| In our experiment, myometirum and ovary are collected from nonpregnant (NP, n=3), early pregnant (EP, 25d, n=3), late pregnant (LP, 85d, n=3) large-white pigs which the NP pigs were used as contract. Then the RNA was extracted and hybridized with Affymetrix Genechip Porcine Genome. After normalization, differential expressed genes were filtered under 2-fold standard. Cluster, GO and pathway analysis were done. And, at last the gene regulatory network was constructed, and new candidate reproductive genes were selected. The results are as below:1) There are 974 (418 up and 556 down) and 1800(826 up and 974 down) 2-fold differential display genes in myometrium respectively in EP and LP periods, 335 (189 up and 146 down) and 710 (236 up and 487 down) 2-foled differential display genes in ovary respectively in EP and LP periods.2) And as we consider the NP periods as quiescence status, the EP & LP periods as startup status, 7 clusters can be classified use SOM method. Using this cluster results we chose 33 genes for forward selection of new candidate gestation genes.3) According to the gene ontology analysis, in the field of molecular function,the genes with MHC class II receptor activity and enzyme regulator activity in LP period are not differentially expressed, the genes with MHC class I receptor activity in EP period are not differentially expressed, and however, iron ion binding genes only differentially expressed in myometirum. Compared with EP period, the expression of the genes in ovary with lipid binding, endopeptidase activity, chemokine activity have changed distinctly; in the field of Biological Process, in LP period ovary, the expression of genes with normal biological process changed little, and the genes associate with stimulus & stress response changed more than EP period ovary and EP& LP period myometirum. In EP period myometirum, genes related with cell growth, regulation of cell size, cell cycle have differential expression; in the field of Cellular Component, the gene have differential expression are all related with molecular membrane, in LP period ovary and EP & LP myometirum, the genes associated with plasma membrane, MHC protein complex, immunological synapse, protein complex have changed expression. And the genes which related with membrane integral and intrinsic have largest changed expression.4) By the analysis of the genes which are differentially expressed in ovary, we find no valid pathways; but in the analysis of the genes which are differentially expressed in myometirum, we found 5 valid pathways in early pregnant period and 3 valid pathways in late pregnant period.5) After networks learing with simulative data, we found out a suitable network for our data, and a network with many small networks was reconstructed.6) Using the networks we constructed and the information we can mined from the bioinformatics analysis, the significant function of ESR, IGFBP6, UF, IGF2, RBP4 are confirmed, and also we selected several new candidate reproductive genes which are Ssc.204, Ssc.315, Ssc.275, Ssc.256, et al., and we have inferred their function. |
