Validation of the flooding dose technique to determine fractional rates of protein synthesis in a model bivalve species, the blue mussel (Mytilus edulis L.).
For the first time, use of the flooding dose technique using (3)H-Phenylalanine is validated for measuring whole-animal and tissue-specific rates of protein synthesis in the blue mussel Mytilus edulis (61mm shell length; 4.0g fresh body mass). Following injection, the phenylalanine-specific radioactivities in the gill, mantle and whole-animal free pools were elevated within one hour and remained elevated and stable for up to 6h following injection of (3)H-phenylalanine into the posterior adductor muscle. Incorporation of (3)H-phenylalanine into body protein was linear over time following injection and the non-significant intercepts for the regressions suggested incorporation into body protein occurred rapidly after injection. These results validate the technique for measuring rates of protein synthesis in mussels. There were no differences in the calculated rates following 1-6h incubation in gill, mantle or whole-animal and fractional rates of protein synthesis from the combined time course data were 9.5±0.8%d(-1) for the gill, 2.5±0.3%d(-1) for the mantle and 2.6±0.3%d(-1) for the whole-animal, respectively (mean values±SEM). The whole-animal absolute rate of protein synthesis was calculated as 18.9±0.6mg protein day(-1). The use of this technique in measuring one of the major components of maintenance metabolism and growth will provide a valuable and convenient tool in furthering our understanding of the protein metabolism and energetics of this keystone marine invertebrate and its ability to adjust and respond to fluctuations, such as that expected as a result of climate change.
School of Ocean Sciences, College of Natural Sciences, Bangor University, Askew Street, Menai Bridge, Anglesey, LL59 5AB, UK; Laboratorio de Manejo, Ecologia e Conservação Marinha, Instituto Oceanográfico, Universidade de São Paulo, Praça do Oceanográfico 191, Cidade Universitária, 05508-120 São Paulo SP, Brazil. Electronic address: i.mccarthy@bangor.ac.uk.
Ian D
ID
McCarthy
School of Ocean Sciences, College of Natural Sciences, Bangor University, Askew Street, Menai Bridge, Anglesey, LL59 5AB, UK.
Ruth
R
Nicholls
School of Ocean Sciences, College of Natural Sciences, Bangor University, Askew Street, Menai Bridge, Anglesey, LL59 5AB, UK.
Shelagh K
SK
Malham
School of Biological Sciences, College of Natural Sciences, Bangor University, Deiniol Road, Bangor, Gwynedd, LL57 2UW, UK.
Nia M
NM
Whiteley
D000818:Animals; D001835:Body Weight; D055868:Floods; D005880:Gills / Q000378:metabolism; D023421:Models, Animal; D049880:Mytilus edulis / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D014176:Protein Biosynthesis; D015203:Reproducibility of Results; D013045:Species Specificity; D013997:Time Factors
D010649:Phenylalanine
Bivalve; Flooding dose technique; Mussel; Mytilus edulis; Protein synthesis