Molecular Dissection and Testing of PRSS37 Function Through Lc-Ms/Ms and the Generation of a PRSS37 Humanized Mouse Model

Authors

Courtney Sutton
Kaori Nozawa
Katarzyna Kent
Alexander Saltzman
Mei Leng
Sureshbabu Nagarajan
Anna Malovannaya
Masahito Ikawa
Thomas X Garcia
Martin M Matzuk

Publication Date

7-14-2023

Journal

Scientific Reports

DOI

10.1038/s41598-023-37700-1

PMID

37452050

PMCID

PMC10349139

PubMedCentral® Posted Date

7-14-2023

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Male, Humans, Mice, Animals, Peptide Hydrolases, Chromatography, Liquid, Proteomics, Tandem Mass Spectrometry, Semen, Mice, Transgenic, Mice, Knockout, Endopeptidases, Infertility, Male, Urogenital models, Experimental organisms, Genetic engineering, Mass spectrometry, Proteomic analysis

Abstract

The quest for a non-hormonal male contraceptive pill for men still exists. Serine protease 37 (PRSS37) is a sperm-specific protein that when ablated in mice renders them sterile. In this study we sought to examine the molecular sequelae of PRSS37 loss to better understand its molecular function, and to determine whether human PRSS37 could rescue the sterility phenotype of knockout (KO) mice, allowing for a more appropriate model for drug molecule testing. To this end, we used CRISPR-EZ to create mice lacking the entire coding region of Prss37, used pronuclear injection to create transgenic mice expressing human PRSS37, intercrossed these lines to generate humanized mice, and performed LC-MS/MS of KO and control tissues to identify proteomic perturbances that could attribute a molecular function to PRSS37. We found that our newly generated Prss37 KO mouse line is sterile, our human transgene rescues the sterility phenotype of KO mice, and our proteomics data not only yields novel insight into the proteome as it evolves along the male reproductive tract, but also demonstrates the proteins significantly influenced by PRSS37 loss. In summary, we report vast biological insight including insight into PRSS37 function and the generation of a novel tool for contraceptive evaluation.

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