Mabrouk Ichraf, Zhou Yuxuan, Wang Sihui, Song Yupu, Fu Xianou, Xu Xiaohui, Liu Tuoya, Wang Yudong, Feng Ziqiang, Fu Jinhong, Ma Jingyun, Zhuang Fangming, Cao Heng, Jin Honglei, Wang Jingbo, Sun Yongfeng
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China.
Key Laboratory of Animal Production, Product Quality and Security, Jilin Agricultural University, Ministry of Education, Changchun 130118, China.
Animals (Basel). 2022 Aug 17;12(16):2099. doi: 10.3390/ani12162099.
Skin and feather follicle development are essential processes for goose embryonic growth. Transcriptome and next-generation sequencing (NGS) network analyses were performed to improve the genome of Zhedong White goose and discover the critical genes, miRNAs, and pathways involved in goose skin and feather follicle morphogenesis. Sequencing output generated 6,002,591,668 to 8,675,720,319 clean reads from fifteen libraries. There were 1234, 3024, 4416, and 5326 different genes showing differential expression in four stages, E10 vs. E13, E10 vs. E18, E10 vs. E23, and E10 vs. E28, respectively. The differentially expressed genes (DEGs) were found to be implicated in multiple biological processes and pathways associated with feather growth and development, such as the Wnt signaling pathway, cell adhesion molecules, ECM-receptor interaction signaling pathways, and cell cycle and DNA replication pathways, according to functional analysis. In total, 8276 DEGs were assembled into twenty gene profiles with diverse expression patterns. The reliability of transcriptome results was verified by real-time quantitative PCR by selecting seven DEGs and five miRNAs. The localization of forkhead box O3 (FOXO3), connective tissue growth factor (CTGF), protein parched homolog1 (PTCH1), and miR-144-y by in situ hybridization showed spatial-temporal expression patterns and that FOXO3 and miR-144-y have an antagonistic targeting relationship. The correlation coefficient of FOXO3 and miR-144-y was -0.948, showing a strong negative correlation. Dual-luciferase reporter assay results demonstrated that miR-144-y could bind to the expected location to suppress the expression of FOXO3, which supports that there is a targeting relationship between them. The detections in this report will provide critical insight into the complex molecular mechanisms and breeding practices underlying the developmental characteristics of skin and feather follicles in Zhedong white geese.
皮肤和毛囊发育是鹅胚胎生长的重要过程。进行了转录组和下一代测序(NGS)网络分析,以完善浙东白鹅的基因组,并发现参与鹅皮肤和毛囊形态发生的关键基因、miRNA和信号通路。测序输出从15个文库中产生了6,002,591,668至8,675,720,319条clean reads。分别在E10与E13、E10与E18、E10与E23以及E10与E28这四个阶段中,有1234、3024、4416和5326个不同基因呈现差异表达。根据功能分析,发现差异表达基因(DEGs)涉及多个与羽毛生长发育相关的生物学过程和信号通路,如Wnt信号通路、细胞粘附分子、细胞外基质-受体相互作用信号通路以及细胞周期和DNA复制通路。总共8276个DEGs被组装成20个具有不同表达模式的基因图谱。通过选择7个DEGs和5个miRNA,利用实时定量PCR验证了转录组结果的可靠性。通过原位杂交对叉头框O3(FOXO3)、结缔组织生长因子(CTGF)、蛋白质patched同源物1(PTCH1)和miR-144-y进行定位,显示出时空表达模式,并且FOXO3和miR-144-y具有拮抗靶向关系。FOXO3与miR-144-y的相关系数为-0.948,呈现出强负相关。双荧光素酶报告基因检测结果表明,miR-144-y可以结合到预期位置以抑制FOXO3的表达,这支持了它们之间存在靶向关系。本报告中的检测将为浙东白鹅皮肤和毛囊发育特征背后的复杂分子机制及育种实践提供关键见解。