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精浆影响原驼精子的存活率和运动模式。

Seminal plasma affects the survival rate and motility pattern of raw llama spermatozoa.

作者信息

Fumuso F G, Giuliano S M, Chaves M G, Neild D M, Miragaya M H, Gambarotta M C, Carretero M I

机构信息

Cátedra de Teriogenología, Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Buenos Aires, Argentina.

Cátedra de Física Biológica, Universidad de Buenos Aires, Facultad de Ciencias Veterinarias, Instituto de Investigación y Tecnología en Reproducción Animal, Buenos Aires, Argentina.

出版信息

Anim Reprod Sci. 2018 May;192:99-106. doi: 10.1016/j.anireprosci.2018.02.019. Epub 2018 Feb 24.

Abstract

UNLABELLED

The objectives of this study were to evaluate the effect over time of different percentages of seminal plasma (SP) on llama sperm characteristics in raw semen and correlate the techniques routinely used to evaluate sperm viability and acrosome status with the Fluorescein Isothiocyanate -Arachis hypogea agglutinin/Propidium Iodide (FITC-PNA/PI). Eighteen ejaculates, obtained from 6 male llamas using electroejaculation, were incubated in 0.1% collagenase in HEPES-TALP (HT), centrifuged and resuspended with SP and HT: 0, 10, 50 and 100% SP. Samples were incubated (37 °C) until evaluation at 0; 1.5 and 3 h. Split plot and factorial designs were used to analyze sperm motility, viability, membrane function and acrosome status and Spearman's test was used for correlation. At 0 h, samples with 100% SP showed oscillatory motility; whereas in samples with 0 and 10% SP, progressive motility was predominant. Viability, membrane function and total motility decreased significantly at 3 h of incubation in samples with 100% SP. Sperm with intact acrosomes were fewer in 0% SP media at all times. FITC-PNA/PI correlated with 6-Carboxyfluorescein Diacetate and Propidium Iodide (CFDA/PI) and with Coomassie Blue (CB) stains (r = 0.8; p = 0.0 and r = 0.5; p = 0.0 respectively).

CONCLUSIONS

the motility pattern of llama sperm is influenced by the concentration of SP. The use of SP as the only medium is not able to maintain sperm motility, viability and membrane function for 3 h. A certain percentage of SP is necessary in the medium to avoid spontaneous acrosome reactions. The correlations observed could help to shorten evaluation times and reduce costs in sperm laboratories.

摘要

未标记

本研究的目的是评估不同百分比的精浆(SP)对原精液中美洲驼精子特性随时间的影响,并将常规用于评估精子活力和顶体状态的技术与异硫氰酸荧光素-花生凝集素/碘化丙啶(FITC-PNA/PI)进行关联。从6只雄性美洲驼通过电刺激采精获得18份射精样本,在含有0.1%胶原酶的HEPES-TALP(HT)中孵育,离心后用精浆和HT重悬:0、10、50和100%精浆。样本在37°C孵育,直至在0、1.5和3小时进行评估。采用裂区设计和析因设计分析精子活力、生存力、膜功能和顶体状态,并使用Spearman检验进行相关性分析。在0小时时,含有100%精浆的样本显示出摆动性运动;而在含有0%和10%精浆的样本中,主要是进行性运动。在含有100%精浆的样本中孵育3小时后,生存力、膜功能和总活力显著下降。在所有时间点,0%精浆培养基中具有完整顶体的精子较少。FITC-PNA/PI与6-羧基荧光素二乙酸酯和碘化丙啶(CFDA/PI)以及考马斯亮蓝(CB)染色相关(r分别为0.8;p = 0.0和r = 0.5;p = 0.0)。

结论

美洲驼精子的运动模式受精浆浓度影响。仅使用精浆作为培养基无法维持精子活力、生存力和膜功能3小时。培养基中需要一定比例的精浆以避免自发顶体反应。观察到的相关性有助于缩短精子实验室的评估时间并降低成本。

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