Differential Membrane Protein Profile in Bovine X- and Y‑Sperm

The
aim of this study was to understand the molecular mechanisms
behind the biological differences of X- and Y-sperm and to screen
the sex-specific candidate antigen proteins for sexed semen production.
To this end, we investigated differential expression of total membrane
proteins of the two sperm types by using high-purity X- and Y-sperm
from 20 Holstein bulls and applying the label-free proteomic technique;
1521 proteins were identified. In the X-sperm group, 8 and 23 proteins
were significantly up- and down-regulated, respectively. In the X-
and the Y-sperm group, 151 and 88 proteins were specifically expressed,
respectively. These were overexpressed in the dynamic changes of the
actin cytoskeleton, and cell senescence/apoptosis induced by the immune
response, and could result in differences in the state, size, and
immune sensitivity of the X-/Y-sperm membranes. The prediction of
transmembrane structure, subcellular localization, and Western blotting
validation results showed that the CLRN3 and SCAMP1 proteins were
cell surface specific antigens of X- and Y-sperm, respectively. Our
findings help explain the molecular mechanism behind the biological
differences of X-/Y-sperm and lay the foundation for application of
immunological methods to produce sex-sorted semen and control livestock
sex. Data are available via ProteomeXchange with identifier PXD019435.