利用原位杂交和本实验室建立的猪HSPs mRNA一步法实时荧光定量RT—PCR技术平台等方法,研究长途运输对猪肾脏中HSP7。及HSP90 BmRNA转录水平及在组织细胞中定位的影响。经1,2,4,6和10h的运输应激后,肾组织中HSP70 mRNA的转录水平随着运输应激时间的延长一直呈现出上升的趋势,运输应激持续到10h时,HSP70 mRNA的转录水平升到最高,为对照组的14.2倍;HSP90 mRNA的转录水平在应激初期(1—2h)急剧升高(P〈0.01),到运输应激持续到6h后,升高不再显著。HSPs mRNA在肾组织细胞中的定位无肉眼可见的随着应激时间的延长而发生相应变化。HSP70 mRNA和HSP90 mRNA在肾小球毛细血管内皮细胞的胞核以及肾小管上皮细胞中呈阳性分布。运输应激可以导致HSPs mRNA转录水平的变化,并且对不同家族的HSPs mRNA转录水平的影响不同;HSP70 mRNA的转录水平随着应激时间的延长一直呈现出上升的趋势,有望成为猪运输应激的判定指标之一。
In situ hybridization and one - step fluorescence quantitative methods were used to invest the changes of HSPs mRNA distribution and transcription level in kidney of transport stressed pigs. The level of HSP70 mRNA transcription increased continuously from the beginning of transportation. The inductions of HSP70 mRNA transcription in kidney of 10 - hour - transport stressed pigs were 14.2 times higher than those of the un- transport stressed pigs(P 〈0.01 ). However, the transcription levels of HSPgo mRNA increased only at the beginning of transport stress. HSP70 and HSPgo mRNA expressed preferentially in some kidney tubules. Transport stress can affect the transcription of HSPs mRNA, and the effect is different between HSPs families. Furthermore, the concentration of HSPTo mRNA increased continuously from the beginning of transport stress, might be a hallmark to determine stress.