Asturiano J F, Marco-Jiménez F, Pérez L, Balasch S, Garzón D L, Peñaranda D S, Vicente J S, Viudes-de-Castro M P, Jover M
Grupo de Investigación en Recursos Acuícolas, Departamento de Ciencia Animal, Universidad Politécnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain.
Theriogenology. 2006 Sep 1;66(4):1012-20. doi: 10.1016/j.theriogenology.2006.02.041. Epub 2006 Apr 3.
Fish sperm quality has traditionally been estimated by subjective evaluation of motility and sperm concentration. Alternative methods for evaluation of sperm quality have been developed in the last decade and enable estimation of spermatozoa head morphometry, membrane integrity and mitochondrial function. Weekly injections of human chorionic gonadotropin (hCG) induced spermiation in farmed male European eels. The milt volume increased from the 5th to 12th weeks. Sperm concentration significantly increased from the 5th week, reaching the highest values at the 8th week, while best motility results were registered at the 9th week of treatment. Coinciding with these intervals, the percentage of dead spermatozoa determined with Hoechst staining showed a reduction in the 8th to 11th weeks of treatment, while the percentage of mitochondrial functionality determined by JC-1 staining did not show a similar pattern. The automatic sperm morphology analysis (ASMA) of the spermatozoa head length, width, area and perimeter showed a significant growth from the 5th to 8th weeks. However, the analysis of isolated descriptive parameters may be difficult to understand because there is a variability in these parameters for each week, making knowledge of the growth kinetic complex. The global size of the spermatozoa head was calculated by applying principal component analysis (PCA), because this method establishes new components that make the interpretation of results easier, allowing a whole interpretation of the changes in the cell morphology. PC1 defines the global head size and shows a significant increase between the 5th and 8th weeks of treatment, showing shorter changes until 12th week. PC2 shows a significant increase in the spermatozoa width from the 5th to 7th weeks. Considering the results of the variations in the principal components defining European eel spermatozoa morphometry, it may be concluded that hCG maturative treatment produced thick cells during the first weeks of spermiation, and subsequent samplings showed an increase in cell width and length. These changes in sperm morphometry coincide with the highest sperm quality assessed as sperm motility and concentration, as well as with the best results obtained in previous studies reporting the best sperm quality between weeks 8 and 10 of hCG treatment. These results support the use of ASMA and Hoechst staining techniques as alternative methods for the evaluation of fish sperm quality.
传统上，鱼类精子质量是通过对活力和精子浓度的主观评估来估计的。在过去十年中，已经开发出了评估精子质量的替代方法，这些方法能够估计精子头部形态、膜完整性和线粒体功能。每周注射人绒毛膜促性腺激素（hCG）可诱导养殖的雄性欧洲鳗鱼排精。精液体积从第5周增加到第12周。精子浓度从第5周开始显著增加，在第8周达到最高值，而最佳活力结果出现在治疗的第9周。与这些时间段一致，用Hoechst染色法测定的死精子百分比在治疗的第8至11周有所下降，而用JC-1染色法测定的线粒体功能百分比则没有呈现类似的模式。对精子头部长度、宽度、面积和周长进行的自动精子形态分析（ASMA）显示，从第5周到第8周有显著增长。然而，对孤立的描述性参数进行分析可能难以理解，因为这些参数在每周都存在变异性，使得对生长动力学的了解变得复杂。通过应用主成分分析（PCA）来计算精子头部的整体大小，因为这种方法建立了新的成分，使结果的解释更容易，从而能够对细胞形态的变化进行整体解释。主成分1（PC1）定义了整体头部大小，在治疗的第5周和第8周之间显示出显著增加，直到第12周变化较小。主成分2（PC2）显示精子宽度从第5周到第7周有显著增加。考虑到定义欧洲鳗鱼精子形态的主成分变化结果，可以得出结论，hCG成熟处理在排精的最初几周产生了较粗的细胞，随后的采样显示细胞宽度和长度增加。精子形态的这些变化与作为精子活力和浓度评估的最高精子质量相吻合，也与先前研究中报道的hCG治疗第8至10周精子质量最佳的最佳结果相吻合。这些结果支持使用ASMA和Hoechst染色技术作为评估鱼类精子质量的替代方法。
