雏鹅冷应激反应中HPA、HPT轴应激激素相关基因表达调控规律

Analysis on Dynamic Expression of Gene mRNA Related with Hormone in HPA and HPT Axis

以7日龄皖西白鹅为试验对象,研究于（12±1）℃条件下进行急性冷应激处理时对HPA、HPT轴的影响。利用荧光定量逆转录PCR（FQ-RT-PCR）技术动态研究冷应激反应中应激激素相关基因TRH、TSH、CRH、ACTH基因mRNA的变化规律。研究结果表明,下丘脑TRH基因mRNA转录量在冷应激0.5 h3、h、6 h9、h1、8 h与应激结束后1 h6、h和12 h显著升高,其他时间内均下降到室温时的转录水平。垂体TSH基因mRNA转录量在冷应激1.5 h到12 h期间显著升高,应激12 h时转录水平达到最高点,而其他应激时间均降低到应激前的水平,应激结束后1 h到6 h期间垂体TSH基因mRNA转录水平也被显著升高,冷处理结束后9~24 h已恢复到应激前的正常水平。下丘脑CRH基因mRNA转录量在冷应激3h、9 h1、8 h时与应激结束后0.5 h、1 h和9 h显著升高,其他时间点降到应激前的正常水平。肾上腺ACTH基因mRNA转录水平在冷应激1.5~9 h、18 h时显著升高,其他时间点均迅速下降到应激前水平;冷处理结束后0.5 h和3 h,由于又突然受到热的室温应激,ACTH基因mRNA转录量又显著升高,其他时间点降到应激前的正常水平。下丘脑TRH基因mRNA表达呈现波动性变化趋势,但表达量均不超过1.5,说明下丘脑TRH基因是负调控的。下丘脑CRH、垂体TSH和肾上腺ACTH基因mRNA的表达量分别在2.27~7.76、1.75~6.17、5.5~14.83范围内,总体呈上升趋势,说明TSH、CRH和ACTH基因均是冷应激反应时的正调控基因。

Wanxi White geese were used to study the effects on the dynamic expression of TRH,TSH,CRH and ACTH mRNA in HPA and HPT axis under cold stress（12±1）℃ by FQ-RT-PCR.Hypothalamic TRH mRNA transcription level increased significantly at 0.5 h,3 h,6 h,9 h and 18 h during cold stress.It increased again after cold exposure in 1 h,6 h and 12 h,and decreased to normal level in the rest time.Pituitary TSH mRNA transcription level was increased significantly from 1.5 h to 12 h,moreover,it reached the highest level at 12 h,then it declined to normal level.It increased again after cold exposure from 1 h to 6h and was decreased to normal level after 6h.Hypothalamic CRH mRNA transcription level increased significantly at 3 h,9 h,18 h during cold stress and at 1 h,6 h and 12 h after cold exposure.It was down-regulated to normal level at the rest time.Adrenal gland ACTH gene mRNA transcription level increased significantly from 1.5 h to 9 h and at 18 h during cold stress,and increased again after cold exposure at 0.5 h and 3 h.Hypothalamic TRH mRNA transcription level had fluctuation trend and it＇s quantity was not over 1.5,which indicated that hypothalamic TRH was the down-regulation gene in cold stress reaction.The transcription quantity of TSH,CRH and ACTH was 2.27-7.76、1.75-6.17、5.5-14.83 and had ascend trend,which showed that they were the up-regulation gene in cold stress reaction.The results demonstrated that TRH,TSH,CRH and ACTH gene could be probably used as the molecular biomarker of animal cold stress.