Table3_Multiomic analysis revealed the regulatory role of the KRT14 gene in eggshell quality.XLSX

Background: Eggshell strength and thickness are critical factors in reducing the egg breaking rate and preventing economic losses. The calcite biomineralization process is very important for eggshell quality. Therefore, we employed transcriptional sequencing and proteomics to investigate the differences between the uteruses of laying hens with high- and low-breaking-strength shells.
Results: A total of 1,028 differentially expressed genes (DEGs) and 270 differentially expressed proteins (DEPs) were identified. The analysis results of GO terms and KEGG pathways showed that most of the DEGs and DEPs were enriched in vital pathways related to processes such as calcium metabolism, hormone and amino acid biosynthesis, and cell proliferation and apoptosis. Several DEGs and DEPs that were coexpressed at mRNA and protein levels were verified. KRT14 (keratin-14) is a candidate gene (protein) obtained by multiple omics analysis due to the fold difference of KRT14 being the largest. After the overexpression of KRT14 in uterine epithelial cells, the expressions of OC116 (ovocleididin-116), CALB1 (calbindin 1), and BST1 (ADP-ribosyl cyclase 2) were found to be increased significantly, while the expression of OC17 (ovocleididin-17) was found to be decreased significantly.
Conclusion: In summary, this study confirms that during normal calcification, there are differences in ion transport between the uterus of hens producing high-breaking-strength eggshells and those producing low-breaking-strength eggshells, which may help elucidate the eggshell calcification process. The KRT14 gene may promote calcium metabolism and deposition of calcium carbonate in eggshells.

背景：蛋壳强度与厚度是降低破蛋率、减少经济损失的关键因素。方解石生物矿化过程对蛋壳品质至关重要。为此，本研究采用转录组测序（transcriptional sequencing）与蛋白质组学（proteomics）技术，探究高破壳强度蛋壳与低破壳强度蛋壳对应的产蛋鸡子宫组织的差异。
结果：共鉴定得到1028个差异表达基因（differentially expressed genes, DEGs）与270个差异表达蛋白（differentially expressed proteins, DEPs）。基因本体（Gene Ontology, GO）术语与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路富集分析结果显示，大部分差异表达基因与差异表达蛋白富集于钙代谢、激素与氨基酸生物合成、细胞增殖及凋亡等相关的关键通路。部分在mRNA与蛋白水平共表达的差异表达基因和差异表达蛋白得到了验证。由于角蛋白14（keratin-14, KRT14）的差异倍数最大，其为多组学分析筛选得到的候选基因（蛋白）。在子宫上皮细胞中过表达KRT14后，发现卵壳蛋白116（ovocleididin-116, OC116）、钙结合蛋白1（calbindin 1, CALB1）以及ADP核糖基环化酶2（ADP-ribosyl cyclase 2, BST1）的表达量显著上调，而卵壳蛋白17（ovocleididin-17, OC17）的表达量显著下调。
结论：综上，本研究证实，在正常钙化过程中，产高破壳强度蛋壳的产蛋鸡子宫与产低破壳强度蛋壳的产蛋鸡子宫之间，离子转运存在差异，该发现有助于阐明蛋壳钙化过程。KRT14基因可促进钙代谢与蛋壳中碳酸钙的沉积。