Immunofluorescent visualisation of focal adhesion kinase in human skeletal muscle and its associated microvasculature

Wilson, Oliver J., Shaw, Christopher S., Sherlock, Mark, Stewart, Paul M. and Wagenmakers, Anton J. M. 2012, Immunofluorescent visualisation of focal adhesion kinase in human skeletal muscle and its associated microvasculature, Histochemistry and cell biology, vol. 138, no. 4, pp. 617-626, doi: 10.1007/s00418-012-0980-x.

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Title Immunofluorescent visualisation of focal adhesion kinase in human skeletal muscle and its associated microvasculature
Author(s) Wilson, Oliver J.
Shaw, Christopher S.ORCID iD for Shaw, Christopher S.
Sherlock, Mark
Stewart, Paul M.
Wagenmakers, Anton J. M.
Journal name Histochemistry and cell biology
Volume number 138
Issue number 4
Start page 617
End page 626
Total pages 10
Publisher Springer
Place of publication Heidelberg, Germany
Publication date 2012-10
ISSN 0948-6143
Keyword(s) focal adhesion kinase
skeletal muscle
confocal imaging
Focal Adhesion Protein-Tyrosine Kinases
Microscopy, Fluorescence
Muscle Fibers, Skeletal
Muscle, Skeletal
Young Adult
Summary Within animal skeletal muscle, focal adhesion kinase (FAK) has been associated with load-dependent molecular and metabolic adaptation including the regulation of insulin sensitivity. This study aimed to generate the first visual images of the localisation of FAK within human skeletal muscle fibres and its associated microvasculature using widefield and confocal immunofluorescence microscopy. Percutaneous muscle biopsies, taken from five lean, active males, were frozen and 5-μm cryosections were incubated with FAK antibodies for visualisation in muscle fibres and the microvasculature. Anti-myosin heavy chain type I was used for fibre-type differentiation. Muscle sections were also incubated with anti-dihydropyridine receptor (DHPR) to investigate co-localisation of FAK with the t-tubules. FITC-conjugated Ulex europaeus Agglutinin I stained the endothelium of the capillaries, whilst anti-smooth muscle actin stained the vascular smooth muscle of arterioles. Fibre-type differences in the intensity of FAK immunofluorescence were determined with image analysis software. In transversely and longitudinally orientated fibres, FAK was localised at the sarcolemmal regions. In longitudinally orientated fibres, FAK staining also showed uniform striations across the fibre and co-staining with DHPR suggests FAK associates with the t-tubules. There was no fibre-type difference in sarcoplasmic FAK content. Within the capillary endothelium and arteriolar smooth muscle, FAK was distributed heterogeneously as clusters. This is the first study to visualise FAK in human skeletal muscle microvasculature and within the (sub)sarcolemmal and t-tubule regions using immunofluorescence microscopy. This technique will be an important tool for investigating the role of FAK in the intracellular signalling of human skeletal muscle and the endothelium of its associated microvasculature.
Language eng
DOI 10.1007/s00418-012-0980-x
Field of Research 111699 Medical Physiology not elsewhere classified
1199 Other Medical And Health Sciences
Socio Economic Objective 970111 Expanding Knowledge in the Medical and Health Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
ERA Research output type C Journal article
Copyright notice ©2012, Springer
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Document type: Journal Article
Collections: Faculty of Health
School of Exercise and Nutrition Sciences
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