ORIGINAL RESEARCH article
Front. Cell Dev. Biol.
Sec. Molecular and Cellular Reproduction
This article is part of the Research TopicUnlocking Fertility: Advances in Fertilization Research and Innovations in Assisted Reproductive TechnologiesView all 4 articles
Exploring the effect of skim milk on the membrane stability of frozen-thawed Inner Mongolia cashmere goat sperm based on proteomics
Provisionally accepted
- Inner Mongolia Agricultural University, Hohhot, China
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Inner Mongolia cashmere goat, a unique dual-purpose breed in China for both cashmere and meat, experiences a sharp decline in semen motility after cryopreservation, severely constraining genetic improvement and germplasm propagation. In this study, skim milk was added stepwise (2–3.6 %) to an egg yolk–soy lecithin basal extender, with 2.8 % identified as the optimal concentration; TMT quantitative proteomics coupled with PRM targeted validation were employed to elucidate the homeostatic mechanisms of post-thaw sperm membranes. Results showed that 2.8 % skim milk increased post-thaw sperm motility to 68.23 %, reduced ultrastructural abnormalities, elevated acrosomal integrity by 18.7 %, and decreased lipid peroxidation by 29 % (P < 0.05). Proteomic profiling identified 32 differentially expressed proteins (DEPs); GO enrichment analysis revealed significant enrichment in processes related to purine ribonucleoside triphosphate (NTP) metabolism and transmembrane transporter activity. KEGG pathway analysis indicated predominant enrichment in energy metabolism and signal transduction pathways. PRM confirmed that NDUFA8, PGAM2, ACTL7A, PRXL2B, ATP6V0C, and LELP1 exhibited expression patterns consistent with the proteomics data, serving as core biomarkers for skim milk-mediated stabilization of sperm membranes. This study provides the first proteomic-level evidence that skim milk enhances cryotolerance of Inner Mongolia cashmere goat spermatozoa by modulating an energy-membrane protein network, establishing molecular biomarkers for optimizing semen cryopreservation protocols.
Keywords: Inner Mongolia Cashmere Goat, Semen Cryopreservation, skim milk, Cryodamage, Proteomics
Received: 09 Sep 2025; Accepted: 08 Dec 2025.
Copyright: © 2025 Xue, Xu, Li, Zhang and Su. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Shan-hui Xue
Rui Su
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