利用设计合成的地高辛标记的黑腹果蝇(Drosophila melanogaster)hsp70DNA探针,整体原位杂交检测热休克反应中,hsp70mRNA在胚胎发育各阶段的表达情况.实验发现,在正常温度下黑腹果蝇的胚胎检测不到hsp70mRNA存在.在热激的果蝇胚胎中,可检测到hsp70mKNA的广泛存在.hsp70mRNA在热激的胚胎中,除早期细胞化囊胚的极细胞外,其余细胞均可检测到hsp70mRNA的均一分布.随着胚胎发育,hsp70mRNA仍广泛分布在各细胞中,晚期胚胎中神经细胞hsp70mKNA比其余细胞的表达水平高,说明神经细胞具有更高的热诱导活性,在热激反应中hsp70mRNA的表达水平更高.
This paper reported the results of hsp70 mRNA detected in heat shock and no heat shock Drosophila melanogaster embryos via in situ hybridization. The digoxigenin-labelled DNA probes were synthesized using PCR. The antisense probe was used to detect hsp70 mRNA in heat-shocked embryos and non heat shock embryo. The sense probe was used as negative control to detect hspTO mRNA in heat shocked embryos. In-sire hybridization was used to detect hsp70 mRNA distribution in these two treatment embryos. We found that hsp70 mRNA could not detected in non heat shock embryos. But in the heat shocked embryos, the distribution of hsp70 mRNA disphyed a complex expression pattern. All cells in early embryos showed the existence of hsp70 mRNA except the pole cells. With the embryo development, hsp70 mRNA also presented in all cells, especially abundant in the neuronal cells. It seemed like neuronal cells had a higher heat-sensitivity than other soma cells, The expressing of hsp70 mRNA was easily induced by heat shock in these cells.