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在精子细胞发生顶体反应并使卵母细胞受精之前,获能是一个重要的生理先决条件。获能是精子在雌性生殖道中进一步成熟的复杂现象,其赋予精子以增强活性的能力,使精子能够与卵母细胞透明带(ZP)相互作用,进行顶体反应并与卵母细胞质膜融合,进而完成受精过程。然而精子获能的分子机制十分复杂,目前还未完全明确,但获能后的精子会有诸多结构及生化方面的变化,如蛋白酪氨酸磷酸化、精子膜胆固醇外流、活性氧的产生及精子膜超极化。蛋白质通过磷酸化或去磷酸化调节精子获能和顶体反应等一些重要的现象,这是精子到达、结合、穿透和融合卵母细胞所必需的过程。因此蛋白磷酸化是获能的一个非常重要的过程,尤其是在酪氨酸残基处的磷酸化是获能过程中发生的最重要的事件之一,且酪氨酸磷酸化可能是细胞中信号转导途径的主要甚至是唯一的指标。作者主要针对蛋白磷酸化、精子中酪氨酸磷酸化的发现、作用和定位,以及影响获能过程中酪氨酸磷酸化的几个因素进行阐述。 相似文献
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哺乳动物精子获能的分子机制 总被引：2，自引：0，他引：2
精子获能是精子能够与卵母细胞发生顶体反应和受精的一个重要生理前提。精子获能的分子机制相当复杂,许多报道表明精子获能受到多种细胞信号途径的调控。尽管目前尚未完全明确,但是许多研究表明获能精子发生许多结构和生化变化,包括蛋白酪氨酸磷酸化、精子膜胆固醇外流、活性氧的产生及精子膜超极化,这些变化都有助于精子获能的发生。Ca2 和HCO3-通过对cAMP的调控有助于获能完成,葡萄糖、孕酮和肝素作为获能液的重要添加物,通过不同途径促发精子获能。文章从这些方面对获能做一综述,在此基础上提出以后的研究方向。 相似文献
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精子获能过程中蛋白酪氨酸磷酸化的研究进展 总被引：1，自引：0，他引：1
精子获能是精子发生顶体反应以及精卵结合受精前重要的生理过程,许多因素参与调节精子获能,其中精子蛋白酪氨酸磷酸化是精子运动力、精子超激活运动、精子获能等多种生理生化过程的重要调控因素。本文就目前有关精子蛋白酪氨酸磷酸化与精子获能之间的研究进展进行综述。 相似文献
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哺乳动物精子与卵细胞结合完成受精前,获能是关键环节之一。精子获能伴随着许多生理生化过程的改变,包括离子含量的改变、质膜重组、胆固醇外排和蛋白质的酪氨酸磷酸化等,此外,去能因子能使精子顺利完成受精。通过这些变化可以检测精子的获能状态,常用的检测方法主要有s ACcAMP-PKA通路检测、质膜流动性变化检测、胆固醇再次分布检测、酪氨酸磷酸化检测和顶体反应检测等。对哺乳动物精子获能过程中涉及的信号通路、信号分子、调节因子、离子通道以及精子获能检测方法的研究进展进行综述,并对未来主要研究方向进行展望,以期为精子体外培养及辅助生殖技术的应用等提供参考。 相似文献
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《经济动物学报》2013,(3)
蛋白的酪氨酸磷酸化对精子运动力的维持,精子获能,超激活运动以及顶体反应等生理过程十分重要。为探讨塔里木马鹿不同生理状态下,不同个体精液精子蛋白酪氨酸磷酸化的关系,以塔里木马鹿冷冻保存的精子为试验材料,采用非离子型去污剂(NP-40)裂解法提取精子膜蛋白,用SDS-PAGE凝胶电泳和Western免疫印迹技术检测精子膜蛋白及发生酪氨酸磷酸化蛋白的表达情况。结果表明:精液异常(黄棕色,黏稠)个体的冻融精子活力,精子蛋白酪氨酸磷酸化水平明显下降(P<0.01);含卵黄稀释液作为冷冻保存稀释时,其冻融精子蛋白的磷酸化水平显著高于无卵黄稀释液的其它个体(P<0.05);另外,相同生理状态下冷冻保存的不同个体的冻融精子酪氨酸磷酸化蛋白的表达水平也有显著差异(P<0.05),但除了精液异常个体外所有公鹿冻融精子的40 ku和47 ku蛋白的磷酸化表现出较强的表达(P<0.05)。以上结果提示,塔里木马鹿冻融精子蛋白酪氨酸磷酸化水平与精液的生理状态有关,并且不同个体之间也存在一定的差异。 相似文献
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第三节受精、妊娠与分娩一、受精(一)配子的运行1.射精部位在交配过程中,公猪将精液直接射到母猪子宫或子宫颈中。母猪的子宫颈没有阴道部,使得精液能顺利射入子宫内。2.精子获能精子在受精前必须在雌性生殖道内经历一段时间,并发生一系列生理性、机能性变化,才具有与卵母细胞受精的能力,这种现象称精子获能。公猪精子获能需要3～6小时。3.受精部位在输卵管上1/3的壶腹部受精。4.顶体反应精子获能之后,在穿越透明带前后,精子顶体开始膨大,精子质膜和顶体外膜开始融合,使精子顶体形成许多泡状结构,通过空泡间隙释放出透明质酸酶、放射冠穿透… 相似文献
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本研究对猪精子获能前后细胞亚组分蛋白进行分离以及对酪氨酸磷酸化蛋白进行鉴定,旨在为哺乳动物精子受精生物学研究奠定理论基础。利用动物精子体外获能培养、细胞亚组分分离技术及蛋白免疫印迹的方法,分离猪精子细胞亚组分蛋白及酪氨酸磷酸化蛋白鉴定。结果表明,猪精子经过获能培养后各项活力指标均得到显著提高,且与精子蛋白发生酪氨酸磷酸化修饰密切相关;获能精子中126、108、79ku的高分子量蛋白磷酸化程度明显高于未获能精子;分子质量约为25、47、50ku的膜蛋白及47ku胞浆蛋白发生酪氨酸磷酸化,其中25、47ku的膜蛋白酪氨酸磷酸化程度显著高于未获能精子(P<0.05);分子量约为23、37、42～50ku的核蛋白发生酪氨酸磷酸化,获能精子中23ku的核蛋白酪氨酸磷酸化程度显著高于未获能精子(P<0.05)。结果提示,猪精子细胞不同亚组分中,发生酪氨酸磷酸化修饰的蛋白以膜蛋白及核蛋白为主,同时有少量的胞浆蛋白。 相似文献
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《中国畜牧杂志》2021,(9)
实验旨在分析17℃保存对获能精子蛋白酪氨酸磷酸化的影响,通过精子蛋白亚组分分离及酪氨酸磷酸化鉴定,研究精浆以及中药水提液对保存精子获能相关蛋白酪氨酸磷酸化的影响。选择获能指标-酪氨酸磷酸化分析新鲜猪精子和17℃保存猪精子,提取精子全蛋白、膜蛋白、核蛋白以及骨架蛋白,利用WB分离及鉴定酪氨酸磷酸化差异。结果显示:17℃保存条件下,获能精子在40～45、46～50 ku分子量膜蛋白酪氨酸磷酸化程度减弱;未获能精子在30～35 ku分子量胞浆蛋白磷酸化程度增强,出现似"获能"现象;精浆在保存中起到提供营养的作用但易导致细菌滋生;中药水提液对精子保存过程中的保护效果体现在精子获能培养后,鞭毛中段、主段酪氨酸磷酸化和新鲜精子无明显差异。 相似文献
