CAJ | 학술논문

孤雌胚胎的发育率比体内体外生成胚胎的发育率要慢，为研究小鼠孤雌胚、体外培养胚 H3K9乙酰化(H3K9ac)模式与体内自然胚之间的差异、曲古抑菌素 A(Trichostatin，TSA)对孤雌胚 H3K9乙酰化模式的影响及表观遗传模式对孤雌胚、体外培养胚发育的影响，文章采用间接免疫荧光法对小鼠植入前各时期孤雌胚、体外培养胚及体内自然胚基因组组蛋白的H3K9乙酰化水平进行检测。结果显示，植入前各时期孤雌胚H3K9乙酰化模式与体内组变化趋势基本一致，但平均荧光强度较体内组普遍偏高；经 TSA 处理后孤雌胚 H3K9乙酰化水平有所提高，原核期至8-细胞期差异显著(P<0.05)。体外培养胚H3K9乙酰化荧光强度与体内组变化趋势也基本一致，但平均荧光强度较体内组普遍偏低。以上结果表明，小鼠孤雌胚 H3K9乙酰化水平高于体内胚，使植入前胚胎发育过程中本应沉默的基因启动子发生超乙酰化，进而抑制胚胎发育，这可能是造成孤雌胚胎发育能力较差的重要原因之一；TSA处理可以部分弥补体外培养环境对胚胎发育带来的伤害，但 TSA提高孤雌胚的发育能力可能并不完全是通过改变H3K9乙酰化水平来实现的。
고자배태적발육솔비체내체외생성배태적발육솔요만，위연구소서고자배、체외배양배 H3K9을선화(H3K9ac)모식여체내자연배지간적차이、곡고억균소 A(Trichostatin，TSA)대고자배 H3K9을선화모식적영향급표관유전모식대고자배、체외배양배발육적영향，문장채용간접면역형광법대소서식입전각시기고자배、체외배양배급체내자연배기인조조단백적H3K9을선화수평진행검측。결과현시，식입전각시기고자배H3K9을선화모식여체내조변화추세기본일치，단평균형광강도교체내조보편편고；경 TSA 처리후고자배 H3K9을선화수평유소제고，원핵기지8-세포기차이현저(P<0.05)。체외배양배H3K9을선화형광강도여체내조변화추세야기본일치，단평균형광강도교체내조보편편저。이상결과표명，소서고자배 H3K9을선화수평고우체내배，사식입전배태발육과정중본응침묵적기인계동자발생초을선화，진이억제배태발육，저가능시조성고자배태발육능력교차적중요원인지일；TSA처리가이부분미보체외배양배경대배태발육대래적상해，단 TSA제고고자배적발육능력가능병불완전시통과개변H3K9을선화수평래실현적。
The developmental rate of parthenogenetic embryos is slower than that of embryos generated in vitro and in vivo. To detect the effects of epigenetic modification on embryo development, we compared the H3K9 acetyla-tion level in these three types of embryos as well as parthenogenetic embryos treated with a histone deacetylase in-hibitor trichostatin (TSA) by indirect immunofluorescence. Our results showed that fluctuations in the level of acety-lated H3K9 detected during embryo development are similar among different types of mouse embryos. However, the level of H3K9 acetylation in parthenogenetic embryos is significantly higher while the level in embryos generated in vitro is lower when compared with that in embryos derived from in vivo. Treatment of parthenogenetic embryos with TSA increases the developmental rate but further elevates the level of H3K9 acetylation, especially from pronuclear to 8-cell stages. These results suggest that the promoters of genes that should be silenced during pre-implantation embryo development may be hyperacetylated in parthenogenetic embryos which inhibit normal embryo development. However, the positive effect of TSA on embryo development is not through altering the H3K9 acetylation level.