基于转录组测序分析鹅毛早期形态发育差异机制

Mechanism of differences in early morphological development of goose feathers based on transcnptome sequencing

【目的】探究调控鹅胚胎期不同部位初级羽囊早期形态发生差异的关键候选基因,为进一步揭示影响鹅不同部位羽毛形态发生的调控机制提供理论参考。【方法】采集鹅胚胎期初级羽囊早期背部和腹部皮肤组织样品进行组织切片制备和H&E染色,观察初级羽囊在2个部位的形态学差异;选取处于初级羽囊早期背部和腹部皮肤组织样品各3个,纯化RNA并进行转录组测序和生物信息学分析,筛选得到2个组织的差异表达基因(DEGs)。【结果】观察皮肤组织形态学发现背部初级羽囊原基的直径大于腹部,而密度小于腹部;基于背部和腹部皮肤组织转录组筛选DEGs共702个,筛选阈值为|log2Fold Change|≥1(P<0.05),包括337个上调和365个下调DEGs;采用实时荧光定量PCR验证挑选的4个基因,其结果显示与转录组数据表达趋势一致;对DEGs进行KEGG通路分析,富集到WNT和TGF-β等信号通路,挑选的GREM1、SMOC1、SFRP4和CRABP1等4个基因,推测是调控鹅胚胎期背部和腹部初级羽囊原基形态差异的关键候选基因。【结论】通过对鹅胚胎期背部和腹部皮肤组织进行形态学和转录组分析表明,背部初级羽囊原基直径大于腹部,而密度小于腹部,且有效筛选出影响初级羽囊原基形态差异的关键基因,可为进一步阐述影响鹅不同部位羽毛形态差异的分子机制和调控网络打下基础。

【Objective】The purpose of this paper was to explore the key candidate genes that regulated the early mor-phological differences of the primary feather follicle in different parts of goose during embryonic period,so as to provide theoretical reference for further revealing the regulation mechanism of feather morphogenesis in different parts of goose.【Method】In this study,dorsal skin and ventral skin tissue samples of goose during embryonic period in the early stage of primary feather follicle were collected for histological section preparation and H&E staining,the morphological differences between the two parts of the primary feather follicle were observed.Three dorsal skin tissue samples and three ventral skin tissue samples from goose during embryonic period in the early stage of primary feather follicle were collected.The RNA was purified and subjected to transcriptome sequencing and bioinformatics analysis,and differentially expressed genes(DEGs)of 2 tissues were screened.【Result】Observation of skin tissue morphology revealed that the diameter of the dorsal primary feather follicle primordium was longer than that of ventral part,but the density was lower than that of ventral part;a total of 702 DEGs were screened based on the transcriptomes of dorsal and ventral skin tissues,the screening threshold was|log2 Fold Change|≥1(P<0.05),including 337 up-regulated and 365 down-regulated DEGs;real-time fluo-rescence quantification(qRT-PCR)was used to verify the four genes selected,which was consistent with the expression trend of the transcriptome data according to the result;KEGG pathway analysis was performed on the DEGs,and WNT and TGF-βsignaling pathways were enriched.The 4 genes selected,GREM1,SMOC1,SFRP4 and CRABP1,were pre-sumed to be the key candidate genes regulating the morphological differences of dorsal and ventral primary feather follicle primordium in goose during embryonic period.【Conclusion】The histology and transcriptome analysis of dorsal and ven-tral skin tissues from goose at embryonic stage shows that the diameter of dorsal primary feather follicle primordium is longer than that of ventral part,but the density is lower than that of ventral part.The key genes affecting the morphological differences of the primary feather follicle primordium are effectively screened,which can provide basis for further eluci-dating the molecular mechanisms and regulatory networks affecting the morphological differences of feathers in different parts of goose.