摘要
将40羽220日龄ISA蛋鸡分为5组,于产蛋后0、2、4.5、8、16h断头处死,采集蛋壳腺组织,运用Real-time PCR和Western-blot方法检测产蛋循环过程中瞬时性受体单位香草精受体6（Transient receptor potential vanilloid receptor 6,TRPV6）、钙结合蛋白（calbindin D28K,CaBP-D28K）、质膜钙离子ATP酶1b（plasma membrane Ca2＋-ATPase 1b,PMCA 1b）mRNA和蛋白浓度的动态变化。结果显示,卵子未进入蛋壳腺前（产蛋后0~4.5h）,蛋壳腺内TRPV6、CaBP-D28K和PMCA 1bmRNA表达水平较低,随后表达量逐渐升高,在蛋壳钙化过程中达到最大值（产蛋后16h）,与0h相比,TRPV6和CaBP-D28KmRNA表达差异均极显著（P〈0.01）;另外,产蛋循环过程中,TRPV6、CaBP-D28K和PMCA 1b蛋白浓度变化与mRNA变化一致,产蛋后16h到达最大值,其中CaBP-D28K蛋白浓度与0h相比,差异显著（P〈0.05）。结果表明,产蛋循环可调控蛋壳腺内TRPV6、CaBP-D28K和PMCA 1b的表达,并提示钙离子跨细胞转运途径在钙离子进入蛋壳腺形成蛋壳过程中发挥重要作用。
The aim of this study is to investigate the effect of active Ca2＋transcellular transport on the eggshell calcification.Forty ISA laying hens at the peak stage（220days old）were assigned to five reproductive stages for sampling.The samples of egg shell gland（ESG）were obtained by decapitated at 0,2,4.5,8and 16 hafter post-oviposition,respectively.Then,quantitation of mRNA level and protein concentration of TRPV6,CaBP-D28 Kand PMCA 1bat different stages in the ESG were carried out by real-time PCR and Western-blot analysis,respectively.The results are as follows：the expression levels of TRPV6,CaBP-D28 Kand PMCA 1bmRNA in the ESG retained very low until the egg movement into the shell gland（0-4.5hafter ovulation）,then significantly increased at 16 hduring eggshell calcification.In addition,TRPV6,and CaBP-D28 Kindicated significant statistical difference（P0.01）,respectively.Furthermore,western blotting showed that the expression of TRPV6 and PMCA 1breached a maximum at 16 hafter ovulation,but the statistical difference was not significant.The change of CaBP-D28 Kexpression was very similar to that of TRPV6,but the concentration was significantly higher at 16 hthan that at 0hafter ovulation（P0.05）.In conclusion,the expression of TRPV6,CaBP-D28 Kand PMCA 1bin the ESG was regulated by the oviposition cycle,suggesting that active Ca2＋transcellular transport exerted significant effects in calcium delivering in the ESG.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2014年第9期1534-1539,共6页
Chinese Journal of Veterinary Science
基金
上海市科技兴农重点攻关资助项目(沪农科攻字(2013)第3-8号)
作者简介
翟晓虎（1981-），男，讲师，博士。
通讯作者，E—mail：yangjunhua303@126．com
