Identification of multiple male reproductive tract-specific proteins that regulate sperm migration through the oviduct in mice (Ikawa Lab in PNAS)

CRISPR/Cas9-mediated genome editing technology enables re- searchers to efficiently generate and analyze genetically modified animals. We have taken advantage of this game-changing tech- nology to uncover essential factors for fertility. In this study, we generated knockouts (KOs) of multiple male reproductive organ- specific genes and performed phenotypic screening of these null mutant mice to attempt to identify proteins essential for male fertility. We focused on making large deletions (dels) within 2 gene clusters encoding cystatin (CST) and prostate and testis expressed (PATE) proteins and individual gene mutations in 2 other gene families encoding glycerophosphodiester phosphodi- esterase domain (GDPD) containing and lymphocyte antigen 6 (Ly6)/Plaur domain (LYPD) containing proteins. These gene fami- lies were chosen because many of the genes demonstrate male reproductive tract-specific expression. Although Gdpd1 and Gdpd4 mutant mice were fertile, disruptions of Cst and Pate gene clusters and Lypd4 resulted in male sterility or severe fertility defects sec- ondary to impaired sperm migration through the oviduct. While absence of the epididymal protein families CST and PATE affect the localization of the sperm membrane protein A disintegrin and metallopeptidase domain 3 (ADAM3), the sperm acrosomal mem- brane protein LYPD4 regulates sperm fertilizing ability via an ADAM3-independent pathway. Thus, use of CRISPR/Cas9 technol- ogies has allowed us to quickly rule in and rule out proteins required for male fertility and expand our list of male-specific proteins that function in sperm migration through the oviduct.

This article will be published in Proc Natl Acad Sci USA (PNAS), on the week of Aug 26, 2019.

Title：“Identification of multiple male reproductive tract-specific proteins that regulate sperm migration through the oviduct in mice”

Authors：Fujihara Y, Noda T, Kobayashi K, Oji A, Kobayashi S, Matsumura T, Larasati T, Oura S, Kojima-Kita K, Yu Z, Matzuk MM, and Ikawa M.