Table4_Effects of exogenous melatonin on expressional differences of immune-related genes in cashmere goats.xlsx

The interplay between melatonin and immune system is well recognized in humans. The true integration of research on cashmere goat is still far from clear, especially for cashmere goat maintained in wool and cashmere growth. In this study, we applied various approaches to identify the complex regulated network between the immune-related genes and transcription factors (TFs) and to explore the relationship between melatonin and gene expression in cashmere goats. In total, 1,599 and 1756 immune-related genes were found in the blood and skin of cashmere goats, respectively, and 24 differentially expressed immune-related GO terms were highly expressed in blood after melatonin implantation. We studied the melatonin-dependent networks between the TFs and immune-related genes in cashmere goat. The 3 major regulatory networks were interconnected through TFs. The TFs, such as PHF5A, REXO4, STRAP, JUNB, GATAD2A, ZNF710, and VDR, were also expressed in the blood and skin tissue of cashmere goat. In addition, most genes in these networks, such as VDR, JUNB, and Trib3, were involved in WNT pathway, which is related to cashmere wool growth regulation. On the network basis, we developed a knockout mouse model to identify the network interaction. We observed that 8 high-sulfur protein genes, 12 keratin (KRT) genes, and 19 keratin associated protein (KRTAP) genes related to the growth of cashmere wool were almost not expressed in Trib3−/− rat skin. Our results suggested that the expression of genes related to wool and cashmere growth may be regulated by the interaction network between genes affected by melatonin and immune-related genes. In summary, we outlined some particularly promising ways for future research on immune-related genes of cashmere goats and the role of melatonin in wool and cashmere growth.

褪黑素（melatonin）与免疫系统之间的相互作用在人类中已得到广泛认可。然而针对绒山羊的相关研究的真正整合仍远未明晰，尤其是在维持羊毛与羊绒生长的研究方向上。本研究采用多种方法，解析了免疫相关基因与转录因子（transcription factors, TFs）之间的复杂调控网络，并探究了褪黑素与绒山羊基因表达之间的关联。共计分别在绒山羊的血液与皮肤组织中鉴定出1599个和1756个免疫相关基因；在褪黑素植入后，血液中有24个差异表达的免疫相关基因本体（Gene Ontology, GO）条目呈现高表达水平。本研究针对绒山羊中依赖褪黑素的转录因子与免疫相关基因调控网络展开分析，发现3个主要调控网络通过转录因子实现相互连接。诸如PHF5A、REXO4、STRAP、JUNB、GATAD2A、ZNF710及VDR等转录因子，在绒山羊的血液与皮肤组织中均有表达。此外，这些网络中的多数基因（如VDR、JUNB及Trib3）均参与了与羊绒生长调控相关的WNT信号通路。基于该调控网络，本研究构建了基因敲除小鼠模型以解析网络间的相互作用。我们观察到，在Trib3基因敲除大鼠的皮肤组织中，与羊绒生长相关的8个高硫蛋白基因、12个角蛋白（keratin, KRT）基因及19个角蛋白关联蛋白（keratin associated protein, KRTAP）基因几乎均未表达。本研究结果表明，与羊毛及羊绒生长相关的基因表达，可能通过受褪黑素调控的基因与免疫相关基因之间的相互作用网络实现调控。综上，本研究为绒山羊免疫相关基因的后续研究，以及褪黑素在羊毛与羊绒生长中的作用机制探索，指明了若干极具前景的方向。