The Influence Of AC3 Deletion On Apoptosis In The Testis Of Mouse And The Analysis Of Differentially Expressed Genes

The second messenger of cyclic adenosine-3'-5' –mono-phosphate(cAMP) is a kind of general broad-spectrum intracellular messenger, it plays an extensive role in various biological systems. Adenylate cyclase(AC3) is the main membersfor cAMP signaling pathways, its role is to ATP into cAMP. AC3 deletions damaged the cAMP signal pathway, there were anosmia, along with the ability of attack male, maternity care, learning, memory and reproductive occurred abnormalities. Spermatogenesis is a highly precise production process of mature sperm undergo mitosis, meiosis, and spermiogenesis(spermiogenesis) three stages. Transcription factor regulation of the specific stage of gene expression plays an important role in regulating in this process. Some of the most important changes occurred in the sperm formation in the early stage, the mechanism of transcription is also the most active at this time. AC3 at this stage to express, through the AC3 inactivation of genes determine its importance in the process of spermatogenesis is very necessary.The experimental material are AC3 wild and knockout mice testis, at the first we use TUNEL staining to detect apoptosis of testicular tissue in mice, the results show that the AC3 missing cell apoptosis in the testicular tissue of mice increased significantly. second, gene microarrays were used to screen differentially expressed genes in testis from the AC3 knockout(AC3-/-) and the wild-type(AC3+/+) mice. The differentially expressed genes identified were subjected tobioinformatic analysis and verified by qRT-PCR. Results show that the gene of 693 had greater than 2-fold differential expression between AC3-/- and AC3+/+ mice, Of these genes, 330 were up-regulated, and 363 were down-regulated. To analysis signaling pathways and functional annotation of the differentially expressed genes, GO annotation analysis showed that 242 differentially expressed genes related to molecular function, 461 differentially expressed genes involved in biological processes, 516 differentially expressed genes involved in cell component function. Gene Ontology(GO) analysis showed that these differentially expressed genes were involved transcription activity, defense reaction, cell adhesion, cell death, immune response. Pathway analysis results show that the AC3 knockout, 34 of the signal path has changed, including calcium signaling Pathway that seven gene are significant changes, the olfactoryreceptors signaling pathways of significant changes include 28 genes, axons guidance signaling pathways in significant changes include 6 gene and 3 cell junction related signaling Pathway, a total of 25 genes changed. In this paper, quantitative RT-PCR verification of the differentially expressed epigenetic related genes. Including: cAMP signal pathway members calml3, olfr1016, olfr1273, olfr711; Development and cell death related gene crisp1, ywhaz, irs4, olig2 prevented, cyplla1; Participate in the epigenetic regulation of related gene mettl14, dntt; Participate in sperm movement and capacitation related gene pxn, defb22, snca, dcc, itgav. qRT-PCR results showed which was consistent with results of the microarray screening and were significantly changed. In addition, the chip results also found a lot of the expression of genes related to cell death also significant changes. The results has the important enlightenment for the understanding of mechanism of male sterility in AC3-/- mice.