Electron microscopy in the investigation and diagnosis of muscle disease
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1 Electron microscopy in the investigation and diagnosis of muscle disease Roy Weller Clinical Neurosciences University of Southampton School of Medicine
2 Normal Muscle
3 Normal Muscle
4 The Sarcomere The names for the different parts are derived from German
5 The Sarcomere H-zone (from the German heller", brighter) Inside the H-zone is a thin M-line (from the German Mittelscheibe, the disc in the middle of the sarcomere) formed of cross-connecting elements of the cytoskeleton I-band = Isotropic in polarised light as the actin filaments are thin and do not interfere with the light A-band = Anisotropic in polarised light as the myosin filaments are thicker and do interfere with the light Z-line = Zwischenscheibe the disc in between the I bands
6 TS through an A-band
7
8
9 Sarcomeres
10 T-tubes and Lateral Sacs
11 Myofibils Nucleus EM of muscle fiber Sarcolemma Basal lamina or Basement membrane Pathology and Genetics of Muscle Disease Book to be published 2013 Satellite cell Macrophages Perivascular Lymphocytes Regenerating myoblast
12 Neurogenic muscle disease
13 Neurogenic Muscle Disease ATPase stain Normal Chequer-board Pattern Denervation small group atrophy Reinnervation and fibre type grouping
14 Sarcolemmal disorders including Duchenne musclar dystrophy
15 Muscular Dystrophies associated with the lack of Sarcolemmal proteins Basement Membrane
16 Duchenne Muscular Dystrophy Dystrophin Immunocytochemistry- Normal
17 Normal Duchenne Muscular dystrophy: separation of basement membrane from the muscle fibre
18 Duchenne Muscular dystrophy: separation of basement membrane from the muscle fibre
19 Ion Channel disorders
20 Nerve and muscle end plate Cholinesterase in the Basement Membrane Sarcolemma Genetic Ca Disorders of Sarcoplasmic reticulum 2+ Ion Channels Mitochondria Glycogen and lipid ATP Ca 2+ T-tube Release and uptake of Ca2+ Muscle fibril contraction and relaxation
21 T-tubules and Lateral Sacs
22 D Myopathy with Tubular Aggregates
23 Tubular Aggregates
24 Tubular Aggregates
25 Tubular Aggregates
26 Mitochondrial myopathies
27 Mitochondrial Cytopathy (Myopathy)
28 Mitochondrial myopathy 1 μm resin section
29 Mitochondrial Cytopathy (Myopathy) (megaconial)
30 Inclusions in Mitochondrial Cytopathy (Myopathy)
31 Inclusions in Mitochondrial Cytopathy (Myopathy)
32 Cytoplasmic Inclusions
33 Cytoplasmic Inclusions
34 Cytoplasmic Inclusions some myofibrillar myopathies
35 Cytoplasmic Inclusion
36 Sarcoglycan & Dystroglycan Complexes Laminin 2 Caveolin Basement Membrane Sarcolemma Dystrophin F-Actin Z-band A-Band Plectin Desmin etc Actin Myosin Z-bands Desmin & intermediate filamentassociated proteins αb-crystallin Desmin etc Myofibrillar myopathies Emerin Lamin A&C Nucleus
37 Reducing body reduces menadione nitro-blue-tetrazolium (NBT) mutation in the FHL1 gene and other disorders
38 Z-band disorders rod bodies
39 Nemaline myopathy defects in thin filament proteins eg actin
40 Toxic myopathy due to Senna
41 Muscle fibre destruction and regeneration For example inflammatory myopathies Polymyositis, dermatomyositis, Inclusion Body Myositis, parasitic infections But also Dystrophies
42 Inflammatory cells in the endomysium surrounding atrophic muscle fibres
43 Hunkin Muscle fibre destruction influenza-a myopathy macrophages and regenerating myoblasts
44 Normal Necrosis 24hrs Regeneration 10-14days 30days 30days Focal necrosis and regeneration of skeletal muscle
45 Satellite Cell source of regenerating myoblasts
46 Normal basement membrane (Laminin) Trypsin destroys the basement membrane Barium chloride destroys only the muscle fibres Is an intact basement membrane tube essential for successful muscle fibre regeneration? Role of the basement membrane in the regeneration of skeletal muscle. Caldwell CJ, Mattey DL, Weller RO Neuropathol Appl Neurobiol Jun;16:
47 Scanning Electron microscopy Barium chloride destroys only the muscle fibres Basement membrane tubes are preserved
48 Scanning Electron microscopy: Macrophages on basement membrane tubes: -- Barium chloride treated muscle
49 * Scanning Electron microscopy: Macrophages within basement membrane tube: - - Barium chloride treated muscle
50 Macrophages and regenerating myoblast within basement membrane tube: -- Barium chloride treated muscle
51 Basement membrane closely invests a regenerating myoblast in Trypsin treated muscle in which the muscle fibre and the basement membrane were destroyed
52 5 days recovery: which is the Trypsin-treated muscle?
53 Inclusion Body Myositis
54 Inclusion Body Myositis
55 Inclusion Body Myositis
56 Inclusion Body Myositis
57 Inclusion Body Myositis
58 Inclusion Body Myositis
59 Inclusion Body Myositis
60 Electron microscopy in the evaluation of individual cases Dr Ingrid Mazanti Dr Mark Walker
61
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