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新鲜及冻融隼类射精精液中未成熟和成熟精子的形态测定

Immature and mature sperm morphometry in fresh and frozen-thawed falcon ejaculates.

作者信息

Villaverde-Morcillo S, Soler A J, Esteso M C, Castaño C, Miñano-Berna A, Gonzalez F, Santiago-Moreno J

机构信息

Veterinary Clinical and Research Services, Fieb Foundation, Madrid, Spain.

SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario, Albacete, Spain.

出版信息

Theriogenology. 2017 Aug;98:94-100. doi: 10.1016/j.theriogenology.2017.04.051. Epub 2017 May 4.

Abstract

Sperm morphometry is one characteristic which may be useful in prediction of fertility and sperm freezability in a species. Knowledge of the sperm characteristics of the ejaculate and the morphometric descriptors is necessary to effectively develop sperm cryopreservation. The aim of the current study was to provide a general description of the sperm from two falcon species (Peregrine falcon Falco peregrinus peregrinus/brookei and Gyrfalcon Falco rusticolus) including immature sperm, sperm head morphometric descriptors, and the existence of mature sperm subpopulations. Semen samples were collected by massage and voluntary false copulation and diluted with Lake and Ravie medium. Smears were prepared of the diluted samples, stained with Hemacolor, and subjected to: 1) morphological analysis (bright field optical microscopy), and 2) computerised morphometric analysis; each sperm head was measured for length, width, area and perimeter. In addition, in the Gyrfalcon, pooled semen was frozen in pellets using DMA as a cryoprotectant and the analyses repeated after thawing. The mean percentage of immature sperm (spermatocytes and spermatids) was similarly high in all species/subspecies: Brookei Peregrine falcon (F. p. brookei) 55.5%, European Peregrine falcon (F. p. peregrinus) 65.5% and Gyrfalcon 64.7%. Clustering analyses identified four subpopulations of mature spermatozoa with different morphometric characteristics (P < 0.001). The relative proportions of these subpopulations were similar in all three species. The mean values recorded for the morphometric variables of the four subpopulations were, however, lower (P < 0.001) in the thawed Gyrfalcon samples than in fresh samples. The results support the idea of pleiomorphy as a characteristic of raptor mature sperm. This finding, plus that of the existence of four sperm subpopulations with different morphometric characteristics, may be important in the future development of cryopreservation protocols for falcon sperm.

摘要

精子形态测定是一个可能有助于预测某一物种生育能力和精子冷冻保存能力的特征。了解射精的精子特征和形态学描述符对于有效开发精子冷冻保存技术是必要的。本研究的目的是对两种隼类(矛隼Falco peregrinus peregrinus/brookei和矛隼Falco rusticolus)的精子进行一般性描述,包括未成熟精子、精子头部形态学描述符以及成熟精子亚群的存在情况。通过按摩和自愿假交配收集精液样本,并用Lake和Ravie培养基稀释。对稀释后的样本制备涂片,用Hemacolor染色,并进行:1)形态学分析(明场光学显微镜),以及2)计算机形态测定分析;测量每个精子头部的长度、宽度、面积和周长。此外,对于矛隼,将混合精液用二甲基亚砜作为冷冻保护剂制成颗粒冷冻,并在解冻后重复进行分析。所有物种/亚种中未成熟精子(精母细胞和精子细胞)的平均百分比同样很高:布鲁克矛隼(F. p. brookei)为55.5%,欧洲矛隼(F. p. peregrinus)为65.5%,矛隼为64.7%。聚类分析确定了具有不同形态特征的四个成熟精子亚群(P < 0.001)。这三个物种中这些亚群的相对比例相似。然而,解冻后的矛隼样本中四个亚群的形态学变量记录的平均值低于新鲜样本(P < 0.001)。结果支持多形性是猛禽成熟精子特征的观点。这一发现,加上存在四个具有不同形态特征的精子亚群这一发现,可能对未来隼类精子冷冻保存方案的开发很重要。

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