Proteomic quantification of native and ECM-enriched mouse ovaries reveals an age-dependent fibro-inflammatory signature

ABSTRACT: The ovarian microenvironment becomes fibrotic and stiff with age due in part to increased stromal collagen and decreased hyaluronan. However, the extracellular matrix (ECM) is a complex network of hundreds of proteins, glycoproteins and glycans which are highly tissue specific and undergo pronounced changes with age. We used a label-free quantitative proteomic methodology to obtain an unbiased and comprehensive protein profile of age-associated alterations of the mammalian ovarian proteome. We analyzed ovaries from reproductively young and old mice using both native and ECM-enriched tissues to specifically interrogate the extracellular matrix. We optimized and validated conditions to enrich for the ECM prior to proteomic analysis and determined that treatment of ovaries with 0.1% SDS for 12.5 hours efficiently removed nuclear material, while maintaining the integrity of the ECM. Following the analysis by data-independent acquisition (DIA), both native and ECM-enriched samples clustered separately based on age, indicating that ovaries from reproductively young and old mice feature clearly distinct proteomic signatures. We identified a total of 4,721 proteins and 383 proteins that are significantly-altered with advanced reproductive age, including key ECM proteins. In addition to ECM proteins, immunoglobulins, proteins that regulate metal homeostasis, and tumor suppressors were highly upregulated with age, whereas downregulated proteins were consistent with the age-dependent decline in fertility. Pathways regulating genomic instability and proteostasis were downregulated with age. Biological processes associated with immune function and ECM remodeling were upregulated with age. These findings provide evidence from a proteomic perspective that the aging ovary provides a fibroinflammatory milieu, and our study suggests target proteins which may drive these age-associated phenotypes for future investigation.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Birgit Schilling  

PROVIDER: MSV000091790 | MassIVE | Mon Apr 24 14:57:00 BST 2023

SECONDARY ACCESSION(S): PXD041789

REPOSITORIES: MassIVE