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首页> 外文期刊>Theriogenology >Flow cytometry and ultrastructure of cryopreserved red seabream (Pagrus major) sperm
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Flow cytometry and ultrastructure of cryopreserved red seabream (Pagrus major) sperm

机译：冷冻保存的红鲷（Pagrus major）精子的流式细胞仪和超微结构

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The objectives were to assess motility, fertilizing capacity, structural integrity, and mitochondrial function in fresh versus frozen-thawed (15% DMSO was used as a cryoprotectant) sperm from red seabream (Pagrus major). Mean (+/-S.D.) rates of motility, fertilization and hatching of frozen-thawed sperm were 81.0+/-5.4, 92.8+/-1.9, and 91.8+/-5.2%, respectively; for fresh sperm, they were 87.5+/-7.7, 95.8+/-2.4, and 93.8+/-4.2%. Although motility was lower in frozen-thawed versus fresh sperm (P<0.05), there was no effect (P>0.05) of cryopreservation on fertilization or hatching. Based on scanning and transmission electron microscopy, 77.8+/-5.6% of fresh sperm had normal morphology, whereas for frozen-thawed sperm, 63.0+/-7.2% had normal morphology, 20.6+/-3.1% were slightly damaged (e.g. swelling or rupture of head, mid-piece and tail region as well as mitochondria), and 16.4+/-4.2% were severely damaged. Sperm were stained with propidium iodide and Rhodamine 123 to assess plasma membrane integrity and mitochondrial function, respectively, and examined with flow cytometry. For fresh sperm, 83.9% had an intact membrane and functional mitochondria, whereas for frozen-thawed sperm, 74.8% had an intact membrane and functional mitochondria, 12.7% had a damaged membrane, 9.9% had nonfunctional mitochondria, and 2.6% had both a damaged membrane and nonfunctional mitochondria. In conclusion, ultrastructure and flow cytometry were valuable for assessment of frozen-thawed sperm quality; cryopreservation damaged the sperm but fertilizing ability was not significantly decreased.
机译：目的是评估红鲷鱼（Pagrus major）的新鲜与冷冻融化（15％DMSO作为冷冻保护剂）精子的活力，受精能力，结构完整性和线粒体功能。冻融精子的平均运动度（+/- S.D。）为81.0 +/- 5.4、92.8 +/- 1.9和91.8 +/- 5.2％。新鲜精子的检出率为87.5 +/- 7.7、95.8 +/- 2.4和93.8 +/- 4.2％。尽管冻融后的活力比新鲜精子低（P <0.05），但冷冻保存对受精或孵化没有影响（P> 0.05）。根据扫描和透射电子显微镜，新鲜精子的形态为正常的77.8 +/- 5.6％，而冻融精子的形态为正常的63.0 +/- 7.2％，轻度受损的为20.6 +/- 3.1％（例如肿胀）或头部，中段和尾部区域以及线粒体破裂），严重损坏率为16.4 +/- 4.2％。精子分别用碘化丙锭和若丹明123染色以评估质膜完整性和线粒体功能，并用流式细胞仪检查。新鲜精子的完整膜和功能性线粒体为83.9％，而冻融精子的完整膜和功能性线粒体为74.8％，膜受损的为12.7％，无功能线粒体为9.9％，无功能线粒体为2.6％。受损的膜和无功能的线粒体。总之，超微结构和流式细胞仪对于评估冻融精子质量具有重要价值。冷冻保存会损害精子，但受精能力并没有明显降低。

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