Chastant S, Christians E, Campion E, Renard J P
Unité de Biologie du Développement, INRA, Jouy-en-Josas, France.
Mol Reprod Dev. 1996 Aug;44(4):423-32. doi: 10.1002/(SICI)1098-2795(199608)44:4<423::AID-MRD1>3.0.CO;2-N.
HSP 70.1 is one of the first genes to be expressed in the mouse embryo at the time of zygotic genome activation. We studied the regulation of this gene, using a transgene associating HSP 70.1 promoter and the firefly luciferase reporter gene, which allows the precise quantification of HSP 70.1 level of expression on individual embryos. In the present work, we show first that the level of HSP 70.1 expression at the two-cell stage is significantly higher (around two-fold) in embryos whose maternal cytoplasm is from C3H strain than with BALB/c strain. We verified that this difference is not an artefact of the use of transgenic embryos, of the time of first cleavage, or of in vitro culture. This regulation of HSP 70.1 level of expression is controlled by strain-specific maternal modifiers and is independent of replication, syngamy, and mitosis. Following nuclear transfer, reactivation of HSP 70.1 is also subjected to the same epigenetic influence. Only the strain-of-origin of the recipient cytoplast modulates the level of HSP 70.1 reprogrammation; the origin of donor nucleus is not significant, demonstrating the reversibility of this strain effect. These results point out the importance of the quality of recipient cytoplast in the intensity of gene reprogrammation, which may be of importance for nuclear transfer efficiency.
热休克蛋白70.1(HSP 70.1)是合子基因组激活时在小鼠胚胎中最早表达的基因之一。我们使用一个将HSP 70.1启动子与萤火虫荧光素酶报告基因相连的转基因来研究该基因的调控,这使得能够精确量化单个胚胎中HSP 70.1的表达水平。在本研究中,我们首先发现,母本细胞质来自C3H品系的胚胎在二细胞阶段的HSP 70.1表达水平显著高于(约两倍)母本细胞质来自BALB/c品系的胚胎。我们证实,这种差异不是使用转基因胚胎、首次卵裂时间或体外培养造成的假象。HSP 70.1表达水平的这种调控由品系特异性的母本修饰因子控制,且与复制、受精和有丝分裂无关。核移植后,HSP 70.1的重新激活也受到相同的表观遗传影响。只有受体细胞质的来源品系调节HSP 70.1重编程的水平;供体细胞核的来源不显著,这证明了这种品系效应的可逆性。这些结果指出了受体细胞质质量在基因重编程强度中的重要性,这可能对核移植效率具有重要意义。