Proteomic analysis of muscle between hybrid abalone and parental lines Haliotis gigantea Reeve and Haliotis discus hannai Ino

Abstract

To understand the potential molecular mechanism of heterosis, protein expression patterns were compared from hybrids of Haliotis gigantea (G) and Haliotis discus hannai (D) using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight analyses. Expression differences were observed in muscle samples from the four groups with 673 ± 21.0 stained spots for H. discus hannai ♀ × H. discus hannai ♂ (DD), 692 ± 25.6 for H. gigantea ♀ × H. gigantea ♂ (GG), 679 ± 16.2 for H. discus hannai ♀ × H. gigantea ♂ (DG) (F 1 hybrid) and 700 ± 19 for H. gigantea ♀ × H. discus hannai ♂ (GD) (F 1 hybrid). Different 2-DE image muscle protein spots had a mirrored relationship between purebreds and the F 1 hybrid, suggesting that all stained spots in F 1 hybrid muscle were on 2-DEs from parents. DD and DG clustered together first, and then clustered with GD, whereas the distance of DD and GG was maximal according to hierarchical cluster analysis. We identified 136 differentially expressed protein spots involved in major biological processes, including energy metabolism and stress response. Most energy metabolism proteins were additive, and stress-induced proteins displayed additivity or overdominance. In these 136 identified protein spots, hybrid offspring with additivity or over-dominance accounted for 68.38%. Data show that a proteomic approach can provide functional prediction of abalone interspecific hybridization.

Evaluation of the Use of TRIzol-Based Protein Extraction Approach for Gel-Based Proteomic Analysis of Dried Seafood Products and Chinese Tonic Foods

“…Given that each protein extraction method has its own bias in extracting proteins; it is not surprising that a substantial amount of differing protein spots were found between the gels from the two methods. A study using TRIzol extraction demonstrated approximately 700 protein spots on the 2-DE gel of fresh abalone muscle [ 35 ]. However, only approximately 200 protein spots were obtained from the dried abalone sample in the current study ( Figure 2 ).…”

“…The TRIzol-based extraction method was first applied successfully on halophilic protein samples for the generation of 2-DE in 2006 [ 19 ]. This protein extraction method has subsequently been widely used in 2-DE for a wide range of samples, including rat spinal cord [ 20 ], human/animal cell lines [ 21 , 22 , 23 ], dinoflagellates [ 11 , 24 , 25 , 26 , 27 , 28 ], mites [ 29 ], plant tissue [ 30 ], clinical samples [ 31 , 32 , 33 ], marine animals [ 34 , 35 , 36 ], and reef corals [ 37 ] ( Table 1 ). This TRIzol-based method has gained popularity for 2-DE because it offers several advantages [ 19 , 24 ].…”

Proteomic profiling of eggs from a hybrid abalone and its parental lines: Haliotis discus hannai Ino and Haliotis gigantea

“…Protein extractions were performed using TRIzol-mediated sample preparation as described previously (Di et al 2013(Di et al , 2015. The protein pellet was air-dried and resuspended in isoelectric focusing (IEF) redissolving buffer [7 M of urea, 2 M of thiourea, 4% (w/v) CHAPS, 40 mM of Tris].…”

Protein changes in abalone foot muscle from three geographical populations of Haliotis diversicolor based on proteomic approach

“…Protein identification was carried out as previously described (Di et al 2013(Di et al , 2015. Briefly, spots of interest were excised and trypsin digested trypsin, with 0.5 lL of extracted sample analyzed by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) with a 5800 Proteomics Analyzer (Applied Biosystems, CA, USA).…”