肉鸡胫骨软骨发育不良早期钙黏蛋白1(CDH1)差异表达研究

The Differential Expression of Cadherin 1(CDH1) in the Growth Plate of Tibial Dyschondroplasia in Broiler Chickens at the Early Stage

为研究福美双诱发肉鸡胫骨软骨发育不良(Tibial Dyschondroplasia,TD)早期钙黏蛋白1(CDH1)的差异表达,基础日粮中添加福美双,在试验第1、2、6天,对试验鸡进行剖杀,迅速采取胫骨生长板放入4%多聚甲醛溶液于4℃固定,做CDH1免疫组化分析;提取对照组和饲喂福美双组的生长板总RNA,采用Real-time PCR对CDH1基因进行差异表达验证。结果钙黏蛋白1(CDH1/E-cadherin)基因在TD生长板表达上调,在对照和TD软骨生长板,其蛋白合成主要在前肥大、肥大区软骨细胞质,增殖区软骨细胞无表达,钙化区软骨细胞表达少,在饲喂福美双第1、2、6天其表达增加与定量PCR变化结果相一致,且在饲喂福美双第2、6天呈明显高表达。结果表明CDH1参与细胞黏附、血管入侵及调节Wnt/β-cat的信号传递,和其它分子共同调节软骨内骨化。

The aim of this study was to explore the differential expression of cadherin 1（CDH1） in the growth plate of broiler chickens with tibial dyschondroplasia（TD） induced by thiram at the early stage.Chickens were dissected and growth plates were fixed in 4% paraform respectively in 4 ℃ at days 1,2 and 6.RNA was extracted from the growth plates of control and thiram-fed chickens.Then differential expression of CDH1 was identified by real-time PCR and immunohistology.The results showed that expression of CDH1 was up-regulated on the growth plate of thiram-diet fed group.CDH1 protein synthesis mainly lied in cytoplasm of the prehypertrophic and hypertrophic chondrocyte in control or TD growth plates.No expression was discovered at proliferative zone and low expression was detected at calcification zone.Remarkably increased CDH1 was detected in TD growth plates at days 1,2 and 6 after feeding thiram diet,which mirrored with their mRNA differential expression,and highly increased CDH1 staining was clearly seen at days 2 and 6 respectively.It was suggested that CDH1 was associated with cell adhesion,blood vessel invasion and it also regulated Wnt/β-cat signal transmission with other regulator among endochondral bone formation.