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Capacitation is a biological phenomenon occurring prior to fertilization and is a multiple event process. Many physiological and biochemical changes takes place during the process; these changes are related to lipid composition of membrane, intracellular modulation of ion concentration, protein phosphorylation, sperm movement and membrane permeability. These events occur when the sperm is exposed to the new environment of ion concentration in the female reproductive tract. Ions such as bicarbonate and calcium facilitate capacitation by activating adenylyl cyclase, thus initiating protein kinase A (PKA) signalling cascade. Extracellular‐regulated kinase pathway is activated by ligand binding to the membrane receptors and intracellular activation by reactive oxygen species (ROS). Activation of these pathways leads to the phosphorylation of different proteins, which is associated with events such as capacitation, hyperactivation and acrosome reaction that are essential for successful fertilization. Extensive studies were carried out on protein phosphorylation in relation to capacitation, but its role still remains ambiguous. 相似文献
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To acquire the fertilizing competence, spermatozoa must undergo a cascade of physiological and biochemical changes collectively defined as capacitation. Compelling evidence signifies that the global increase in protein tyrosine phosphorylation is the driving factor for capacitation. In our laboratory, we previously demonstrated that nitric oxide (NO) induces capacitation in buffalo sperm and is associated with an increase in protein tyrosine phosphorylation. The aim of the present study is to identify the proteins undergo tyrosine phosphorylation during NO induced buffalo sperm capacitation using 2-D immunoblotting and mass spectrometry. The percentage of progressively motile and capacitated sperm was more in presence of l-arginine. Along with known tyrosine phosphoproteins like ATP synthase subunit beta, pyruvate dehydrogenase E1 component subunit beta, GST mu 3, F-actin capping protein subunit beta 2, GPD2 and VDAC2, interestingly novel tyrosine phosphoprotein substrates such as actin, serine/threonine-protein phosphatase PP1-gamma catalytic subunit, and glutamine synthetase were also identified which might be specific to the NO induced signaling and also emphasizes the species specificity with respect to tyrosine phosphorylation of proteins during capacitation. In conclusion, this study forms an essential step in delineating the proteins undergo tyrosine phosphorylation in response to NO induced signaling pathways during capacitation of buffalo sperm. 相似文献
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Mirshokraei P Hassanpour H Mehdizadeh A Akhavan Taheri M 《Research in veterinary science》2011,91(2):281-284
