Bouma Gerrit J, Hart Geoffrey T, Washburn Linda L, Recknagel Andrew K, Eicher Eva M
The Jackson Laboratory, 600 Main St, Bar Harbor, ME 04609, USA.
Gene Expr Patterns. 2004 Nov;5(1):141-9. doi: 10.1016/j.modgep.2004.05.001.
New techniques are being applied to identify all the genes involved in mammalian gonad development and differentiation. As this list of genes increases, understanding the potential interactions between these genes will become increasingly difficult. We used a real time reverse transcription PCR (real time RTPCR) protocol to examine and compare the relative expression levels of 55 genes in individual mouse fetal gonads. Real time PCR analysis demonstrated that except for Sry, no differences in relative gene expression were detectable between XX and XY gonad/mesonephroi complexes at embryonic day (E)11.5. Following Sry peak expression at E11.5, a number of genes were expressed at significantly higher relative levels in E12-14 XY than XX gonads. Of six genes expressed at higher levels in E12.5-14 XX than XY gonads, three, Bmp2, Emx2, and Fgfr2, had not been reported previously. Our results caution that differential localization patterns observed with whole mount in situ hybridization techniques may not accurately reflect changes in transcript levels. We conclude that real time PCR is an efficient and powerful tool for studying multiple gene expression patterns during gonad development and differentiation, and can provide insight into gene interactions.
新技术正被应用于识别所有参与哺乳动物性腺发育和分化的基因。随着这个基因列表的增加，了解这些基因之间潜在的相互作用将变得越来越困难。我们使用实时逆转录PCR（实时RT-PCR）方案来检测和比较单个小鼠胎儿性腺中55个基因的相对表达水平。实时PCR分析表明，除了Sry基因外，在胚胎第11.5天（E11.5）时，XX和XY性腺/中肾复合体之间未检测到相对基因表达的差异。在E11.5时Sry基因表达达到峰值后，一些基因在E期12 - 14天的XY性腺中的相对表达水平明显高于XX性腺。在E期12.5 - 14天XX性腺中表达水平高于XY性腺的六个基因中，有三个基因，即Bmp2、Emx2和Fgfr2，此前尚未见报道。我们的结果提醒，通过全胚胎原位杂交技术观察到的差异定位模式可能无法准确反映转录水平的变化。我们得出结论，实时PCR是研究性腺发育和分化过程中多个基因表达模式的一种高效且强大的工具，并且能够为基因相互作用提供深入见解。
