Proteomic investigation of the effects of weight loss in the gastrocnemius muscle of wild and NZW rabbits via 2D‐electrophoresis and MALDI‐TOF MS

Abstract

The study of changes within the key agents regulating metabolism during genetic upgrading because of selection can contribute to an improved understanding of genomic and physiological relationships. This may lead to increased efficiencies in animal production. These changes, regarding energy and protein metabolic saving mechanisms, can be highlighted during food restriction periods. In this study, a 20% weight reduction was induced in two rabbit breeds: New Zealand white, a selected meat producer (Oryctolagus cuniculus cuniculus), and Iberian wild rabbit (Oryctolagus cuniculus algirus), with the aim of determining differential protein expression in the gastrocnemius muscle within control (ad libitum) and restricted diet experimental animal groups, using techniques of two-dimensional gel electrophoresis and peptide mass fingerprinting. Results show that L-lactate dehydrogenase, adenylate kinase, beta enolase and alpha enolase, fructose bisphosphate aldolase A and glyceraldehyde 3-phosphate dehydrogenase, which are enzymes involved in energy metabolism, are differentially expressed in restricted diet experimental animal groups. These enzymes are available to be further tested as relevant biomarkers of weight loss and putative objects of manipulation as a selection tool towards increasing tolerance to weight loss. Similar reasoning could be applied to 2D gel electrophoresis spots corresponding to the important structural proteins tropomyosin beta chain and troponin I. Finally, a spot identified as mitochondrial import stimulation factor seems of special interest as a marker of undernutrition, and it may be the object of further studies aiming to better understand its physiological role.

Pig proteomics: A review of a species in the crossroad between biomedical and food sciences

“…On the other hand, the individuation of pyruvate kinaseM1 and tropomyosin levels in LW was related to water holding capacity and meat color at 24 h after slaughter. A bioinformatic integrated analysis strengthened the correlation between over-expression of structural proteins in Large White and higher growth aptitude in this breed, a result that may be found in other species such as the rabbit [44] or cattle [45]. On the other hand, enzymes taking part into branching glycolytic reactions, such as glycerol 3-phosphate and creatine kinase M, were related to higher lipogenesis and slower albeit prolonged glycolytic rate in the Casertana breed [42].…”

Mass spectrometry and animal science: Protein identification strategies and particularities of farm animal species

“…Recent interesting examples include the study of Longissimus thoracis post-mortem proteome evolution in cattle [115] or the search in swine for indicators of an appropriate water holding capacity thus avoiding PSE -Pale Soft and Exudative and DFD -Dark Firm and Dry meats [116]. Interestingly, in the bovine case [101], authors were able to successfully identify 26 proteins out of 35 candidates, an approximate success rate of 75%, slightly lower than a PMF-based identification strategy such as the one mentioned for the rabbit study [103] that attained a degree of success of approximately 85%. These results seem therefore to point out that PMF can be used as a valid identification strategy for well-known samples of highly conserved proteins.…”

“…Although less common in other areas of proteomics, it is still very popular in farm animal muscle research, as witnessed by the number of recent publications that resort to PMF [35,36,103]. In these cases, relatively high success rates were obtained with the PMF identification strategy, as most of the proteins selected for identification were successfully identified.…”

“…In these cases, relatively high success rates were obtained with the PMF identification strategy, as most of the proteins selected for identification were successfully identified. The degree of success of this approach is likely to be directly related to the fact that the species studied: rabbit [103], pig [35] or chicken [36] are either well described or sequenced. Nevertheless, and in the case of rabbit [103] a non-sequenced organism, several proteins were identified in other organisms with special reference to laboratory rats (Rattus norvegicus) and mice (Mus musculus).…”

“…The combination of 2DE and visible staining systems has been thoroughly used in farm animal muscle proteomics. Examples include the study of bovine and ovine muscle hypertrophy characterization [101,102] or the comparison of two rabbit breeds when subjected to undernutrition [103] and studies involving other species, as cattle [104,105], swine [106] or poultry [107]. Due to the low sensitivity and lack of reproducibility of these systems they have been replaced over the last years with differential proteomic assays based on fluorescent dyes, namely DIGE [32].…”