Pkg2在小鼠中的年龄依赖性表达特征分析

Study on age-dependent expression of Pgk2 in mice

目的利用基因芯片技术筛选与精子发生相关的基因,并研究其表达特点。方法将4d、9d、18d、35d、54d龄和6月龄小鼠睾丸组织cDNA探针与Affymetrix全基因组芯片进行杂交,筛选出差异表达的基因。然后通过RT-PCR分析差异表达基因在小鼠睾丸不同发育阶段中的表达。结果对Affymetrix全基因组芯片杂交结果分析后,筛选得到1个差异表达杂交点,通过在NCBI网站与小鼠全基因组序列Blast分析可知该差异表达基因是Pgk2基因。该基因在4d、9d、1 8d、35d、54d龄和6月龄小鼠睾丸中信号强度分别是1.7(A)、161.5(P)、634(P)、1881.3(P)、3123.7(P)和3440.4(P)表明Pgk2基因在4d龄小鼠睾丸中无表达,9d低表达,从18年龄后开始高表达;RT-PCR结果表明:小鼠Pgk2基因在小鼠9d龄及之前的睾丸中无表达或低表达,在18d龄睾丸后开始高表达,与芯片数据的结果相一致。结论 Pgk2基因为小鼠年龄依赖性表达基因,推测其在精子发生过程中有重要作用。

Objective To screen spermatogenesis associated genes using cDNA microarray and analyze their expression pattern. Methods Testis tissues of Balb/C mice with different postnatal days Were collected for spermatogenesis associated genes screening using mouse whole genome Affymetrix chip. The characteristics of the selected genes were analyzed by various bioinformatics tools. The expression of differential spermatogenesis associated genes in mice testis were further confirmed by RT-PCR. Results A highexpression gene Pgk2 was detected in advanced stage of mouse testis. The hybridization signal intensities of gene Pgk2 in testis tissues on postnatal d 4, d 9, d 18, d 35, d54, and 6 months were 1.7 （Absent expression, A）, 161.5 （Present expression, P）, 634 （P）, 1881.3（P）, 3123.7 （P） and 3440.4（P）, respectively. The expression of gene Pgk2 was not detected on d4 and then it expression was gradually increases from on d9 to 6 months. Similar expression pattern of gene Pgk2 was further confirmed by RT-PCR. Conclusion Pgk2 might be a age-dependent expression gene of mouse. It might be associated with male spermatogenesis.