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. 2022 Sep 22:13:927670.
doi: 10.3389/fgene.2022.927670. eCollection 2022.

Multiomic analysis revealed the regulatory role of the KRT14 gene in eggshell quality

Yan Wu  1   2 , Yanyan Sun  3 , Hao Zhang  1 , Hongwei Xiao  1 , Ailuan Pan  1 , Jie Shen  1 , Yuejin Pu  1 , Zhenhua Liang  1 , Jinping Du  1 , Jinsong Pi  1
Affiliations

Multiomic analysis revealed the regulatory role of the KRT14 gene in eggshell quality

Yan Wu et al. Front Genet. .

Abstract

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.

Keywords: KRT14 gene; eggshell; laying hen; proteome; transcriptome.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Differentially expressed genes (DEGs) identified for the uterus between HE and LE hens. (A). Volcano plot of DEGs between HE and LE hens; (B). GO analysis result of DEGs between HE and LE hens; (C). KEGG analysis result of DEGs between HE and LE hens.
FIGURE 2
FIGURE 2
Differentially expressed proteins (DEGs) identified for the uterus between HE and LE hens. (A) GO analysis result of DEPs between HE and LE hens; (B) top 20 results of KEGG analysis of DEPs between HE and LE hens; (C) protein–protein interaction network of the DEPs.
FIGURE 3
FIGURE 3
Integration and analysis of transcriptome and proteome between HE and LE groups. (A). DEGs, DEPs, and Co-DEGs-DEPs between HE and LE groups; (B). Venn diagram for DEGs and DEPs between HE and LE groups; (C). Corr_plot of RNA-seq and TMT between HE and LE groups; (D). Share GO analysis of RNA-seq and TMT between HE and LE groups; (E). Share KEGG analysis of RNA-seq and TMT between HE and LE groups.
FIGURE 4
FIGURE 4
Identification and verification of differentially expressed gene and protein. (A). Changes in DEG expression validated by qRT-PCR. Comparison results of the 14 mRNAs using the qRT-PCR and RNA-seq; (B) changes of DEP expression validated by Western Blot. Comparison results of the eight proteins using the Western Blotting and TMT; (C). Western Blot result of part DEPs; (D) relative expression of KRT14 in different tissues; (E) relative expression of ANXA2 in different tissues; (F) relative expression of DKK3 in different tissues.
FIGURE 5
FIGURE 5
Effect of KRT14 on calcium metabolism and deposition of calcium carbonate in eggshell. (A–E): Expression levels of KRT14, OC116, CALB1, BST1, and OC17 mRNA.
See this image and copyright information in PMC

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