title: Assessing the reproducibility of fractional rates of protein synthesis in muscle tissue measured using the flooding dose technique.
abstract: The flooding dose technique of Garlick et al. (1980) has become the main method for measuring tissue and whole-animal rates of protein synthesis in ectotherms. However, single tissue samples are used to determine rates of protein synthesis and no studies have examined the pattern of flooding in large tissues such as the white muscle in fishes, which can comprise up to 55% of the wet body mass of a fish and which is poorly perfused. The present study has examined, for the first time, the patterns of flooding and measured rates of protein synthesis in five different regions of the white muscle in the Arctic charr Salvelinus alpinus ranging in size from 25g to 1.6kg following a flooding dose injection of L-[(3)H]-phenylalanine. The results indicate that the degree of flooding (i.e. free pool specific radioactivity relative to that of the injection solution) and elevation in free phenylalanine concentrations can vary between regions but the calculated fractional rates of protein synthesis were similar in four of the five regions studied. The variability in rates of protein synthesis increased with body size with greater variability observed between regions for fish >1kg in body mass. For consistency between studies, it is recommended that samples are taken from the epaxial muscle in the region below the dorsal fin when measuring fractional rates of white muscle synthesis in fishes.
authors.affiliation: 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.
authors.forename: Ian D
authors.initials: ID
authors.lastname: McCarthy
authors.affiliation: School of Biological Sciences, College of Natural Sciences, Bangor University, Deiniol Road, Bangor, Gwynedd, LL57 2UW, UK.
authors.forename: James
authors.initials: J
authors.lastname: Brown
mesh_terms: D000818:Animals; D007700:Kinetics; D009124:Muscle Proteins / Q000096:biosynthesis*; D010649:Phenylalanine / Q000008:administration & dosage*; D015203:Reproducibility of Results; D014337:Trout
chemical_list: D009124:Muscle Proteins; D010649:Phenylalanine
keywords: Arctic charr; Flooding dose technique; Protein synthesis; Repeat measures; Reproducibility; Salvelinus alpinus; White muscle