Proteome Comparative Analysis Of The Developmental Process Of Gynogenetic Goldfish Embryos And Optimization Of The Method For Peptide Mass Fingerpriting Analysis

Part one: Previous results indicated that the abnormal development of haploid goldfish embryos is due to the expression obstruction of some proteins, which are correlative with eyes development, nerve development, development regulation, cell differentiation and maintenance, arthromere formation and signal transduction.To better understand the molecular events underlying the processes occurring at the embryos developing of gynogenetic haploid and diploid and elucidate the mechanism of abnormal development in haploid embryos, we conducted a proteome comparative analysis of the gynogenetic haploid and diploid embryos of goldfish in the same tail fin appearance stages (31hrs after fertilization). Protein profiles of embryos among the 3 stages (HE-1 and DE-1, HE-2 and DE-2, HE-3 and DE-3 represent 31hrs, 33s, 35hrs after fertilization respectively) exhibited a number of stage-specific changes. Identification of developmentally related proteins by MALDI-TOF and ESI-Q-TOF revealed several groups of functionally related proteins over the period HE1 —3, including three proteins associated with eyes development (Gamma-crystallin Ml-1, Gamma crystalline M3, homeodomain protein (clone G6)) , seven development regulated proteins(for example, insulin-like growth factor binding protein-2, Smad2, Methy1-CpG binding protein 2), four proteins correlated with nerve development (for example, nerve growth factor beta chain precursor , Neural plakophilin-related arm-repeat protein). Furthermore, some proteins associated with cell differentiation and maintenance, arthromere formation and signal transduction were also identified.Luo et al have proposed that the cause of abnormal development in haploid embryos is that there is a diploid-dependent regulatory apparatus which modulates gene expression. We found that the down-regulation of some proteins is likely because of the turbulence of gene expression in haploid embryos developmental process, resulting in the abnormal development of organ and morphology. In addition, the up-regulation of some proteins indicates the formation of some tissue and organ in the goldfish tail fin appearance stages.