Abstract
BackgroundIn previous study, we performed next-gene sequencing to investigate the differentially expressed transcripts of bovine follicular granulosa cells (GCs) at dominant follicle (DF) and subordinate follicle (SF) stages during first follicular wave. Present study is designed to further identify the key regulatory proteins and signaling pathways associated with follicular development using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) and multi-omics data analysis approach.MethodsDF and SF from three cattle were collected by daily ultrasonography. The GCs were isolated from each follicle, total proteins were digested by trypsin, and then proteomic analyzed via LC-MS/MS, respectively. Proteins identified were retrieved from Uniprot-COW fasta database, and differentially expressed proteins were used to functional enrichment and KEGG pathway analysis. Proteome data and transcriptome data obtained from previous studies were integrated.ResultsTotal 3409 proteins were identified from 30,321 peptides (FDR ≤0.01) obtained from LC-MS/MS analysis and 259 of them were found to be differentially expressed at different stage of follicular development (fold Change > 2, P < 0.05). KEGG pathway analysis of proteome data revealed important signaling pathways associated with follicular development, multi-omics data analysis results showed 13 proteins were identified as being differentially expressed in DF versus SF.ConclusionsThis study represents the first investigation of transcriptome and proteome of bovine follicles and offers essential information for future investigation of DF and SF in cattle. It also will enrich the theory of animal follicular development.
Highlights
Mono-ovulatory species such as cattle undergoes 2–3 follicular wave during one estrous/menstrual cycle
Several studies have been performed in the past to understand the physiological regulation of follicular development modulation and ovulation mechanism, such as gonadotrophin interacts with follicular growth factor [4,5,6], the relationship between apoptosis of follicle granulosa cells (GCs) and follicle atresia [7,8,9,10], the influence of hormone such as estrogen and progesterone [11], the role of growth factor such as vascular endothelial growth factor (VEGF) and insulin-like growth factors 1 (IGF-1) [12,13,14], the effect of concentration of reactive oxygen [15]
This is the first study where we characterized the proteome of bovine GCs at dominant follicle (DF) and subordinate follicle (SF) in first follicular wave and performed an integrated analysis of protein abundance and transcript level to reveal the molecular regulation of bovine follicular development
Summary
Mono-ovulatory species such as cattle undergoes 2–3 follicular wave during one estrous/menstrual cycle. Transcriptome of follicles at different developmental stage have been analyzed to understand the molecular mechanism regulating bovine follicular development [16]. Characterizing the functionally available proteins resulting from mRNA translation would provide novel insight into the signaling pathways and biomarkers of follicular developmental process in cattle. This is the first study where we characterized the proteome of bovine GCs at DF and SF in first follicular wave and performed an integrated analysis of protein abundance and transcript level to reveal the molecular regulation of bovine follicular development. Present study is designed to further identify the key regulatory proteins and signaling pathways associated with follicular development using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) and multi-omics data analysis approach
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