Suquet Marc, Gourtay Clémence, Donval Anne, Le Goïc Nelly, Quere Claudie, Malo Florent, Le Grand Jaqueline, Ratiskol Dominique, Mingant Christian, Fauvel Christian
Ifremer, UMR 6539, PFOM Department, Station Expérimentale d'Argenton, Argenton, France.
Ifremer, UMR 6539, PFOM Department, Station Expérimentale d'Argenton, Argenton, France.
Gen Comp Endocrinol. 2016 Apr 1;229:127-31. doi: 10.1016/j.ygcen.2016.02.023. Epub 2016 Mar 2.
Most publications devoted to the cryopreservation of mollusc sperm have focused on the definition of technical protocols, avoiding the description of sperm quality after thawing. The present study investigated the effects of cryopreservation on sperm quality in the great scallop. Wild scallop were fished during the natural spawning period and conditioned in the hatchery before use. Sperm samples were obtained after intragonadal injection of serotonin and cryopreserved using a previously published protocol. Sperm quality was assessed using a panel of four parameters: sperm motility characteristics, using a computer assisted sperm analysis plugin with Image J, intracellular ATP content using an ATP-Lite kit, sperm integrity, using flow cytometry and sperm morphology, using transmission electron microscopy. For each parameter, fresh (control) and thawed spermatozoa were compared. A significant decrease of both the percentage of motile spermatozoa (reduction: 75%) and sperm swimming speed (86%) were observed for thawed sperm compared with fresh sperm. The percentage of living spermatozoa, as assessed using flow cytometry, was significantly lower for thawed sperm (72.4±2.5%) compared with fresh sperm (86.4±1.1). However, no significant difference of intracellular sperm ATP content was observed between fresh and thawed sperm. Post thawing, while some spermatozoa showed little or no morphological differences compared with fresh sperm, others had undergone drastic changes, including swelling of the plasma membrane, structural alterations of the chromatin and damage to mitochondria. In conclusion, the descriptive parameters studied in the present work showed that the quality of thawed great scallop sperm was lower than that of fresh cells but was still sufficient for use in aquaculture programs and sperm cryobanking for this species.
大多数关于软体动物精子冷冻保存的出版物都集中在技术方案的定义上，而避免对解冻后精子质量进行描述。本研究调查了冷冻保存对大扇贝精子质量的影响。在自然产卵期捕捞野生扇贝，并在孵化场进行预处理后使用。通过性腺内注射血清素获得精子样本，并使用先前发表的方案进行冷冻保存。使用一组四个参数评估精子质量：精子活力特征，使用带有Image J的计算机辅助精子分析插件；细胞内ATP含量，使用ATP-Lite试剂盒；精子完整性，使用流式细胞术；精子形态，使用透射电子显微镜。对于每个参数，比较新鲜（对照）精子和解冻精子。与新鲜精子相比，解冻精子的活动精子百分比（降低：75%）和精子游动速度（86%）均显著降低。使用流式细胞术评估，解冻精子的活精子百分比（72.4±2.5%）显著低于新鲜精子（86.4±1.1）。然而，新鲜精子和解冻精子之间未观察到细胞内精子ATP含量的显著差异。解冻后，虽然一些精子与新鲜精子相比形态差异很小或没有差异，但其他精子发生了剧烈变化，包括质膜肿胀、染色质结构改变和线粒体损伤。总之，本研究中所研究的描述性参数表明，解冻后的大扇贝精子质量低于新鲜细胞，但仍足以用于水产养殖计划和该物种的精子冷冻保存。
