PTEN基因在鸡肝脏中的发育性变化

Developmental changes of the PTEN gene in chicken livers

为了研究抑癌基因PTEN在家禽肝脏发育过程中的表达变化,试验选取50枚海兰褐受精蛋,在孵化的第15,18,20天[E15、E18、E20(啄壳为准)]和出壳后第1,5,10,15天(D1、D5、D10、D15)各取6枚(只)鸡胚/雏鸡,用乙醚麻醉后颈椎脱臼致死,取肝脏并提取基因组,采用实时荧光定量PCR和免疫组织化学方法对PTEN基因在家禽肝脏发育过程中的表达变化进行研究。结果表明:PTEN基因相对表达量随着胚龄/日龄的增长而增加,但在啄壳当天,PTEN基因相对表达量会暂时下降。PTEN蛋白在出壳前期主要分布在部分肝血窦内皮细胞的细胞核和肝细胞的细胞核中,在细胞质中基本无表达;在出壳后,PTEN蛋白在肝细胞核和细胞浆内均有分布。E20,免疫阳性PTEN蛋白的表达量下降,和E15水平相当(P>0.05),在其他胚龄/日龄时均显著高于E15(P<0.05)。肝细胞核PTEN蛋白免疫阳性率在E18显著高于E15(P<0.05);在E20和D15,肝细胞核PTEN蛋白免疫阳性率急剧下降,均显著低于E18(P<0.05);在D5~D15,肝细胞核PTEN蛋白免疫阳性率逐渐增加,均显著高于E15(P<0.05)。PTEN基因的这种表达模式和鸡胚/雏鸡肝脏的发育模式基本一致,说明家禽PTEN基因在肝脏发育和代谢中发挥重要作用。

In order to study the expression changes of the tumor suppressor gene PTEN during the development of poultry liver,in this experiment,50 fertilized eggs Hy-line from brown chickens,and 6 chicken embryos/chicks from each egg were taken on the 15th,18th and 20th days of incubation[(E15),E18,E20(based on pecking)]and on the first 5th,10th and 15th days after shell extraction(D1,D5,D10,D15),respectively.Under ether anesthesia,the liver was extracted for genome extraction after death by cervical dislocation.The changes of PTEN gene expression during poultry liver development was studied by real-time fluorescence quantitative PCR and immunohistochemistry.The results showed that the relative expression of PTEN gene increased with the increase of embryonic age/day of age,but the relative expression of PTEN gene decreased temporarily on the day of shell pecking.The PTEN protein was mainly distributed in the nucleus of some liver sinusoidal endothelial cells and the nucleus of liver cells during the prophase of shell formation,and almost no expression was found in the cytoplasm.After hatching,the PTEN protein was distributed in the nucleus and cytoplasm of hepatocytes.The average optical density of PTEN protein was decreased at E20,which was similar to that at E15(P>0.05),and the average absorbance value of PTEN protein immunopositive at other embryo ages/day was significantly higher than that at E15(P<0.05).The immunopositive rate of PTEN protein in the liver nucleus at E18 was significantly higher than that at E15(P<0.05).At E20 and D1,the immunopositive rate of PTEN protein in the liver nuclei decreased sharply,which was significantly lower than that at E18(P<0.05).The immunopositive rate of PTEN protein in the liver nucleus increased gradually from D5 to D15,and was significantly higher than that at E15(P<0.05).This pattern of PTEN expression was consistent with the developmental pattern of chick embryo or chick liver.These results indicated that PTEN plays an important role in liver development and metabolism in poultry.