Garzón D L, Peñaranda D S, Pérez L, Marco-Jiménez F, Espert X, Müller T, Jover M, Asturiano J F
Departamento de Ciencia Animal, Grupo de Investigación en Recursos Acuícolas, Universidad Politécnica de Valencia, Camino de Vera s/n, Valencia, Spain.
Reprod Domest Anim. 2008 Feb;43(1):99-105. doi: 10.1111/j.1439-0531.2007.00861.x.
The main objective of the present study was to evaluate the influence of pH and bicarbonate concentration in the activation or inhibition of European eel (Anguilla anguilla) spermatozoa and to evaluate the effect of different cryoprotectants: dimethyl sulphoxide (DMSO), acetamide, ethylene glycol, propanol, glycerol and methanol (MeOH). The effect of these factors was evaluated comparing the percentage of motile cells, the percentage of alive cells (by Hoechst staining) and the spermatozoa morphometry pre- and post-cryopreservation (by computer-assisted morphology analysis). Based on the above findings, three cryoprotectants (DMSO, MeOH and glycerol) were chosen and evaluated in two media (P1 and P1 modified) with different concentrations of NaHCO(3) and in the presence or absence of foetal bovine serum (FBS). The effect of these factors was evaluated comparing the percentage of alive and motile cells post-cryopreservation. DMSO was the cryoprotectant showing better results in relation to the percentage of spermatic alive cells post-freezing and caused a smaller modification of the head spermatozoa morphology. The combination of P1-modified medium with DMSO and containing FBS increased slightly but significantly the percentage of motile spermatozoa post-cryopreservation.
本研究的主要目的是评估pH值和碳酸氢盐浓度对欧洲鳗鲡（Anguilla anguilla）精子激活或抑制的影响，并评估不同冷冻保护剂的效果：二甲基亚砜（DMSO）、乙酰胺、乙二醇、丙醇、甘油和甲醇（MeOH）。通过比较活动细胞百分比、活细胞百分比（通过Hoechst染色）以及冷冻保存前后精子形态学（通过计算机辅助形态分析）来评估这些因素的影响。基于上述发现，选择了三种冷冻保护剂（DMSO、MeOH和甘油），并在两种含有不同浓度NaHCO₃的培养基（P1和改良P1）中，在有或无胎牛血清（FBS）的情况下进行评估。通过比较冷冻保存后活细胞和活动细胞的百分比来评估这些因素的影响。DMSO是冷冻保护剂，在冷冻后精子活细胞百分比方面显示出更好的结果，并且对精子头部形态的改变较小。改良P1培养基与DMSO并含有FBS的组合在冷冻保存后活动精子的百分比略有但显著增加。
