ABSTRACT: LC-MS/MS of Prss37 WT and KO mice male reproductive tract.
Title: Molecular dissection and testing of PRSS37 function through LC-MS/MS and the generation of a PRSS37 humanized mouse model
Authors: Courtney Sutton1,2, Kaori Nozawa1,2, Katarzyna Kent1,2, Alexander Saltzman3, Mei Leng3, Sureshbabu Nagarajan1,2, Anna Malovannaya3,4, Masahito Ikawa5,6, Thomas X. Garcia1,2,7, and Martin M. Matzuk1,2,*
Email Addresses and ORCID IDs:
Courtney Sutton: courtney.sutton@bcm.edu, 0000-0001-9994-990X
Kaori Nozawa: nozawa.kaori.029@gmail.com, 0000-0002-2256-278X
Katarzyna Kent: kkent@bcm.edu, 0000-0002-4528-3425
Alexander Saltzman: alexander.saltzman@bcm.edu, 0000-0003-4166-9555
Mei Leng: mleng@bcm.edu, 0000-0001-7231-8665
Sureshbabu Nagarajan: nsureshh@gmail.com, 0000-0003-3602-3709
Anna Malovannaya: malovann@bcm.edu, 0000-0003-2953-6485
Masahito Ikawa: ikawa@biken.osaka-u.ac.jp, 0000-0001-9859-6217
Thomas X. Garcia: thomas.garcia@bcm.edu, 0000-0001-7761-2660
Martin M. Matzuk: mmatzuk@bcm.edu, 0000-0002-1445-8632
*Correspondence to: Martin M. Matzuk, M.D., Ph.D.
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.
Martin M. Matzuk 