The Imaging Core will provide both fluorescence light microscopy and electron microscopy resources to Center Investigators to enable evaluation of muscle morphology and structural organization, and to localize specific molecular components in the sarcomeres, cytoskeleton, subcellular organelles and membranes. The broad long-term objective of this Core is to provide quantitative structural information that will be an essential link in the proposed Center's long-term goal to achieve a comprehensive understanding of multi-scale structure-function relationships in skeletal muscle. The Imaging Core will interface with the Phenotyping Core and the High-throughput Cell Analysis Core by assisting investigators in fluorescence light microscopy and transmission electron microscopy analyses of skeletal muscle tissues and cells isolated from wild-type and transgenic mice, or from healthy and diseased human muscles. The Core will provide Center investigators with training and assistance in the complex imaging technologies of Transmission Electron Microscopy and Confocal Laser Scanning Fluorescence Microscopy on fixed specimens, Confocal Spinning Disc and Wide-Field Fluorescence Microscopy of molecular dynamics in living cells, and Single-Molecule Fluorescence imaging using Total Internal Reflection Fluorescence (TIRF) Microscopy. The Core will provide training and assistance with routine aspects of sample preparation for microscopy, and training and access to microscopes in The Scripps Research Institute (TSRI) Microscopy Facility, and in the Center for Integrated Molecular Biosciences (CIMBio) Fluorescence Microscopy Suite at TSRI. The Core will also assist investigators in application of image analysis software to their experimental problems and in quantitative interpretation of fluorescent image data.
The Specific Aims are: 1) To provide training and technical assistance in skeletal muscle cell and tissue fixation and processing in preparation for fluorescence light microscopy and/or transmission electron microscopy;2) To provide training and access to fluorescence and electron microscopes at TSRI;3) To provide training and assistance in quantitative 2D and 3D image analysis software packages (Metamorph, Volocity Suite), and custom computational approaches to measure myofibril structure (Distributed Deconvolution);4) To upgrade the TIRF microscope to perform superresolution Photoactivation Localization Microscopy (PALM) and Stochastic Optical Resconstruction Microscopy (STORM), enabling nano-scale single-molecule fluorescence imaging in muscle cells and isolated myofibrils.

Public Health Relevance

Muscle contraction relies on movements of repeating structures called sarcomeres, which are attached to the muscle membranes and tendons to generate force. Muscle diseases, such as myopathies and dystrophies exhibit disruptions in sarcomeres and membrane attachments, as well as alterations in muscle fiber structure due to degeneration and regeneration. Microscopic visualization of muscle structures will explain altered function, provide insight into causes of human myopathies and lead to better treatments.

National Institute of Health (NIH)
Center Core Grants (P30)
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Special Emphasis Panel (ZAR1)
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University of California San Diego
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Chapman, Mark A; Pichika, Rajeswari; Lieber, Richard L (2015) Collagen crosslinking does not dictate stiffness in a transgenic mouse model of skeletal muscle fibrosis. J Biomech 48:375-8
Gokhin, David S; Tierney, Matthew T; Sui, Zhenhua et al. (2014) Calpain-mediated proteolysis of tropomodulin isoforms leads to thin filament elongation in dystrophic skeletal muscle. Mol Biol Cell 25:852-65
Dayanidhi, Sudarshan; Lieber, Richard L (2014) Skeletal muscle satellite cells: mediators of muscle growth during development and implications for developmental disorders. Muscle Nerve 50:723-32
Tierney, Matthew Timothy; Aydogdu, Tufan; Sala, David et al. (2014) STAT3 signaling controls satellite cell expansion and skeletal muscle repair. Nat Med 20:1182-6
Chakkalakal, Joe V; Christensen, Josef; Xiang, Wanyi et al. (2014) Early forming label-retaining muscle stem cells require p27kip1 for maintenance of the primitive state. Development 141:1649-59
Tuttle, Lori J; Alperin, Marianna; Lieber, Richard L (2014) Post-mortem timing of skeletal muscle biochemical and mechanical degradation. J Biomech 47:1506-9
Wahlquist, Christine; Jeong, Dongtak; Rojas-Muñoz, Agustin et al. (2014) Inhibition of miR-25 improves cardiac contractility in the failing heart. Nature 508:531-5
Chapman, Mark A; Zhang, Jianlin; Banerjee, Indroneal et al. (2014) Disruption of both nesprin 1 and desmin results in nuclear anchorage defects and fibrosis in skeletal muscle. Hum Mol Genet 23:5879-92
Albini, Sonia; Puri, Pier Lorenzo (2014) Generation of myospheres from hESCs by epigenetic reprogramming. J Vis Exp :e51243
Saccone, Valentina; Consalvi, Silvia; Giordani, Lorenzo et al. (2014) HDAC-regulated myomiRs control BAF60 variant exchange and direct the functional phenotype of fibro-adipogenic progenitors in dystrophic muscles. Genes Dev 28:841-57

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