Effect of Freezing on Gene Expression of Gallus Sperm Based on Expression Profile Microarray
SU Yun-Ze1, XING Kai1,*, QI Xiao-Long1,*, GUO Yong1, NI He-Min1, SHENG Xi-Hui1, CHEN Yu2, WANG Liang2, WANG Xiang-Guo1, XIAO Long-Fei1
1 Animal Science and Technology College, Beijing University of Agriculture, Beijing 102206, China; 2 Beijing General Station of Animal Husbandry Service, Beijing 100107, China
Abstract：Cryopreservation of poultry sperm has great potential in artificial insemination, breeding selection and disease prevention. Although cryopreservation measures have been continuously optimized, the fertilization ability of spermatozoa after cryopreservation is still lower than that of fresh sperm. The purpose of this study was to investigate the difference of gene expression between fresh and thawed Gallus sperm. The gene expression levels in fresh and frozen sperm were detected by Affymetrix chicken genome-wide expression profile chip. The signal pathways and candidate genes related to freezing injury of chicken sperm were screened, and some differentially expressed genes (DEGs) were verified by qRT-PCR. The results showed that 687 DEGs were between the 2 groups (|Log2(FC)|>1; q-value<0.05). Among these genes, 615 were significantly upregulated and 72 were significantly downregulated in the fresh sperm group. Gene Ontology (GO) analysis showed that 196 DEGs of all significantly regulated genes were involved in mitogen-activated protein kinase (MAPK) signal pathway, ATP activation, cell cycle and other biological processes. KEGG analysis showed that DEGs were significantly (P<0.01) enriched on carbohydrate metabolism, signal transduction, cell growth and death. Four DEGs (cold-inducible RNA-binding protein (CIRBP), 90 kD heat shock protein (HSP90), HSP70 and Ras homolog family member A (RHOA)) were selected and verified by qRT-PCR,and the results were in accordance with the above expression profile. Therefore, MAPK pathway, cell cycle regulation pathway and CIRBP, HSP90, HSP70 and RHOA genes may be important regulatory pathways and candidate genes leading to significant differences in cryopreservation. This study would provide a theoretical reference for understanding the potential molecular mechanism of frozen-thawed chicken sperm damage and how to improve the fertility of frozen-thawed chicken sperm.
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