Eastern grey kangaroo (Macropus giganteus) myofibres. 1. A simplified classification method using two commercially available antibodies
N. B. Spiegel A B E , W. H. Johns C , S. D. Sinclair C , P. C. Wynn C D , J. M. Thompson A and P. L. Greenwood C FA School of Environmental and Rural Science, University of New England, Armidale, NSW 2351, Australia.
B Faculty of Veterinary Science, University of Sydney, Sydney, NSW 2006, Australia.
C Industry & Investment NSW, University of New England, Armidale, NSW 2351, Australia.
D E. H. Graham Centre for Agricultural Innovation, Charles Sturt University, Wagga Wagga, NSW 2678, Australia.
E Present address: School of Environmental Science, Murdoch University, Murdoch, WA 6150, Australia.
F Corresponding author. Email: paul.greenwood@industry.nsw.gov.au
Animal Production Science 50(6) 386-392 https://doi.org/10.1071/AN09195
Submitted: 11 December 2009 Accepted: 14 April 2010 Published: 11 June 2010
Abstract
Skeletal muscles from eastern grey kangaroos (Macropus giganteus) were assessed for myofibre contractile and metabolic characteristics using immunocytochemical and histological staining of serial sections. Myofibre classification using monoclonal antibodies that typically bind to mammalian slow (clone WB-MHC), fast (clone MY-32) and Types 1, 2X and 2B (clone S5 8H2) myosin heavy chains was validated using acid- and alkali-preincubated myofibrillar ATPase, NADH and α-glycerophosphate dehydrogenase stains. Myofibres were classified as Type 1 (slow oxidative), Type 2A (fast oxidative-glycolytic), Type 2X/2B (fast glycolytic) or intermediate or transitional myofibre Types 2C (Type 1–Type 2A intermediate) and 2AX/B (Type 2A–Type 2X/2B intermediate). The Type 2 (fast) antibody (clone MY-32) used in the present study did not bind to a subset of fast myofibres in any of the eight kangaroo muscles investigated. These myofibres were identified as Type 2A using clone S5 8H2 and on the basis of the histochemical staining profile. Hence, a simplified immunostaining system using only clones WB-MHC (anti-Type 1) and MY-32 (anti-Type 2X/2B) allowed five myofibre types to be identified based on the staining pattern and intensity of staining for the two antibodies. It is concluded that the myofibres of muscles from kangaroos can be quickly classified into five types using two commercially available antibodies. This method is directly applicable for routine investigations into the myofibre properties of commercially important muscles from the kangaroo musculature and, when combined with enzymatic assays for oxidative and glycolytic activity, will allow for a better understanding of factors influencing the quality of meat from kangaroos.
Additional keyword: macropod.
Acknowledgements
Financial support was provided by the Rural Industries Research and Development Corporation (RIRDC) and the Kangaroo Industry Association of Australia (KIAA). The assistance provided by technical staff of the NSW Department of Primary Industries Beef Industry Centre of Excellence and Dr Ian Colditz (CSIRO, Armidale, NSW, Australia) is also gratefully acknowledged. Constructive editorial comments provided by Dr Paul Hopwood during the preparation of this manuscript are greatly appreciated. The authors also acknowledge Dr Brigitte Picard (Institut National de la Recherch Agronomique) who provided antibody S5 8H2.
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