Qin Xinxi, Zhang Haisen, Liu Tian, Cui Zhenliang, Gao Kangkang, Lin Pengfei, Jin Yaping
Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
Key Laboratory of Animal Biotechnology of the Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
Animals (Basel). 2025 May 15;15(10):1430. doi: 10.3390/ani15101430.
Follicle stimulating hormone (FSH) stands as one of the most prevalently used reproductive hormones in the field of animal-assisted reproduction. Conventionally, pituitary FSH is sourced from the heterologous pituitary glands of pigs and sheep procured from slaughterhouses, and it typically exists in the form of crude FSH. The specific challenges inherent in FSH-based assisted reproduction drugs has significantly spurred the interest in exploring novel alternatives, aiming to reduce the reliance on these traditional sources in relevant production processes. In this study, the α- and β-FSH genes were retrieved from pituitary cDNA libraries. These genes were selected to construct a recombinant protein-the novel cow recombinant FSH (CrFSH)-through the application of the homologous recombination method. Notably, the β-subunit was extended by a carboxy-terminal peptide (CTP). After successfully integrating the two genes into Chinese hamster ovary (CHO) cells, the recombinant protein (approximately 33 kDa) in the culture supernatant was detected using Western blotting (WB). The results of the GCs proliferation experiment indicated that both 1.2 µg/mL pFSH and 20-20,000 ng/mL CrFSH could significantly promote the proliferation of GCs in vitro. Remarkably, on the 4th day after treatment, 20 ng/mL of CrFSH had a higher GCs proliferation rate than 1.2 μg/mL of pFSH ( < 0.001). Additionally, cyclic adenosine monophosphate (cAMP) induction assay in GCs unequivocally confirmed that CrFSH possesses superior activity compared to pFSH. These findings underscore that this recombinant protein holds great potential as a promising candidate for FSH production in assisted reproduction approaches for dairy herds.
促卵泡激素（FSH）是动物辅助生殖领域中使用最为普遍的生殖激素之一。传统上，垂体FSH来源于从屠宰场采购的猪和羊的异种垂体，并且通常以粗制FSH的形式存在。基于FSH的辅助生殖药物所固有的特定挑战显著激发了人们探索新型替代物的兴趣，旨在减少相关生产过程中对这些传统来源的依赖。在本研究中，从垂体cDNA文库中检索到α-和β-FSH基因。通过应用同源重组方法选择这些基因来构建一种重组蛋白——新型牛重组FSH（CrFSH）。值得注意的是，β亚基通过羧基末端肽（CTP）进行了延伸。在将这两个基因成功整合到中国仓鼠卵巢（CHO）细胞中后，使用蛋白质免疫印迹法（WB）检测培养上清液中的重组蛋白（约33 kDa）。颗粒细胞（GCs）增殖实验结果表明，1.2 μg/mL的垂体FSH（pFSH）和20 - 20,000 ng/mL的CrFSH均可显著促进体外GCs的增殖。值得注意的是，在处理后的第4天，20 ng/mL的CrFSH的GCs增殖率高于1.2 μg/mL的pFSH（<0.001）。此外，在GCs中进行的环磷酸腺苷（cAMP）诱导试验明确证实，CrFSH比pFSH具有更高的活性。这些发现强调，这种重组蛋白作为奶牛群辅助生殖方法中FSH生产的有前景的候选物具有巨大潜力。
