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公猪精子液态保存前去除精浆:一种可提高其受精能力的做法。

Removal of seminal plasma prior to liquid storage of boar spermatozoa: A practice that can improve their fertilizing ability.

作者信息

Pavaneli Ana Paula Pinoti, Passarelli Marina da Silva, de Freitas Flávia Vieira, Ravagnani Gisele Mouro, Torres Mariana Andrade, Martins Simone Maria Massami Kitamura, Yeste Marc, de Andrade André Furugen Cesar

机构信息

Laboratory of Andrology and Technology of Swine Embryos, Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São Paulo, Pirassununga, São Paulo, Brazil.

Biotechnology of Animal and Human Reproduction (TechnoSperm), Unit of Cell Biology, Department of Biology, Institute of Food and Agricultural Technology, Faculty of Sciences, University of Girona, Girona, Spain.

出版信息

Theriogenology. 2019 Feb;125:79-86. doi: 10.1016/j.theriogenology.2018.10.020. Epub 2018 Oct 25.

Abstract

Seminal plasma (SP) plays a vital role in the maintenance of sperm function and integrity along with being involved in their communication with the female reproductive tract. Under in vitro conditions, although it is generally accepted that boar semen is better preserved when SP is diluted (extended) or removed (cryopreserved), its role during storage is not completely elucidated. In this context, the current study sought to determine the role of SP during storage of boar spermatozoa at 17 °C for 72 h. Thus, two treatments were prepared with semen from the sperm-rich fraction (SRF) of boar ejaculate previous to storage in liquid state: 1) PSP: semen directly extended in Beltsville Thawing Solution (BTS), and 2) ASP: semen first centrifuged with subsequent removal of supernatant (containing SP) followed by suspension of sperm in BTS. From this, two experiments were conducted separately in this work: 1) in vitro and 2) in vivo assays. The former aimed to evaluate how sperm capacity responds to in vitro capacitation (IVC) and whether their quality is affected by previous exposure to SP. In the latter, the objective was to understand how important these previous conditions can be for reproductive performance after artificial insemination (AI). According to our results, the previous removal of SP does not affect sperm quality and the response of these cells to IVC (P > 0.05) along with resulting in a lower percentage of acrosome damage in them [12.87 ± 0.76 (ASP) vs. 16.38 ± 0.73 (PSP)] (P < 0.05). This improved preservation of acrosome integrity in the absence of SP can explain the higher fertility rate (%) [63.27 ± 23.47 (ASP) vs. 38.57 ± 16.30 (PSP)] and number of implanted embryos at 28 days after AI (13.71 ± 4.88 (ASP) vs. 7.16 ± 4.02 (PSP)] (P < 0.05) presented by gilts inseminated with seminal doses of ASP. In conclusion, removal of SP prior to liquid storage of boar sperm for 72 h can be beneficial for their fertilizing ability.

摘要

精浆(SP)在维持精子功能和完整性以及参与精子与雌性生殖道的交流方面发挥着至关重要的作用。在体外条件下,尽管人们普遍认为当精浆被稀释(扩充)或去除(冷冻保存)时,公猪精液能得到更好的保存,但其在储存过程中的作用尚未完全阐明。在此背景下,本研究旨在确定精浆在公猪精子于17°C储存72小时期间的作用。因此,在精液以液态储存之前,对公猪射精富含精子部分(SRF)的精液进行了两种处理:1)PSP:精液直接用贝尔茨维尔解冻液(BTS)扩充;2)ASP:精液先离心,随后去除上清液(含精浆),然后将精子悬浮于BTS中。据此,本研究分别进行了两项实验:1)体外实验和2)体内实验。前者旨在评估精子能力对体外获能(IVC)的反应以及其质量是否受先前接触精浆的影响。在后者中,目的是了解这些先前条件对人工授精(AI)后繁殖性能的重要性。根据我们的结果,先前去除精浆不会影响精子质量以及这些细胞对IVC的反应(P>0.05),同时导致精子顶体损伤百分比降低[12.87±0.76(ASP)对16.38±0.73(PSP)](P<0.05)。在不存在精浆的情况下顶体完整性的这种改善保存可以解释用ASP精液剂量授精的后备母猪呈现出的更高受精率(%)[63.27±23.47(ASP)对38.57±16.30(PSP)]以及人工授精后28天植入胚胎的数量(13.71±4.88(ASP)对7.16±4.02(PSP)](P<0.05)。总之,在公猪精子液态储存72小时之前去除精浆对其受精能力可能有益。

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