CAJ | 학술논문

背景:精子发生过程受许多特异分子及细胞间作用的严格调节,包括染色体结构变化及一系列特定基因程序性表达调控.长期以来由于缺乏合适的体内和体外研究模型,精子发生分子机制的研究进展较慢,尤其缺乏对关键调节基因的认识.目的:筛选与精子发生相关的基因,并分析其表达特点.方法:将4,9,18,35,54 d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.然后通过反转录-聚合酶链反应分析差异表达基因在小鼠睾丸不同发育阶段中的表达.结果与结论:对Affymetrix全基冈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与小鼠全基因组序列Blast分析可知该差异表达基因是Tpap基因.该基因在4,9,18,35,54d和6月龄小鼠睾丸中杂交信号校正值分别是4.4(A)、12.9(A),262.4(P)、1 136.7(P)、1 617.5(P)和1 128(P),表明4,9 d小鼠睾丸中该基因无表达,18 d小鼠开始表达,RT-PCR结果表明小鼠Tpap基因在小鼠9 d及之前的睾丸中没有表达,在18 d睾丸后开始表达,与基因芯片分析相一致.结果表明,Tp印基因为小鼠年龄依赖性表达基因,小鼠Tpap的表达与小鼠精子发生的过程有很强的一致性,推测该基因在哺乳动物精子发生中起重要作用.
배경:정자발생과정수허다특이분자급세포간작용적엄격조절,포괄염색체결구변화급일계렬특정기인정서성표체조공.장기이래유우결핍합괄적체내화체외연구모형,정자발생분자궤제적연구진전교만,우기결핍대관건조절기인적인식.목적:사선여정자발생상관적기인,병분석기표체특점.방법:장4,9,18,35,54 d화6월령소서고환조직cDNA탐침여Affymetrix전기인조심편진행잡교,사선출차이표체적기인.연후통과반전록-취합매련반응분석차이표체기인재소서고환불동발육계단중적표체.결과여결론:대Affymetrix전기강조심편잡교결과분석후,사선득도1개차이표체잡교점.통과재NCBI여소서전기인조서렬Blast분석가지해차이표체기인시Tpap기인.해기인재4,9,18,35,54d화6월령소서고환중잡교신호교정치분별시4.4(A)、12.9(A),262.4(P)、1 136.7(P)、1 617.5(P)화1 128(P),표명4,9 d소서고환중해기인무표체,18 d소서개시표체,RT-PCR결과표명소서Tpap기인재소서9 d급지전적고환중몰유표체,재18 d고환후개시표체,여기인심편분석상일치.결과표명,Tp인기인위소서년령의뢰성표체기인,소서Tpap적표체여소서정자발생적과정유흔강적일치성,추측해기인재포유동물정자발생중기중요작용.
BACKGROUND: These serial processes for forming male gametes are basically controlled by the programmed expression of a number of stage-specific genes. However, many aspects of the mechanisms of spermatogenesis have remained elusive because of a lack of suitable in vitro or in vivo models.OBJECTIVE: To screen genes involved in spermatogenesis, and to analyze its expression characteristics. METHODS: Testes cDNA samples from Balb/C mice of different postnatal days (4,9,18,35, 54 days and 6 months, respectively) were hybridized with mouse whole genome Affymetrix chip to screen the testis-ralated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. RT-PCR was used here to identify the expression of the selected genes in mice testis.RESULTS AND CONCLUSION: The Affymetrix chip probe of mouse Tpap was graduated higher expression with developmental stages of mouse testis. The scaling hybridization signal intensities of the tested testis on days 4, 9,18, 35, 54, and 6 months of postnatal were 4.4 (Absent expression, A), 12.9 (A), 262.4 (Present expression, P), 1136.7 (P), 1617.5 (P) and 1128 (P),respectively. These results indicated that the expression of mouse Tpap wasn't detected on days 4 and 9, but was detected on days 18, 35, 54, and 6 months of mouse testis in our Affymetrix chip analysis. By combination with the RT-PCR analysis of mouse Tpap, we observed mouse Tpap began to express at the age of day 18 in mouse. Tpap is an age-dependent gene in mouse testis.The expression of Tpap corresponds to the appearance of spermatids of mice and indicates that Tpap may have an important role in male mammalian spermatogenesis.