MALDI-IMS combined with shotgun proteomics identify and localize new factors in male infertility.
Shibojyoti LahiriWasim AftabLena WalentaLeena StraussMatti PoutanenMayerhofer ArturAxel ImhofPublished in: Life science alliance (2021)
Spermatogenesis is a complex multi-step process involving intricate interactions between different cell types in the male testis. Disruption of these interactions results in infertility. Combination of shotgun tissue proteomics with MALDI imaging mass spectrometry is markedly potent in revealing topological maps of molecular processes within tissues. Here, we use a combinatorial approach on a characterized mouse model of hormone induced male infertility to uncover misregulated pathways. Comparative testicular proteome of wild-type and mice overexpressing human P450 aromatase (AROM+) with pathologically increased estrogen levels unravels gross dysregulation of spermatogenesis and emergence of pro-inflammatory pathways in AROM+ testis. In situ MS allowed us to localize misregulated proteins/peptides to defined regions within the testis. Results suggest that infertility is associated with substantial loss of proteomic heterogeneity, which define distinct stages of seminiferous tubuli in healthy animals. Importantly, considerable loss of mitochondrial factors, proteins associated with late stages of spermatogenesis and steroidogenic factors characterize AROM+ mice. Thus, the novel proteomic approach pinpoints in unprecedented ways the disruption of normal processes in testis and provides a signature for male infertility.
Keyphrases
- mass spectrometry
- wild type
- liquid chromatography
- polycystic ovary syndrome
- high resolution
- germ cell
- gas chromatography
- mouse model
- high performance liquid chromatography
- capillary electrophoresis
- single cell
- endothelial cells
- label free
- high fat diet induced
- gene expression
- high glucose
- stem cells
- oxidative stress
- adipose tissue
- insulin resistance
- cell therapy
- ms ms
- drug induced
- pluripotent stem cells
- single molecule
- stress induced