Developmental Changes in Expression of Genes Involved in Regulation of Apoptosis in the Bovine Preimplantation Embryo
Developmental Changes in Expression of Genes Involved in Regulation of Apoptosis in the Bovine Preimplantation Embryo
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DOI：
10.1095/biolreprod.110.086249
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发表时间：
2011-01-01
影响因子：
3.6
通讯作者：
Hansen, Peter J.
中科院分区：
文献类型：
--
作者：
Fear, Justin M.;Hansen, Peter J.
早期牛植入前胚胎对促凋亡信号具有抗性，直到大约8- 16细胞阶段。我们假设，与>= 16细胞的胚胎相比，2-细胞胚胎具有更高量的抗凋亡蛋白和更低量的促凋亡蛋白。抗凋亡基因BCL 2和HSPA 1A的mRNA的稳态浓度在MII卵母细胞、2-细胞胚胎、与>= 16细胞胚胎相比，用α鹅膏蕈碱处理的2-细胞胚胎中促凋亡基因BAD的稳态mRNA浓度在>= 16细胞胚胎中增加。发育阶段对BCL 2L 1、DFFA或BAX的稳态mRNA浓度没有显著影响。发现BCL 2在2-细胞胚胎中的相对浓度大于在>= 16细胞胚胎中的相对浓度。这些结果通过Western印迹得到证实，通过免疫荧光检测到的免疫反应性BAX的相对量在2-细胞胚胎中低于在>= 16细胞胚胎中。高分子量（46 kDa）形式的BAX在≥ 16细胞胚胎中最高，在2细胞胚胎中居中，在MII卵母细胞中最低。发育阶段对免疫反应性BCL 2L 1、HSPA 1A或BAD的相对量没有影响，从第0天至第5天或第4天至第1天用α鹅膏蕈碱处理胚胎）授精后在受精后第5天用促凋亡信号C-2神经酰胺处理后降低了II组半胱天冬酶和末端脱氧核苷酸转移酶dUTP缺口末端标记的活化因此，BAX或其他蛋白质的转录是获得细胞凋亡能力所必需的。结果支持了BCL 2家族成员数量的变化对于抑制2-HT细胞中的细胞凋亡是重要的这一观点。细胞胚胎和建立细胞凋亡的能力以后的发展
The early bovine preimplantation embryo is resistant to proapoptotic signals until around the 8- to 16 cell stage We hypothesized that 2-cell embryos have higher amounts of antiapoptotic proteins and lower amounts of proapoptotic proteins when compared to embryos >= 16 cells Steady-state concentrations of mRNA for the antiapoptotic genes BCL2 and HSPA1A were higher for MII oocytes, 2-cell embryos, and 2-cell embryos treated with alpha amanitin as compared to >= 16-cell embryos Steady state concentrations of mRNA for the proapoptotic gene BAD increased in embryos >= 16 cells There was no significant effect of stage of development on steady-state mRNA concentrations of BCL2L1, DFFA, or BAX Using immunohistochemistry, it was found that BCL2 was present in greater relative concentrations for 2-cell embryos than for embryos >= 16 cells These results were confirmed by Western blotting Relative amounts of immunoreactive BAX detected by immunofluorescence were lower for 2-cell embryos than for embryos >= 16 cells Using Western blotting, a high molecular weight (46 kDa) form of BAX was highest in >= 16-cell embryos, intermediate in 2-cell embryos, and lowest in MII oocytes There were no effects of stage of development on relative amounts of immunoreactive BCL2L1, HSPA1A, or BAD, as determined by immunofluorescence Treatment of embryos with alpha amanitin from Day 0 to Day 5 or Day 4 to Day 1) after insemination reduced activation of group II caspases and terminal deoxynucleotidyl transferase dUTP nick end labeling after treatment with the proapoptotic signal C-2 ceramide at Day 5 after fertilization Thus, transcription of BAX or other proteins is required for acquisition of the capacity for apoptosis Results support the Idea that changes in amounts of BCL2 family members are important for the inhibition of apoptosis in the 2-cell embryo and in the establishment of the capacity for apoptosis later in development