To evaluate effects of different concentrations of pentoxifylline, as phosphodiesterase inhibitor, on quality of motility, capacitation and acrosome reaction, Ejaculated spermatozoa were collected from crossbred dogs. The sperm were incubated at concentrations of 0.1, 1, 10 and 100 mM pentoxifylline for 2 h. Conventional assessment was also made on the percentage of motility and quality of motility of spermatozoa; values were expressed as sperm motility index (SMI). Capacitation and acrosome reaction were also evaluated by chlortetracycline fluorescence staining. SMI as quality index of sperm was significantly increased in concentrations of 10 and 100 mM pentoxifylline during 1 and 2 h compared to control. The number of capacitated or acrosome reacted spermatozoa significantly (P < 0.05) were higher than controls at high concentrations of pentoxifylline (10 and 100 mM) during 1 and 2 h. In conclusion, high concentration of pentoxifylline is able to induce capacitation and acrosome reaction and improves quality of motility in canine ejaculated spermatozoa. 相似文献
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The aim of this research was to study the effect of hyaluronic acid on bovine cryopreserved spermatozoa compared with heparin as regards the variation of capacitation induction, cellular oxidative metabolism and intracellular signal induced by membrane‐associated adenylate cyclase to propose hyaluronic acid as a capacitation inductor. Heparin or hyaluronic acid and lysophosphatidylcholine were used to induce sperm capacitation and acrosome reaction, respectively. 2′,5′‐dideoxyadenosine was used as a membrane‐associated adenylate cyclase inhibitor. The highest percentages of capacitated spermatozoa and live spermatozoa with acrosome integrity were obtained by incubating sperm for 60 min using 1000 μg/ml hyaluronic acid. In these conditions, capacitation induced by hyaluronic acid was lower compared with heparin; nonetheless both glycosaminoglycans promote intracellular changes that allow true acrosome reaction in vitro induced by lysophosphatidylcholine in bovine spermatozoa. Oxygen consumption in heparin‐capacitated spermatozoa was significantly higher than in hyaluronic acid‐treated spermatozoa. With all treatments, mitochondrial coupling was observed when a specific uncoupler of the respiratory chain was added. The inhibition of membrane‐associated adenylate cyclase significantly blocked capacitation induction produced by hyaluronic acid, maintaining a basal sperm oxygen uptake in contrast to heparin effect in which both sperm parameters were inhibited, suggesting that the membrane‐associated adenylate cyclase activation is involved in the intracellular signal mechanisms induced by both capacitation inductors, but only regulates mitochondrial oxidative phosphorylation in heparin‐capacitated spermatozoa. 相似文献
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Choi YJ Uhm SJ Song SJ Song H Park JK Kim T Park C Kim JH 《The Journal of reproduction and development》2008,54(1):68-83
To identify the mechanisms underlying capacitation, we undertook a high-resolution differential proteomic analysis of pig sperm cells. Two-dimensional gel electrophoresis and subsequent MALDI-TOF mass spectrometry analyses led to identification of 56 differentially expressed proteins. After induction of capacitation in vitro, the well-established markers of the capacitation (lactadherin P47, acrosomal protein SP-10 precursor, prohibitin, proteasomes, DJ-1 protein and arylsulfatase-A) and TCA cycle proteins (isocitrate dehydrogenase, malate dehydrogenase and pyruvate dehydrogenase) were identified. During induction, cytochrome c