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用于表征冷冻保存的牛精子经Percoll梯度离心后顶体损伤的透射电子显微镜检查。

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa.

作者信息

Oliveira Letícia Z, Hossepian de Lima Vera Fm, Levenhagen Marcelo A, Santos Ricarda M, Assumpção Terezinha I, Jacomini José O, Andrade André Fc, Arruda Rubens P, Beletti Marcelo E

机构信息

Department of Preventive Veterinary Medicine and Animal Reproduction, School of Agrarian Sciences and Veterinary Medicine, Universidade Estadual Paulista, Jaboticabal 14884-900, Brazil.

出版信息

J Vet Sci. 2011 Sep;12(3):267-72. doi: 10.4142/jvs.2011.12.3.267.

DOI:10.4142/jvs.2011.12.3.267
PMID:21897100
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3165156/
Abstract

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

摘要

本研究的目的是对冷冻保存的牛精子经不连续 Percoll 梯度离心后的顶体超微结构进行表征。从六个不同品种的公牛采集精液，每头公牛评估三份射精样本。将冷冻精液样本解冻，在 Percoll 离心前（n = 18）和离心后（n = 18）通过透射电子显微镜（TEM）评估精子细胞的顶体区域。对所分析的 36 个样本中的每个样本的 20 个精子头部进行评估，确保研究了大量细胞。数据在 5% 的显著性水平下进行方差分析。Percoll 离心降低了具有正常顶体的精子百分比（从 61.77% 降至 30.24%），降低了呈现非典型顶体反应的精子百分比（从 28.38% 降至 4.84%），并增加了顶体受损的精子百分比（从 6.14% 增至 64.26%）。具有典型顶体反应的精子百分比在离心前（3.70%）和离心后（0.67%）没有显著差异。TEM 区分了四种不同类型的顶体状态，并能够对顶体损伤进行超微结构表征。在所采用的 Percoll 梯度离心方案中，具有正常顶体的精子百分比下降，顶体损伤是最常见的顶体损伤类型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/951b/3165156/2c6df5fe8f41/jvs-12-267-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/951b/3165156/2c6df5fe8f41/jvs-12-267-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/951b/3165156/2c6df5fe8f41/jvs-12-267-g001.jpg

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