Storage‐associated artefact in equine muscle biopsy samples

Stanley, Rachael; Maile, C.; Piercy, Richard J.

doi:10.2746/042516408x330374

Cited by 9 publications

(7 citation statements)

References 14 publications

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“…For multiple fluorescent labelling (Fig 1), sections were initially fixed with methanol‐acetone (1:1) at ‐20°C for 15 min and rinsed with 3 changes of phosphate‐buffered saline (PBS) over 5 min (all subsequent labelling was performed in a darkened humidified chamber at room temperature with the same rinsing protocol). First, a goat polyclonal anti‐collagen V IgG antibody 3 (1:10 dilution) was applied for 1 h followed by rinsing in PBS (Stanley et al . 2009).…”

Section: Methodsmentioning

confidence: 99%

“…For multiple fluorescent labelling (Fig 1), sections were initially fixed with methanol-acetone (1:1) at -20°C for 15 min and rinsed with 3 changes of phosphate-buffered saline (PBS) over 5 min (all subsequent labelling was performed in a darkened humidified chamber at room temperature with the same rinsing protocol). First, a goat polyclonal anti-collagen V IgG antibody 3 (1:10 dilution) was applied for 1 h followed by rinsing in PBS (Stanley et al 2009). Three separate mouse monoclonal antibodies that detect type 1 (Slow myosin IgG antibody, MAB1628) 4 , type 2a (Type 2a IgG antibody, A4.74) 6 and both type 2a and 2x (MHCf lgG antibody) 7 (Rivero et al 1996a) were individually labelled respectively, with Alexa fluor-conjugated IgG1 Fab fragments designed for 3 different emitting wavelengths (350 nm (Zenon Alexafluor 350 mouse IgG1 labelling kit 5 , 488 nm; (Zenon Alexafluor 488 mouse IgG1 labelling kit 5 and 594 nm) [(Zenon Alexafluor 594 mouse IgG1 labelling kit) 5 ] over 5 min followed by a blocking step for 5 min at room temperature according to the manufacturers' recommendations ( Table 1).…”

Section: Methodsmentioning

confidence: 99%

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Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection

Tulloch

Perkins

Piercy³

2011

Equine Veterinary Journal

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Skeletal muscle is composed of a heterogeneous mixture of several fibre types, each with specific physiological properties. In equine muscle, identification of these individual fibres (fibre typing) is important for both exercise physiology and pathological studies. Traditionally, fibre typing has been achieved by adenosine triphosphatase (ATPase) histochemistry or by immunoperoxidase labelling with antibodies directed at myosin heavy chain isoforms. ATPase histochemistry can be temperamental and lacks specificity, and both techniques require staining of serial cryosections to reveal the entire fibre type compliment of a single sample, which is time consuming and prone to inaccuracy. Here we describe an immunofluorescence labelling technique that enables rapid, accurate and specific identification of the 3 mature equine muscle fibre types in a single cryosection.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection

Tulloch

Perkins

Piercy³

2011

Equine Veterinary Journal

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“…Biopsy samples are best preserved when placed in an empty sterile pot and transported immediately on ice packs to the laboratory (Stanley et al . ). It is recommended to liaise with the diagnostic laboratory prior to collecting the biopsy and avoid posting samples at the end of the week to avoid unnecessary delays in transport.…”

Section: Diagnosismentioning

confidence: 97%

“…Skeletal muscle biopsy samples should be harvested from the gluteal or semi-membranosis muscles and are easily obtained from the standing sedated animal (Ledwith and McGowan 2004). Biopsy samples are best preserved when placed in an empty sterile pot and transported immediately on ice packs to the laboratory (Stanley et al 2009a). It is recommended to liaise with the diagnostic laboratory prior to collecting the biopsy and avoid posting samples at the end of the week to avoid unnecessary delays in transport.…”

Section: Diagnosismentioning

confidence: 99%

Polysaccharide storage myopathy – the story so far

Naylor

2015

Equine Veterinary Education

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Summary Polysaccharide storage myopathy (PSSM) was first described in 1992 in Quarter Horses, Appaloosa and Paint‐related breeds with clinical signs of exertional rhabdomyolysis. The disease is characterised by the accumulation of excessive glycogen and diastase‐resistant amylopectin polysaccharide inclusions within skeletal muscle fibres. The discovery of a mutation in the glycogen synthase 1 (GYS1) gene in some, but not all, horses with the disease suggested that PSSM represents a group of diseases with similar pathology but different aetiologies and that the pathogenesis is more complex than initially thought. Type 1 PSSM (PSSM1) refers to horses with the GYS1 mutation and has subsequently been identified in a large number of breeds found in Europe and North America. Clinical presentations associated with PSSM1 can vary and increased muscle enzyme activity at rest or following exercise often accompanies PSSM1; however, such changes may not be present in all cases. A diagnosis of PSSM is made on the basis of histopathology or specifically PSSM1 is diagnosed by genotyping horses for the GYS1 mutation. Cases usually respond well to management changes, in particular a diet low in starch and high in fat when it is accompanied by regular exercise.

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“…moistened gauze in a Petri dish at low temperatures [Dubowitz and Sewry, 2007], on damp gauze [Bergmann et al, 2009;Stanley et al, 2009] or even on dry gauze [Stanley et al, 2009]. In order to investigate the effects of new pharmacological approaches to restore impaired neuromuscular functions, e.g.…”

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confidence: 99%

Evaluation of Functional and Structural Alterations in Muscle Tissue after Short-Term Cold Storage in a New Tissue Preservation Solution

Wille

Gonder

Thiermann

et al. 2011

Cells Tissues Organs

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Storage of muscle preparations in vitro is required for the diagnosis of neuromuscular disorders and for electrophysiological tests. The current standard protocols for muscle storage or transport, i.e. placement on 0.9% NaCl-moistened gauze, lead to impaired function and structural alterations. For other tissues, however, improved preservation methods and solutions have recently been described. In this study, functional and structural alterations in the murine diaphragm were compared after storage on 0.9% NaCl-moistened gauze and after storage in different modifications of the new vascular preservation solution TiProtec®. Muscle force generation after nerve stimulation, histological parameters and ATP levels were investigated after 2.5 h of cold storage at 4°C in the different media and 0.5 h of rewarming at 25°C in Tyrode buffer. Murine diaphragms were injured during cold storage and rewarming, with the degree of the alteration being dependent on the type of solution used. There were no histological alterations and no caspase 3 activation in all groups. In contrast, diaphragms stored in the modified TiProtec solution showed markedly better performance concerning force generation after nerve stimulation (7.1 ± 1.1 cN · s) as well as higher ATP content (2.4 ± 0.7 µmol/g) and were superior to storage on 0.9% NaCl-moistened gauze (1.4 ± 0.4 cN · s; 0.3 ± 0.1 µmol/g). In conclusion, the modified TiProtec preservation solution showed promising results for short-term cold storage of murine diaphragms. For further evaluation, the transferability of these positive findings to storage conditions for muscles of other species, especially human muscle tissue, needs to be investigated.

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Storage‐associated artefact in equine muscle biopsy samples

Cited by 9 publications

References 14 publications

Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection

Multiple immunofluorescence labelling enables simultaneous identification of all mature fibre types in a single equine skeletal muscle cryosection

Polysaccharide storage myopathy – the story so far

Evaluation of Functional and Structural Alterations in Muscle Tissue after Short-Term Cold Storage in a New Tissue Preservation Solution

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