Key Laboratory of Applied Marine Biotechnology by the Ministry of Education and Key Laboratory of Marine Biotechnology of Zhejiang Province, School of Marine Sciences, Ningbo University, Ningbo 315822, China.
Int J Mol Sci. 2023 Feb 22;24(5):4358. doi: 10.3390/ijms24054358.
is a very important marine economic species, and its aquaculture industry has been developing rapidly. However, the phenomenon of marine wild capture of and germplasm degradation has become increasingly serious. It is necessary to develop the artificial farming industry and carry out germplasm resource protection, for which sperm cryopreservation technology is an effective method. This research compared three methods (mesh-rubbing, trypsin digestion, and mechanical grinding) for acquiring free sperm, and the best method was mesh-rubbing. Then, the optimal cryopreservation conditions were selected, and the optimal formulation was sterile calcium-free artificial seawater, the optimal cryoprotectant was 20% glycerol, and the best equilibrium time was 15 min at 4 °C. The optimal cooling program was suspending the straws at 3.5 cm on the liquid nitrogen surface for 5 min and then storing them in liquid nitrogen. Finally, the sperm were thawed at 42 °C. However, the expression of sperm-related genes and the total enzymatic activities of frozen sperm were significantly decreased ( < 0.05), which showed that sperm cryopreservation damaged the sperm. Our study improves the sperm cryopreservation technology and the yield of aquaculture in Additionally, the study provides a certain technical basis for the establishment of a sperm cryopreservation library of crustaceans.
是一种非常重要的海洋经济物种，其水产养殖业发展迅速。然而，海洋野生捕捞和种质退化的现象日益严重。有必要发展人工养殖产业，开展种质资源保护，精子冷冻保存技术是一种有效的方法。本研究比较了三种获取游离精子的方法（网搓法、胰酶消化法和机械研磨法），最佳方法是网搓法。然后，选择了最佳的冷冻保存条件，最佳配方是无菌无钙人工海水，最佳冷冻保护剂是 20%甘油，最佳平衡时间为 4°C 下 15 分钟。最佳冷却方案是将 straw 在液氮表面 3.5 厘米处悬挂 5 分钟，然后储存在液氮中。最后，将精子在 42°C 下解冻。然而，精子相关基因的表达和冷冻精子的总酶活性显著降低（<0.05），这表明精子冷冻保存会损害精子。我们的研究提高了水产养殖的精子冷冻保存技术和产量。此外，本研究为甲壳类动物精子冷冻保存库的建立提供了一定的技术基础。
