Proteome analysis of Oncorhynchus species during embryogenesis

Abstract

To understand the molecular mechanisms underlying normal and abnormal development of two salmonids, masu salmon (Oncorhynchus masou) and rainbow trout (O. mykiss), we used two-dimensional (2-D) electrophoresis to construct a series of 2-D maps during the embryonic period. We identified all visible protein spots on the 2-D map by assigning numbers for masu salmon and rainbow trout, and we determined N-terminal sequences of proteins for one hundred of the spots, that appear at very high concentrations in the whole embryos of masu salmon and rainbow trout. We also characterized embryonic stages according to the periods of appearance of spots. Most of the N-terminal sequences were identical or at least highly similar to partial sequences reported for vitellogenin (Vtg) of O. mykiss. A potential proteolytic processing of Vtg for rainbow trout is discussed in relation to the time of appearance and relative position of Vtg fragments within the complete protein sequence.

Prediction of Proteolytic Process Based on N-Terminal Sequences and Molecular Weights by Proteomics and Proteome Analysis

“…A series of 2-D maps constructed for respective embryos of O. masou during the period from fertilization to hatching were previously reported 9 . To date, we determined N-terminal sequences for 48 spots which are identical to, or highly similar to, partial sequence of Vtg of O. mykiss 20 (Table 1).…”

“…Proteins were directly excised from a PVDF membrane, and N-terminal sequences were determined. Detail experimental conditions of 2-DE and N-terminal sequencing are described in 8,9 .…”

“…Using 2-DE to construct a series of 2-D maps in O. masou during the embryogenic period, we detected synchronous increments in protein constituents during the following periods of embryogenesis: (i) the morphological development corresponding to the shift from principal events of axis formation to differentiation of organs and tissues, (ii) retinal pigmentation, and (iii) immediately before hatching 8 . Proteolytic process in cells is emerging as an important mechanism for controlling cellular processes such as cell-cycle regulation by degrading cyclins, stress response by 20S proteasomes 10 , ubiquitin-dependent protein degradation 11 , apoptosis by proteolytic cleavage of certain cellular proteins by caspases 12 , and degradation of yolk proteins during embryonic development in O. mykiss 9,13,14 , Xenopus laevis 15 , the silkmoth Bombyx mori 16 , orthopteran Blattella germanica 17 , stick insect Carausius morosus 18 and cricket Acheta domesticus 19 . In proteomics and proteome analysis using 2-DE, molecular weights and N-terminal sequences for individual protein spots are relatively easily determined.…”

Egg protein bound amino acid content and embryo/larva success in common dentex (Dentex dentex), a marine pelagophil teleost

“…(6) Because the vast majority of total proteins in fish post-vitellogenic oocytes and ovulated eggs are Vtg-derived proteins (e.g., see Kanaya et al, 2000;Tay et al, 2006), their dominant MS/MS spectra make it difficult to identify other, rare proteins. It is therefore highly probable that the PAAs measured in the current study are Vtg-derived PAAs.…”

Proteomic analysis of inviable salmonid hybrids between female masu salmon Oncorhynchus masou masou and male rainbow trout Oncorhynchus mykiss during early embryogenesis

“…In zebrafish Danio rerio (Hamilton 1822), profiles of protein expression were examined during early development using 2‐DE and MS analysis (Tay et al , 2006). In salmonids, proteome analysis in the embryogenesis of O. m. masou and O. mykiss also was performed using 2‐DE and N‐terminal sequencing (Kanaya et al , 2000).…”