expression via the p53 pathway increased, however apoptotic executors, such as caspase-3, decreased significantly. Therefore, we tested the hypothesis that cytochrome c upregulation in spermatozoa is capable of activating tyrosine phosphorylation for capacitation, rather than apoptosis. Exposure of sperm cells to soluble Na2CrO4 [Cr (VI)], which induces cytochrome c upregulation, caused a dose- and time-dependent increase in tyrosine phosphorylation of sperm proteins in non-capacitating medium. In contrast, supplementation of cyclosporin A, which blocks cytochrome c upregulation, inhibited tyrosine phosphorylation of sperm proteins. Furthermore, spermatozoa in capacitation medium or non-capacitation media supplemented with soluble Cr (VI) showed similar levels of capacitation. These findings indicate that differential expression of many of these proteins has previously been unrecognized in sperm cells incubated in capacitation medium also suggest that a gradual increase of cytochrome c during incubation to induce capacitation determines sperm cell fate, i.e., apoptosis or further development for fertilization. 相似文献
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Participation of phosphofructokinase,malate dehydrogenase and isocitrate dehydrogenase in capacitation and acrosome reaction of boar spermatozoa 
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下载免费PDF全文 E Breininger D Dubois VE Pereyra PC Rodriguez MM Satorre PD Cetica 《Reproduction in domestic animals》2017,52(5):731-740
The aim of this work was to determine the enzymatic activity of phosphofructokinase (PFK), malate dehydrogenase (MDH) and isocitrate dehydrogenase (IDH) in boar spermatozoa and study their participation in bicarbonate‐induced capacitation and follicular fluid‐induced acrosome reaction. Enzymatic activity of these enzymes was determined spectrophotometrically in extracts of boar spermatozoa. Sperm suspensions were incubated in the presence of bicarbonate (40 mM), a well‐known capacitation inducer, or follicular fluid (30%), as an acrosome reaction inducer, and different concentrations of oxoglutarate, oxalomalate and hydroxymalonate, inhibitors of PFK, IDH and MDH, respectively. Capacitation percentages were determined by the fluorescence technique of chlortetracycline (CTC), and true acrosome reaction was determined by trypan blue and differential–interferential contrast, optical microscopy. The activity of PFK in boar spermatozoa enzymatic extracts was 1.70 ± 0.19 U/1010 spermatozoa, the activity of NAD‐ and NADP‐dependent IDH was 0.111 ± 0.005 U/1010 and 2.22 ± 0.14 U/1010 spermatozoa, respectively, and the activity of MDH was 4.24 ± 0.38 U/1010 spermatozoa. The addition of the specific inhibitors of these enzymes prevented sperm capacitation and decreased sperm motility during capacitation and inhibited the acrosome reaction (AR), without affecting the sperm motility during this process. Our results demonstrate the participation of PFK, IDH and MDH in bicarbonate‐induced capacitation and follicular fluid‐induced acrosome reaction in boar spermatozoa, contributing to elucidate the mechanisms that produce energy necessary for these processes in porcine spermatozoa. 相似文献