PhospholipaseA2VI mediates the lipid peroxidation-ferroptosis axis to regulate rooster sperm motility.

We identified individuals with substantial differences in sperm motility among high-motility sperm samples preserved under various conditions. Phospholipase A2 group VI (PLA2G6) was upregulated in the low-motility group (LMG) by integrating seminal plasma metabolome and epididymal transcriptome data. LMG exhibited increased malondialdehyde and Fe²⁺ levels, whereas the sperm's membrane integrity and glutathione peroxidase 4's activity were reduced. Targeted PLA2G6 activity inhibition led to a substantial reduction in PLA2G6 activity and lysophosphatidylcholine levels in the SP, accompanied by improved sperm motility and membrane integrity, extended in vitro survival, and increased fertilization efficiency. PLA2G6 promoted sperm motility loss by hydrolyzing phosphatidylcholine to generate lysophosphatidylcholine and polyunsaturated fatty acids, ultimately resulting in lipid peroxidation and ferroptosis. We provide new insights into the mechanisms underlying sperm motility regulation, a potential strategy for extending the artificial insemination window, and developing semen preservation technologies based on ferroptosis regulation.