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具有高精子 DNA 碎片化指数的种公牛的精子表型特征和输卵管结合能力发生改变。

Sperm phenotypic characteristics and oviduct binding ability are altered in breeding bulls with high sperm DNA fragmentation index.

机构信息

Theriogenology Laboratory, Southern Regional Station of ICAR-National Dairy Research Institute, Bengaluru, 560030, Karnataka, India.

Theriogenology Laboratory, Southern Regional Station of ICAR-National Dairy Research Institute, Bengaluru, 560030, Karnataka, India.

出版信息

Theriogenology. 2021 Sep 15;172:80-87. doi: 10.1016/j.theriogenology.2021.06.006. Epub 2021 Jun 9.

Abstract

In the present study, we standardized an in vitro oviduct explants model for cattle and assessed the oviduct explants binding ability and phenotypic characteristics of spermatozoa obtained from breeding bulls with high- and low-sperm DNA fragmentation index (DFI%). Cryopreserved spermatozoa from Holstein Friesian crossbred breeding bulls (n = 45) with known field fertility were assessed for DFI% and were classified into either high DFI% or low DFI% category. Flow cytometry was used to assess sperm membrane integrity, acrosome reaction status, mitochondrial membrane potential and intracellular calcium concentrations. It was found that spermatozoa from bulls with low DFI% had significantly higher (P < 0.05) membrane integrity, acrosome intactness, and mitochondrial membrane potential. To assess the sperm oviduct binding ability, oviduct explants were prepared by incubating the oviduct cells overnight in TCM-199 medium at 38.5 °C under 5% CO. Different sperm concentrations and times of incubation were evaluated and found that 2 million spermatozoa and 1-h incubation yielded high binding index (BI). The BI was also significantly (P < 0.01) higher (>2 times) in the bulls with low-DFI% as compared to high DFI% bulls. The correlation between binding index and DFI% was negative and significant (r = -0.528; P < 0.05). Further, the binding index was positively correlated with conception rate (r = 0.703), intact sperm membrane (r = 0.631) and mitochondrial membrane potential (r = 0.609). It is inferred that sperm phenotypic characteristics and oviduct binding ability are impaired in breeding bulls with high sperm DFI%, which might be associated with low conception rates in these bulls.

摘要

在本研究中,我们建立了牛的体外输卵管组织块模型,并评估了来自精子 DNA 碎片化指数(DFI%)高和低的种公牛的输卵管组织块结合能力和精子的表型特征。我们对具有已知田间生育力的荷斯坦弗里生杂交种公牛的冷冻精子进行了 DFI%评估,并将其分为高 DFI%或低 DFI%类别。使用流式细胞术评估了精子膜完整性、顶体反应状态、线粒体膜电位和细胞内钙离子浓度。结果发现,DFI%低的公牛的精子具有更高的(P<0.05)膜完整性、顶体完整率和线粒体膜电位。为了评估精子与输卵管的结合能力,我们通过将输卵管细胞在 38.5°C、5% CO2 下在 TCM-199 培养基中孵育过夜来制备输卵管组织块。评估了不同的精子浓度和孵育时间,发现 200 万个精子和 1 小时孵育产生高的结合指数(BI)。与 DFI%高的公牛相比,DFI%低的公牛的 BI 也显著更高(P<0.01)(>2 倍)。结合指数与 DFI%之间呈负相关且具有统计学意义(r=-0.528;P<0.05)。此外,结合指数与妊娠率呈正相关(r=0.703),与完整的精子膜呈正相关(r=0.631),与线粒体膜电位呈正相关(r=0.609)。可以推断,DFI%高的种公牛的精子表型特征和与输卵管的结合能力受损,这可能与这些公牛的低妊娠率有关。

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