Fibronectin induces capacitation-associated events through the endocannabinoid system in bull sperm
Fibronectin induces capacitation-associated events through the endocannabinoid system in bull sperm
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DOI：
10.1016/j.theriogenology.2020.04.031
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发表时间：
2020-09-01
期刊：
影响因子：
2.8
通讯作者：
Perez-Martinez, S.
中科院分区：
文献类型：
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作者：
Osycka-Salut, C. E.;Martinez-Leon, E.;Perez-Martinez, S.
哺乳动物射出的精子必须在雌性生殖道中经历一系列变化，统称为获能，才能使卵母细胞受精。我们报道了一种来自细胞外基质的糖蛋白纤维连接蛋白（Fn）和内源性大麻素家族的主要成员之一anandamide （AEA）存在于牛输卵管液中并调节公牛精子的功能。此外，AEA通过CB1和TRPV1受体诱导牛精子获能。在这项工作中，我们研究了Fn是否通过激活内源性大麻素系统来诱导牛精子获能。我们用Fn （100 mg/ml）和（或）capsazepine(一种TRPV1拮抗剂（0.1 μ M）孵育精子，并评估了一些与精子获能相关的事件，如lpc诱导的顶体反应、精子从输卵管中释放、PKA磷酸化底物（pPKAs）的诱导、蛋白酪氨酸磷酸化（pY）和一氧化氮（NO）的产生。此外，我们还研究了脂肪酸酰胺水解酶（FAAH）的活性，这是一种降解AEA的酶。我们发现Fn通过α 5 β 1整合素诱导了与能力相关的事件。此外，Fn刺激了cAMP/PKA和NO/NO合成酶等与获能相关的信号通路。此外，Fn降低了FAAH活性，这与精子获能有关。辣椒平逆转纤维连接蛋白诱导的能化，以及pPKAs和NO水平。用稳定的AEA类似物r -甲烷酰胺（1.4 nM）孵育精子，可提高cAMP和pPKAs水平。H89 （50 μ M）或KT5720 (100 nM) （PKA抑制剂）的存在阻止了aea诱导的能化。此外，r -甲烷酰胺和辣椒素（0.01 mM） （TRPV1激动剂）通过PKA途径增加NO的产生。这些结果表明Fn通过α 5 β 1支持牛精子的能化。这种作用依赖于通过cAMP/PKA和NO信号通路激活TRPV1。我们认为Fn可能是一种促进公牛精子获能的新药物。我们的发现有助于更好地理解Fn信号在包括人类在内的哺乳动物成功受精和胚胎发育的激活事件中的意义。(C) 2020爱思唯尔公司版权所有。
Mammalian ejaculated spermatozoa must undergo a series of changes in the female reproductive tract, collectively called capacitation, in order to fertilize the oocyte. We reported that fibronectin (Fn), a glycoprotein from the extracellular matrix, and anandamide (AEA), one of the major members of the endocannabinoid family, are present in the bovine oviductal fluid and regulate bull sperm function. Also, AEA induces bovine sperm capacitation, through CB1 and TRPV1 receptors.In this work, we investigated if Fn induces bovine sperm capacitation thought the activation of the endocannabinoid system in this process. We incubated sperm with Fn (100 mg/ml) and/or capsazepine, a TRPV1 antagonist (0.1 mu M) and some events related to sperm capacitation such as LPC-induced acrosome reaction, sperm-release from the oviduct, induction of PKA phosphorylated substrates (pPKAs) and protein tyrosine phosphorylation (pY) and nitric oxide (NO) production were assessed. Also, we studied the activity of fatty acid amide hydrolase (FAAH), the enzyme that degrades AEA. We found that Fn, via alpha 5 beta 1 integrin, induced capacitation-associated events. Also, Fn stimulated signaling pathways associated to capacitation as cAMP/PKA and NO/NO synthase. Moreover, Fn decreased the FAAH activity and this correlated with sperm capacitation. Capsazepine reversed fibronectin-induced capacitation, and pPKAs and NO levels. The incubation of spermatozoa with R-methanandamide (1.4 nM), a stable analogue of AEA, increased cAMP and pPKAs levels. The presence of H89 (50 mu M) or KT5720 (100 nM) (PKA inhibitors) prevented AEA-induced capacitation. In addition, R-methanandamide and capsaicin (0.01 mM), a TRPV1 agonist, increased NO production via the PKA pathway. These results indicate that Fn, through alpha 5 beta 1, supports capacitation in bovine spermatozoa. This effect is dependent on the activation of TRPV1 through cAMP/PKA and NO signaling pathways. We propose that Fn could be considered as a new agent that promotes sperm capacitation in bull sperm. Our findings contribute to better understand the significance of Fn signaling in the capacitating events that lead to successful fertilization and embryo development in mammals including humans. (C) 2020 Elsevier Inc. All rights reserved.