2. Academic Validation
  3. Autophagic Degradation of GPX4 Mediates Ferroptosis During Sheep Sperm Cryopreservation

Autophagic Degradation of GPX4 Mediates Ferroptosis During Sheep Sperm Cryopreservation

  • Vet Sci. 2025 May 19;12(5):490. doi: 10.3390/vetsci12050490.
Boyuan Li 1,  Erhan Hai 1,  Yukun Song 1,  Jiaxin Zhang 1
Affiliations

Affiliation

  • 1 Inner Mongolia Key Laboratory of Sheep & Goat Genetics Breeding and Reproduction, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
PMID: 40431583 DOI: 10.3390/vetsci12050490
Abstract

Ferroptosis is implicated in cryodamage to sheep sperm, potentially due to Glutathione Peroxidase 4 (GPX4) degradation during freezing; however, the pathway underlying GPX4 degradation remains unclear. In this study, a comparison of cryoprotective effects between the Autophagy inhibitor chloroquine (CQ) and the ubiquitination inhibitor MG132 revealed that 5 μM CQ treatment significantly enhanced the motility (p < 0.01) and sperm plasma membrane integrity rate (p < 0.01) of frozen-thawed sperm; no protective effects were observed in any MG132 treatment group. Mechanistic analysis indicated that CQ treatment substantially restored GPX4 protein expression (p < 0.01), and concurrently reduced lipid peroxidation (p < 0.01) and free iron ion accumulation (p < 0.01), in frozen-thawed sperm. These findings suggest that GPX4 degradation during cryopreservation occurs via the Autophagy pathway. This study established a ferroptosis-GPX4-autophagy axis during sheep sperm cryopreservation and identified autophagy-mediated GPX4 loss as a potential target for enhancing sperm cryoprotection.

Keywords

GPX4; autophagy; ferroptosis; sperm cryopreservation.

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