Proteome Comparative Analysis Of Gynogenetic Haploid And Diploid Embryos Of Goldfish (Carassius Auratus)

It was found recently that in the gynogenetic haploid and diploid embryos of goldfish, which have exactly the same genome, the haploid condition results in obstruction of gene expression and abnormal development while the diploid embryos have normal gene expression and development. A diploid-dependent regulatory apparatus was proposed by Prof. Luo et al to regulate gene expression. The haploid condition results in obstruction of gene expression and abnormal development because the diploid-dependent regulatory apparatus will regulate gene expression in the haploid embryos according to the same rule as in the diploid embryos. To study the difference at the protein expression level of the embryos of haploid and diploid in development and to identify the important proteins associated with the embryonic development, we extracted the total proteins of both the gynogenetic haploid and diploid embryos of goldfish in the same eye formation stages (divided into the substages of HE-1 and DE-1, HE-2 and DE-2, HE-3 and DE~3 respectively). Two-dimensional polyacrylainide gel electrophoresis was used to separate proteins. The stained gel images were analyzed with the PDQUEST software. Most proteins are distributed in pH 5-10and MrlOOOO-SOOOODa. We found more than 100 different proteins in HE-3 and DE-3, and more than 250 different proteins in HE-2 and DE-2. 70 protein spots that were differentially expressed in haploid and diploid embryos respectively were identified by matrix-assisted laser desorption/ionizalion time~of-flight mass spectrometry and database analysis.In HE-3 and DE-3, 16 protein spots that were absolutely different (only expressed in diploid embryos but not in haploid embryos or vice versa) and 16 protein spots that were up- and down-r gulated were identified unambiguously. In HE-2 and DE-2, 20protein spots that were absolutely different (only expressed in diploid embryos but not in haploid embryos or vice versa) and 5 protein spots that were up- and down-regulated were identified unambiguously, some of these different proteins are correlative with eyes development, nerve development, development regulation, cell differentiation and maintenance, arthromere formation and signal transduction. It shown that gene expression have significant difference between diploid and haploid during embryos developing. These differences may cause the abnormal development of haploid embryos.We also analyze comparatively the proteome of the second andthird stage, including DE-3 and DE-2; HE~3 and HE-2. Significant differences were found in the second and third stage, and some differently expressed proteins were analyzed.