Martins Muller Carrara, Gonçalves Lucas Melo, Nonato Amanda, Nassif Travençolo Bruno Augusto, Alves Benner Geraldo, Beletti Marcelo Emílio
Laboratory of Biology of Reproduction, Federal University of Uberlândia, MG, Brazil.
Faculty of Computer Science, Federal University of Uberlândia, MG, Brazil.
Theriogenology. 2021 Feb;161:200-209. doi: 10.1016/j.theriogenology.2020.12.004. Epub 2020 Dec 9.
The aim of this study was to evaluate the sperm head morphometry and chromatin condensation at different regions of the reproductive tract in bulls. Sperm smears of seminiferous tubules (ST), epididymis head (EH), body (EB), and tail (ET), and ductus deferens (DD) were stained with toluidine blue. Afterwards, the sperm head morphometry and chromatin alteration types were evaluated by a computational image analysis. Overall, spermatozoa of ST had lower (P < 0.05) area (A), perimeter (P), width (W), length (L), ellipticity (E), and Fourier harmonics (F0, F1, and F2). The chromatin decondensation (CD) and heterogeneity (CH) were higher (P < 0.05) in the ST region and decreased (P < 0.0001) during the migration along the reproductive tract (ST - DD direction). Considering the factors extracted (Factors 1 and 2) by the principal component analysis, the parameters A, P, W, L, and F0 were responsible for ∼36% of the Factor 1, while the E, F0, F1, and anterior-posterior symmetry (APS) contributed ∼27% to Factor 2. Both, CD and CH were associated with Factor 1 in the EH and ET regions and Factor 2 in the ST. Also, a well-defined difference between sperm heads collected from the ST and DD regions was observed by canonical analysis. The distribution of each chromatin alteration type was recorded. The proportion of normal sperm was lower (P < 0.05) in ST compared to other regions. Moreover, the chromatin influenced the morphometry and sperm heads with whole chromatin alteration type showed a smaller (P < 0.05) A, P, W, L, and E. In summary, the epididymal maturation is important for chromatin compaction and final morphometry of the sperm head. Also, the identification and quantification of the sperm chromatin condensation in different regions of reproductive tract can be used as potential biomarkers to predict the fertility in bulls.
本研究的目的是评估公牛生殖道不同区域的精子头部形态测量和染色质凝聚情况。用甲苯胺蓝对生精小管(ST)、附睾头(EH)、附睾体(EB)、附睾尾(ET)以及输精管(DD)的精子涂片进行染色。之后,通过计算机图像分析评估精子头部形态测量和染色质改变类型。总体而言,ST的精子具有较低(P < 0.05)的面积(A)、周长(P)、宽度(W)、长度(L)、椭圆率(E)以及傅里叶谐波(F0、F1和F2)。染色质解凝聚(CD)和异质性(CH)在ST区域较高(P < 0.05),并且在沿生殖道迁移(ST - DD方向)过程中降低(P < 0.0001)。考虑到主成分分析提取的因素(因素1和因素2),参数A、P、W、L和F0对因素1的贡献率约为36%,而E、F0、F1和前后对称性(APS)对因素2的贡献率约为27%。在EH和ET区域,CD和CH均与因素1相关,而在ST区域与因素2相关。此外,通过典型分析观察到从ST和DD区域采集的精子头部之间存在明确差异。记录了每种染色质改变类型的分布情况。与其他区域相比,ST中正常精子的比例较低(P < 0.05)。此外,染色质影响形态测量,具有整体染色质改变类型的精子头部显示出较小(P < 0.05)的A、P、W、L和E。总之,附睾成熟对于染色质压缩和精子头部的最终形态测量很重要。此外,对生殖道不同区域精子染色质凝聚的鉴定和定量可作为预测公牛生育力的潜在生物标志物。