Department of Veterinary Medical Sciences, Alma Mater Studiorum-University of Bologna, Bologna, Italy.
Biotechnology of Animal and Human Reproduction (TechnoSperm), Unit of Cell Biology, Department of Biology, Faculty of Sciences, Institute of Food and Agricultural Technology, University of Girona, Girona, Spain.
Reprod Domest Anim. 2023 Jan;58(1):184-188. doi: 10.1111/rda.14261. Epub 2022 Sep 30.
This study evaluated the bioenergetic map of mitochondria metabolism in cryopreserved bovine sperm. The detected oligomycin-sensitive basal respiration supported ATP production; frozen-thawed spermatozoa were found to have a coupling efficiency higher than 0.80. Cell respiration, however, was not stimulated by the protonophoric action of FCCP, as its titration with 1, 2, 4 and 6 μM did not stimulate the uncoupling activity on oxidative phosphorylation as highlighted by unresponsive oxygen consumption. The unusual effect on the stimulation of maximal respiration was not related to fibronectin- or PDL-coated plates used for cellular metabolism analysis. Conversely, irradiation of frozen-thawed bovine sperm with the red light improved mitochondrial parameters. In effect, the maximal respiration of red-light-stimulated sperm in PDL-coated plates was higher than the non-irradiated. In spite of this, red-light irradiation had no impact on membrane integrity and mitochondrial activity evaluated by epifluorescence microscopy.
本研究评估了冷冻牛精子中线粒体代谢的生物能量图谱。检测到的寡霉素敏感基础呼吸支持 ATP 产生；冷冻 - 解冻的精子具有高于 0.80 的偶联效率。然而，质子载体 FCCP 的作用并没有刺激细胞呼吸，因为用 1、2、4 和 6 μM 滴定并没有刺激氧化磷酸化的解偶联活性，如耗氧量无反应所强调的那样。对最大呼吸刺激的异常影响与用于细胞代谢分析的纤连蛋白或 PDL 涂层板无关。相反，用红光照射冷冻解冻的牛精子可改善线粒体参数。实际上，PDL 涂层板中经红光刺激的精子的最大呼吸高于未照射的精子。尽管如此，红光照射对通过荧光显微镜评估的膜完整性和线粒体活性没有影响。
