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添加外源性孕酮和孕激素受体抑制剂(RU486)对猪冷冻精液稀释液的影响。

Effect of the addition of exogenous progesterone and the progesterone receptor inhibitor (RU 486) on boar cryopreservation semen extenders.

机构信息

Department of Animal Reproduction, Spanish National Institute for Agricultural and Food Research and Technology (INIA-CSIC), 28040 Madrid, Spain.

Department of Animal Reproduction, Spanish National Institute for Agricultural and Food Research and Technology (INIA-CSIC), 28040 Madrid, Spain.

出版信息

Res Vet Sci. 2024 Nov;180:105400. doi: 10.1016/j.rvsc.2024.105400. Epub 2024 Aug 30.

Abstract

Cryopreservation of porcine spermatozoa is detrimental due to their high sensitivity to cold shock, leading to changes akin to capacitation, known as cryocapacitation. These changes, including the acrosomal reaction, hypermotility induction, and protein phosphorylation, might be influenced by the presence of progesterone in seminal plasma and egg yolk, used in most freezing extenders. We tested the effect of various progesterone concentrations added to the freezing extenders (1, 10, and 100 μg/mL). At 100 μg/mL, progesterone decreased the proportion of straightness and tended to reduce viability and the proportion of progressive motility (p < 0.1). At 10 μg/mL, it increased reacted acrosomes in dead sperm (p < 0.05), protein phosphorylation rate (p < 0.05), and tended (p < 0.1) to enhance linear movement compared to the control. To counteract the capacitating effect of progesterone, we examined the effect of antiprogesterone mifepristone (RU 486) at concentrations of 5, 10, 20, 50, 100, and 200 μM, and co-incubated 10 μM of RU 486 with 10 μg/mL of progesterone. RU 486 maintained capacitation levels and motility parameters similar to the control, although high concentrations (100 μM) tended (p = 0.152) to increase protein phosphorylation. Co-incubation reduced the acrosome reaction in dead sperm, and RU 486 appeared to prevent hypermotility stabilizing motility and viability parameters compared to samples with progesterone alone. Protein phosphorylation increased and RU 486 could not restore capacitation to control levels due to its competitive antagonism for progesterone receptors, having less affinity than progesterone, which displaces RU 486 at high concentrations, allowing normal sperm capacitation.

摘要

猪精子的冷冻保存是有害的,因为它们对冷休克非常敏感,导致类似于顶体反应的变化,称为冷冻诱导顶体反应。这些变化包括顶体反应、超激活诱导和蛋白质磷酸化,可能受到精液和蛋黄中孕酮的影响,这是大多数冷冻液中常用的。我们测试了在冷冻液中添加不同浓度的孕酮(1、10 和 100μg/ml)的效果。在 100μg/ml 时,孕酮降低了直线速度的比例,并且倾向于降低活力和直线运动的比例(p<0.1)。在 10μg/ml 时,它增加了死精子中的反应顶体(p<0.05)、蛋白质磷酸化率(p<0.05),并与对照组相比,直线运动趋势增强(p<0.1)。为了抵消孕酮的顶体激活作用,我们检查了抗孕酮米非司酮(RU 486)在 5、10、20、50、100 和 200μM 浓度下的效果,并将 10μM RU 486 与 10μg/ml 孕酮共孵育。RU 486 维持与对照组相似的顶体激活水平和运动参数,尽管高浓度(100μM)有增加的趋势(p=0.152),增加蛋白质磷酸化。共孵育降低了死精子中的顶体反应,与单独使用孕酮的样品相比,RU 486 似乎通过稳定运动和活力参数来防止超激活,从而稳定活力和活力参数。由于 RU 486 对孕酮受体具有竞争性拮抗作用,因此蛋白质磷酸化增加,RU 486 不能将顶体激活恢复到对照水平,因为其对孕酮受体的亲和力低于孕酮,在高浓度时会取代 RU 486,从而允许正常的精子顶体激活。

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