Effect of heparin on in vitro capacitation of boar sperm
Effect of heparin on in vitro capacitation of boar sperm
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DOI：
10.4067/s0716-97602006000500006
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发表时间：
2006-01-01
影响因子：
6.7
通讯作者：
CABADA, MARCELO O
中科院分区：
文献类型：
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作者：
DAPINO, DORA G;MARINI, PATRICIA E;CABADA, MARCELO O
采用金霉素（CTC）荧光图谱、10 μ M钙离子载体A23187处理精子后发生顶体反应（AR）的能力和精子活力测定等方法，研究了硫酸化糖胺聚糖肝素对猪精子体外获能的影响。精子在补充有10 mM肝素的获能培养基（CM）中孵育长达120 min，显示获能精子（13模式）和顶体反应精子（AR模式）数量增加，而不影响其活力。在这种条件下，将精子在去除白蛋白、钙、碳酸氢盐或其组合的CM中孵育。在肝素存在下。在无钙或无碳酸氢盐培养基中，获能仅为基础，在肝素存在下未显示变化。在不存在白蛋白的情况下，钙和碳酸氢盐的存在刺激获能，通过加入肝素进一步增加获能。这些结果表明，肝素仅在获能条件下增强猪精子的体外获能。此外，当精子与100 μ g/ml生物素化肝素在存在或不存在未标记肝素的情况下孵育时，我们观察到肝素结合位点主要位于猪精子的顶体区域上，以特异性和可饱和的方式。本工作中描述的肝素的体外作用表明，通常存在于雌性生殖道中的硫酸化糖胺聚糖可能在猪的受精过程中发挥重要作用。
Chlortetracycline (CTC) fluorescent pattern, the ability to undergo acrosome reaction (AR) upon exposure to 10 mu M calcium ionophore A23187 and vitality estimation were used to investigate the effect of the sulfated glycosaminoglycan heparin on the in vitro capacitation of porcine spermatozoa. Sperm incubation in capacitating medium (CM) supplemented with 10 mM heparin for up to 120 min, showed an increase in the number of capacitated sperm (13 pattern) and acrosome reacted sperm (AR pattern), without affecting their viability. In this condition, spermatozoa were incubated in CM depleted of albumin, calcium, bicarbonate or combinations. in the presence of heparin. In either calcium or bicarbonate-free media, capacitation was only basal and did not show variations in the presence of heparin. In absence of albumin the presence of calcium and bicarbonate stimulated capacitation, which was further increased by the addition of heparin. These results suggest that heparin enhances in vitro capacitation of porcine sperm only under capacitating conditions. Additionally, when sperm were incubated with 100 mu g/ml biotinylated heparin in the presence or absence of unlabeled heparin, we observed that heparin binding sites were located mostly on the acrosomal region of boar sperm in an specific and saturable manner. The in vitro effect of heparin described in this work indicates that sulfated glycosaminoglycans, which are normally present in the female reproductive tract, might play an important role in the fertilization process in porcines.