Wu Zhihong, Wang Yu, Han Wenjing, Yang Kun, Hai Erhan, Ma Rong, Di Zhengyang, Shang Fangzheng, Su Rui, Wang Ruijun, Wang Zhiying, Zhang Yanjun, Li Jinquan
College of Animal Science, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia Autonomous Region 010018, China.
College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia Autonomous Region 010018, China.
Arch Anim Breed. 2020 Dec 10;63(2):461-470. doi: 10.5194/aab-63-461-2020. eCollection 2020.
This study is focused on the detection of ectodysplasin A (EDA) and ectodysplasin A receptor (EDAR) mRNA expression levels and protein positions in seven stages of cashmere goat fetus development (45, 55, 65, 75 95, 115, and 135 d), with the main goal of investigating the effect of EDA and EDAR on genes related to hair follicle development. Quantitative real-time polymerase chain reaction (RT-qPCR) was used to measure EDA and EDAR expression levels in seven stages of cashmere goat fetus development. Immunohistochemistry (IHC) was used to locate EDA and EDAR in the critical stage of fetal hair follicle development (45-135 d). EDA and EDAR expression in fetal fibroblasts and epithelial cells was interfered with by short hairpin RNA (sh-RNA). The results indicated that EDA and EDAR were both expressed in the skin tissue in the seven cashmere goat embryo stages. Moreover, EDA and EDAR play an important role in the formation of embryonic placode (Pc). After interfering with EDA and EDAR, the expression of BMP2, BMP4, noggin, -catenin, TGF- 2, Wnt-10b, and NOTCH1 in fibroblasts and epithelial cells changed significantly. This study provides a theoretical and experimental basis for further studying the molecular regulation mechanism of hair follicle development.
本研究聚焦于检测绒山羊胎儿发育七个阶段（45、55、65、75、95、115和135天）中外胚层发育不良蛋白A（EDA）和外胚层发育不良蛋白A受体（EDAR）的mRNA表达水平及蛋白定位，主要目的是研究EDA和EDAR对毛囊发育相关基因的影响。采用定量实时聚合酶链反应（RT-qPCR）测定绒山羊胎儿发育七个阶段中EDA和EDAR的表达水平。利用免疫组织化学（IHC）在胎儿毛囊发育的关键阶段（45 - 135天）定位EDA和EDAR。通过短发夹RNA（sh-RNA）干扰胎儿成纤维细胞和上皮细胞中EDA和EDAR的表达。结果表明，EDA和EDAR在绒山羊胚胎的七个阶段的皮肤组织中均有表达。此外，EDA和EDAR在胚胎基板（Pc）的形成中起重要作用。干扰EDA和EDAR后，成纤维细胞和上皮细胞中BMP2、BMP4、头蛋白、β-连环蛋白、TGF-β2、Wnt-10b和NOTCH1的表达发生显著变化。本研究为进一步研究毛囊发育的分子调控机制提供了理论和实验依据。
