Morphology and Chromatin Integrity of Stallion Spermatozoa Prepared by Density Gradient and Single Layer Centrifugation Through Silica Colloids
Morphology and Chromatin Integrity of Stallion Spermatozoa Prepared by Density Gradient and Single Layer Centrifugation Through Silica Colloids
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DOI：
10.1111/j.1439-0531.2008.01265.x
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发表时间：
2009-06-01
影响因子：
1.7
通讯作者：
Rodriguez-Martinez, H.
中科院分区：
文献类型：
--
作者：
Morrell, J. M.;Johannisson, A.;Rodriguez-Martinez, H.
目的是研究密度梯度离心（DGC）或单层离心（SLC）是否有可能提高种马精液的质量，包括精子形态和染色质完整性，这两者都与生育能力有关。用10匹马38次射精对两种胶体离心方法进行了评价。比较了未离心样品和离心精子制剂中精子形态、主观能动性和精子染色质完整性。形态正常的精子比例在不同种马之间存在差异（p < 0.001），两种胶体离心方法均提高了正常精子比例（离心前中值67.5%，SLC后中值78%，DGC后中值77%，p < 0.001）。某些异常的发生率降低，如近端胞浆液滴从12.9%降低到8.8% (p < 0.001)，中间片缺陷从5.3%降低到1.4% （p < 0.05）。同样，精子活力和染色质完整性显著改善（p < 0.001），两种离心方法之间无差异。胶体离心可使形态正常、染色质结构正常的种马精子在精子制剂中所占比例增加。新方法SLC在选择形态正常、染色质完整的活动种马精子方面与DGC一样有效。SLC比DGC使用更简单，适合种马人员常规使用，以提高不同授精剂量的种马精子质量。
The objective was to investigate whether it is possible to improve the quality of stallion semen, with respect to sperm morphology and chromatin integrity, both of which have been linked to fertility, using either density gradient centrifugation (DGC) or a new method, hereby named single layer centrifugation (SLC). The two methods of colloidal centrifugation were evaluated using 38 ejaculates from 10 stallions. Sperm morphology, subjective motility and sperm chromatin integrity were compared in uncentrifuged samples and in centrifuged sperm preparations. The proportion of morphologically normal spermatozoa varied between stallions (p < 0.001) and was increased by both methods of colloidal centrifugation (median value before centrifugation 67.5%; after SLC 78%; after DGC 77%; p < 0.001). The incidence of certain abnormalities was reduced, e.g. proximal cytoplasmic droplets were reduced from 12.9% to 8.8% (p < 0.001), and mid-piece defects from 5.3% to 1.4% (p < 0.05). Similarly, sperm motility and chromatin integrity were significantly improved (p < 0.001), with no difference between the two centrifugation methods. Centrifugation through colloids can enrich the proportions of stallion spermatozoa with normal morphology and normal chromatin structure in sperm preparations. The new method, SLC, was as effective as DGC in selecting motile stallion spermatozoa with normal morphology and intact chromatin. SLC, being simpler to use than DGC, would be appropriate for routine use by stud personnel to improve stallion sperm quality in insemination doses.