospholipids, tocopherol, coenzyme Q9, and cholesterol) allows us to distinguish three different regions on a mouse leg section: one is destroyed, another is degenerating (oxidative stress and deregulation of the phosphoinositol cycle), and the last one is stable. TOF-SIMS imaging shows the ability to localize directly on a tissue section a great number of lipid compounds that reflect the state of the cellular metabolism. In lipidomics, as in all biological domains, the simultaneous localization of numerous organic compounds in tissue sections is a crucial issue for understanding cellular metabolic processes. New tools are developed to acquire the maximum amount of data in a short time, and the use of mass spectrometry images to manage information on biological tissue surfaces is increasing rapidly.Two mass spectrometry imaging methods have been developed. The first one is based on matrix-assisted laser desorption ionization (MALDI) (1, 2) coupled with a timeof-flight (TOF) technique, which leads to the acquisition of images in a mass range from 500 to a few tens of thousands daltons with a spatial resolution of ف 50 m (3, 4), and the second one is based on TOF secondary ion mass spectrometry (SIMS), which leads to the acquisition of images with masses of Ͻ 1,000 Da with a spatial resolution of 1 m or less (5-7). In both cases, tissue sections having a thickness of 10-20 m are cut using a cryostat at Ϫ 20 Њ C and transferred to target plates (stainless steel or glass). For MALDI-TOF experiments, the matrix is deposited by air spray, whereas no preliminary sample coating is needed for TOF-SIMS experiments. A laser (MALDI) or a focused ion beam (SIMS), both pulsed, irradiates the sample at each surface element, which is called a spot, and from which a mass spectrum is acquired and recorded. Each spot corresponds to a pixel of the image, and the intervals between each spot define the lateral resolution. The inevitable use of a matrix could be a weakness of the MALDI technique: homogeneous deposition by air spray is difficult to control, and the signal visibility is low for m/z Ͻ 600 because of mass interference with matrix ion peaks (8). Tandem mass spectrometry imaging experiments can extend the accessible mass range down to m/z ف 100-200, but to the detriment of the spatial resolution, which is only 500 m with present tandem mass spectrometers (9). Future improvements are expected toward a better spatial resolution of 1-10 m in association with reasonable data...
Duchenne muscular dystrophy (DMD) is a lethal, Xlinked disorder associated with dystrophin deficiency that results in chronic inflammation, sarcolemma damage, and severe skeletal muscle degeneration. Recently, the use of L-arginine , the substrate of nitric oxide synthase (nNOS), has been proposed as a pharmacological treatment to attenuate the dystrophic pattern of DMD. However, little is known about signaling events that occur in dystrophic muscle with L-arginine treatment. Considering the implication of inflammation in dystrophic processes, we asked whether L-arginine inhibits inflammatory signaling cascades. We demonstrate that L-arginine decreases inflammation and enhances muscle regeneration in the mdx mouse model. Classic stimulatory signals, such as proinflammatory cytokines interleukin-1, interleukin-6, and tumor necrosis factor-␣, are significantly decreased in mdx mouse muscle, resulting in lower nuclear factor (NF)-B levels and activity. NF-B serves as a pivotal transcription factor with multiple levels of regulation; previous studies have shown perturbation of NF-B signaling in both mdx and DMD muscle. Moreover, L-arginine decreases the activity of metalloproteinase (MMP)-2 and MMP-9, which are transcriptionally activated by NF-B. We show that the inhibitory effect of L-arginine on the NF-B/MMP cascade reduces -dystroglycan cleavage and translocates utrophin and nNOS throughout the sarcolemma. Collectively, our results clarify the molecular events by which L-arginine promotes muscle membrane integrity in dystrophic muscle and suggest that NF-B-related signaling cascades could be potential therapeutic targets for DMD management. (Am J Pathol
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