microRNA-let-7家族在约克夏猪甲状腺生长发育过程中的表达分析  

Expression Analysis of let- 7 microRNAs in Yorkshire Pig's(Sus scrofa)Thyroid at Different Growth Stages

作　　者：王琪[1] 王云霞[1] 王颖[2] 徐宁迎[1] 

机构地区：[1]浙江大学动物科学学院,杭州310058 [2]太原理工大学力学学院,太原030024

出　　处：《农业生物技术学报》2015年第3期337-343,共7页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31272424);太原理工大学引进人才项目(No.tyut-rc201334a)

摘　　要：miR-let-7广泛参与动物组织器官发育调控,是研究猪甲状腺发育的重要候选miRNAs。为了研究miR-let-7家族在约克夏猪(Sus scrofa)甲状腺生长发育过程中的作用,本实验一方面通过对不同发育阶段(胚龄105,出生后60、90、150日龄)的猪甲状腺组织切片进行苏木精-伊红染色(hematoxylin-eosin staining,HE),研究甲状腺发育形态学的变化;另一方面,采用原位杂交(in situ hybridization,ISH)和荧光定量PCR技术鉴定miR-let-7家族在不同生长阶段(胚龄105,出生后60、90、150日龄)约克夏猪甲状腺中的表达。HE染色结果发现,甲状腺滤泡随着日龄的增加逐渐增大。原位杂交结果显示,miR-let-7主要位于甲状腺的滤泡上皮细胞中,并且let-7的表达在出生后有先减少后增加的态势。通过荧光定量PCR结果发现,在生长期随着日龄的增加,let-7d-5p、let-7f和let-7g的相对表达量逐渐上升,let-7c、let-7d-3p、let-7e和let-7i的表达先升后降。研究结果提示,约克夏猪甲状腺miR-let-7的表达直接或间接影响甲状腺激素水平的变化,在甲状腺生长发育中发挥了重要的作用,为深入研究猪的生长发育提供了一个新的思路。The let-7 family of microRNAs （miR） which is widely involved in the regulation of development in animal tissues and organs, is an important candidate microRNAs of pig＇s thyroid development. In order to study the role of let-7 family in the development of Yorkshire pig＇s （Sus scrofa） thyroid gland, firstly, we stained pig＇s thyroid at different developmental stages （105 days in the embryonic period, 60, 90, 150 days in the youth period） with hematoxylin-eosin （HE） staining to discover thyroid morphological change; secondly, we used in situ hybridization （ISH） and fluorescence quantitative PCR （qRT-PCR） technique to identify the expression of let-7 microRNAs at different growth stages （105 days in the embryonic period, 60, 90, 150 days in the youth period）. The results showed that thyroid follicles gradually became bigger with age; the let-7 family was mainly located in the thyroid follicular epithelium; in the youth period the expression of let-7c, let- 7d-3p, let-Te, let-7i rose gradually, while the expression of let-Tc, let-7d-3p, let-7e, let-7 increased firstly and then decreased. Based on the above results, we predicted that the expression of let-7 had relation with the expression of thyroid hormone directly or indirectly, thereby playing an important role in thyroid growth. This study provides a new idea for further research on growth and development of pig.

关 键 词：miR-let-7 原位杂交(ISH) 甲状腺 甲状腺激素 

分 类 号：S858.31[农业科学—临床兽医学]