评估公猪精液质量与生育力的关系。
Assessment of sperm quality traits in relation to fertility in boar semen.
机构信息
Department of Non-infectious diseases, Faculty of Veterinary Medicine, Lithuanian Veterinary Academy, Kaunas, Lithuania.
出版信息
Acta Vet Scand. 2009 Dec 16;51(1):53. doi: 10.1186/1751-0147-51-53.
BACKGROUND
Several studies have been published where sperm plasma membrane integrity correlated to fertility. In this study we describe a simple fluorometer-based assay where we monitored the fluorescence intensity of artificially membrane-ruptured spermatozoa with a fixed time staining with fluorescent DNA dyes.
METHODS
Membrane-impermeant fluorescent dyes Hoechst 33258 (H258) and propidium iodide (PI) were used to measure the fluorescence of the nucleus in artificially membrane ruptured spermatozoa and membrane-permeant dye Hoechst 33342 (H342) was used to measure fluorescence of intact spermatozoa. The concentration of spermatozoa in insemination doses varied from 31.2x10(6)/ml to 50x10(6)/ml and the average value was 35x10(6)/ml. Each boar was represented by three consecutive ejaculates, collected at weekly intervals. Nonreturn rate within 60 days of first insemination (NR%) and litter size (total number of piglets born) of multiparous farrowings were used as fertility measures.
RESULTS
Sperm fluorescence intensity of H258 and H342, but not the fluorescence intensity of PI-stained spermatozoa correlated significantly with the litter size of multiparous farrowings, values being r=-0.68 (P<0.01) for H258, r=-0.69 (P<0.01) for H342 and r=-0.38, (P=0.11) for PI.
CONCLUSIONS
The increase in fluorescence values of membrane-ruptured H258 and unruptured H342-stained spermatozoa in boar AI doses can be associated with smaller litter size after AI. This finding indicates that the fluorescence properties of the sperm nucleus could be used to select for AI doses with greater fertilizing potential.
背景
已有多项研究表明，精子质膜完整性与生育力相关。本研究采用一种简单的基于荧光计的方法，通过固定时间用荧光 DNA 染料对人工质膜破裂的精子进行染色，监测其荧光强度。
方法
采用膜不可渗透的荧光染料 Hoechst 33258（H258）和碘化丙啶（PI）来测量人工质膜破裂精子的核荧光强度，而膜可渗透的染料 Hoechst 33342（H342）则用于测量完整精子的荧光强度。授精剂量中的精子浓度从 31.2x10(6)/ml 到 50x10(6)/ml 不等，平均值为 35x10(6)/ml。每头公猪连续采集三个精液样本，每周采集一次。首次授精后 60 天内的返情率（NR%）和经产窝产仔数（出生仔猪总数）被用作生育力指标。
结果
H258 和 H342 标记的精子荧光强度，但不是 PI 染色精子的荧光强度，与经产窝产仔数显著相关，H258 的 r 值为-0.68（P<0.01），H342 的 r 值为-0.69（P<0.01），PI 的 r 值为-0.38（P=0.11）。
结论
在 AI 剂量中，H258 和未破裂的 H342 标记的精子的荧光值增加，可能与 AI 后窝产仔数减少有关。这一发现表明，精子核的荧光特性可用于选择具有更大受精潜力的 AI 剂量。
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