4D label-free quantitative proteomic analysis identifies CRABP1 as a novel candidate gene for litter size in rabbits

Zhiyuan Bao; Yang Chen; Jiali Li; Jiawei Cai; Jie Yang; Pin Zhai; Bohao Zhao; Xinsheng Wu

doi:10.1093/biolre/ioae038

4D label-free quantitative proteomic analysis identifies CRABP1 as a novel candidate gene for litter size in rabbits

Biol Reprod. 2024 Mar 13:ioae038. doi: 10.1093/biolre/ioae038. Online ahead of print.

Authors

Zhiyuan Bao¹, Yang Chen^{1

2}, Jiali Li¹, Jiawei Cai¹, Jie Yang¹, Pin Zhai³, Bohao Zhao^{1

2}, Xinsheng Wu^{1

2}

Affiliations

¹ College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, Jiangsu, People's Republic of China.
² Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, 225009 Yangzhou, Jiangsu, China.
³ Institute of Animal Science, Jiangsu Academy of Agricultural Sciences.

PMID: 38478424
DOI: 10.1093/biolre/ioae038

Abstract

In commercial rabbit breeding, litter size is a crucial reproductive trait. This trait directly determines the reproductive ability of female rabbits and is crucial for evaluating the production efficiency. We here compared differentially expressed proteins of in the ovary tissue from New Zealand female rabbits with high (H) and low (L) litter sizes by using 4D label-free quantitative proteomic technology and identified 92 differential proteins. The biological functions of these proteins were revealed through gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Most distributions of GO and KEGG were related to reproduction, growth development, and metabolism. Furthermore, a novel candidate gene Cellular Retinoic Acid Binding Protein-1 (CRABP1), which was highly expressed in the L group, was selected for further biological function verification. The Cell Counting Kit-8 assay and flow cytometry analysis revealed that CRABP1 can promote granulosa cell (GC) apoptosis and inhibit GC proliferation. Furthermore, qRT-PCR and western blotting analysis revealed that CRABP1 regulates the genes (HSD17B1, Wnt-10b, FSHR, TAF4B, BMP15, and BMP6) and protein (Wnt-10b) associated with steroid hormone synthesis and follicle development. The PCR product direct sequencing method revealed single nucleotide polymorphisms in the core promoter region of CRABP1. Luciferase activity assays revealed that the transcriptional activity of the GG genotype was significantly higher than that of the TT or TG genotype. Different genotypes are accompanied by changes in transcription factors, which indicates that T-359G polymorphism can regulate CRABP1 expression. In general, we identified litter size-related genes and revealed the mechanism underlying the effect of CRABP1 on litter size. CRABP1 serves as a key factor in the reproductive capacity of rabbits and can act as a molecular biomarker for the breeding of New Zealand rabbits.

Keywords: CRABP1; Litter size; Ovary; Proteome; Rabbits.