Complex CatSper-dependent and independent [Ca2+]i signalling in human spermatozoa induced by follicular fluid.
Complex CatSper-dependent and independent [Ca2+]i signalling in human spermatozoa induced by follicular fluid.
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DOI：
10.1093/humrep/dex269
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发表时间：
2017-10-01
期刊：
影响因子：
--
通讯作者：
Da Silva SM
中科院分区：
文献类型：
--
作者：
Brown SG;Costello S;Kelly MC;Ramalingam M;Drew E;Publicover SJ;Barratt CLR;Da Silva SM
人类卵泡液（hFF）中的孕酮是否激活CatSper，hFF的其他组分是否调节这种效应和/或单独促进hFF诱导的Ca 2+信号传导？hFF有效地刺激CatSper并增加[Ca 2 +]i，主要是由于高浓度的孕酮，然而，hFF的其他组分也有助于[Ca 2 +]i信号传导，包括CatSper通道活性的调节和[Ca 2 +]i振荡的抑制。CatSper是精子中主要的钙离子通道，对孕酮敏感，对生育力至关重要。hFF和hFF中存在的孕酮都影响精子功能并增加其[Ca 2 +]i。这项基础医学研究使用了来自>40名供体的精液样本和来自>50名接受IVF/ICSI手术取卵的患者的hFF。根据苏格兰东部研究伦理服务REC 1的当地伦理批准（13/ES/0091）招募精液供体和患者。采用膜片钳电生理技术检测CatSper和KSper的活性。在精子群体和单细胞中检查精子[Ca 2 +]i反应。计算机辅助精子分析（CASA）参数和穿透粘性介质用于评估功能效应。hFF和孕酮显著增强CatSper电流。在准生理条件下，hFF（高达50%）未能改变膜K+电导或电流逆转电位。hFF和孕酮（在相同浓度下）在精子群体和单细胞中刺激类似的双相[Ca 2 +]i信号。在高hFF浓度（10%）下，[Ca 2 +]i信号的持续（平台）分量始终大于单独孕酮诱导的分量。在单细胞记录中，1%hFF诱导的[Ca 2 +]i振荡类似于孕酮，但10%hFF产生的[Ca 2 +]i振荡被抑制。处理后的'剥离'脂质衍生的介质，hFF未能显着刺激CatSper电流，但诱导小[Ca 2 +]i的反应，大于剥离后的等效浓度的孕酮诱导的。当用3 μM孕酮预处理的精子（以脱敏孕酮反应）用hFF或剥离的hFF刺激时，观察到类似的[Ca 2 +]i反应。hFF刺激粘性介质渗透，并且比等效剂量的孕酮更有效。N/A.这是一项体外研究。在体内外推这些结果时必须谨慎。这项研究直接证明了hFF激活CatSper，并确定hFF的生物学重要作用至少部分反映了主要通过孕酮对该通道的作用。然而，这些实验也表明，hFF的其他组分都有助于[Ca 2 +]i信号和调节CatSper的激活。在复杂的体内环境之外进行的简单体外实验需要谨慎解释。资金由MRC提供（MR/K 013343/1，MR/012492/1）（S.G.B.，SJP C.L.R.B.）和阿伯泰大学（S.G.B.休假）。额外的资金由TENOVUS SCOTLAND（S.M.D.S.）提供，首席科学家办公室/NHS苏格兰研究（S.M.D.S）。C.L.R.B.是MHR的EIC和WHO男性不育诊断ESG的主席。其余作者没有利益冲突。
Does progesterone in human follicular fluid (hFF) activate CatSper and do other components of hFF modulate this effect and/or contribute separately to hFF-induced Ca2+ signaling? hFF potently stimulates CatSper and increases [Ca2+]i, primarily due to high concentrations of progesterone, however, other components of hFF also contribute to [Ca2+]i signaling, including modulation of CatSper channel activity and inhibition of [Ca2+]i oscillations. CatSper, the principal Ca2+ channel in spermatozoa, is progesterone-sensitive and essential for fertility. Both hFF and progesterone, which is present in hFF, influence sperm function and increase their [Ca2+]i. This basic medical research study used semen samples from >40 donors and hFF from >50 patients who were undergoing surgical oocyte retrieval for IVF/ICSI. Semen donors and patients were recruited in accordance with local ethics approval (13/ES/0091) from the East of Scotland Research Ethics Service REC1. Activities of CatSper and KSper were assessed by patch clamp electrophysiology. Sperm [Ca2+]i responses were examined in sperm populations and single cells. Computer-assisted sperm analysis (CASA) parameters and penetration into viscous media were used to assess functional effects. hFF and progesterone significantly potentiated CatSper currents. Under quasi-physiological conditions, hFF (up to 50%) failed to alter membrane K+ conductance or current reversal potential. hFF and progesterone (at an equivalent concentration) stimulated similar biphasic [Ca2+]i signals both in sperm populations and single cells. At a high hFF concentration (10%), the sustained (plateau) component of the [Ca2+]i signal was consistently greater than that induced by progesterone alone. In single cell recordings, 1% hFF-induced [Ca2+]i oscillations similarly to progesterone but with 10% hFF generation of [Ca2+]i oscillations was suppressed. After treatment to ‘strip’ lipid-derived mediators, hFF failed to significantly stimulate CatSper currents but induced small [Ca2+]i responses that were greater than those induced by the equivalent concentration of progesterone after stripping. Similar [Ca2+]i responses were observed when sperm pretreated with 3 μM progesterone (to desensitize progesterone responses) were stimulated with hFF or stripped hFF. hFF stimulated viscous media penetration and was more effective than the equivalent does of progesterone. N/A. This was an in vitro study. Caution must be taken when extrapolating these results in vivo. This study directly demonstrates that hFF activates CatSper and establishes that the biologically important effects of hFF reflect, at least in part, action on this channel, primarily via progesterone. However, these experiments also demonstrate that other components of hFF both contribute to the [Ca2+]i signal and modulate the activation of CatSper. Simple in vitro experiments performed out of the context of the complex in vivo environment need to be interpreted with caution. Funding was provided by MRC (MR/K013343/1, MR/012492/1) (S.G.B., S.J.P., C.L.R.B.) and University of Abertay (sabbatical for S.G.B.). Additional funding was provided by TENOVUS SCOTLAND (S.M.D.S.), Chief Scientist Office/NHS Research Scotland (S.M.D.S). C.L.R.B. is EIC of MHR and Chair of the WHO ESG on Diagnosis of Male infertility. The remaining authors have no conlicts of interest.
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影响因子：
2.7
作者：
Bedu-Addo, K.;Barratt, C. L. R.;Publicover, S. J.
通讯作者：
Publicover, S. J.
DOI：
10.3791/1996
发表时间：
2010-06-16
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Nash, Katherine;Lefievre, Linda;Publicover, Stephen J
通讯作者：
Publicover, Stephen J
DOI：
10.1016/0002-9378(77)90690-1
发表时间：
1977-01-01
影响因子：
9.8
作者：
CHONG, AP;TAYMOR, ML;LECHENE, CP
通讯作者：
LECHENE, CP
DOI：
10.1016/j.bbrc.2017.01.010
发表时间：
2017-02-05
影响因子：
3.1
作者：
Lopez-Torres, Aidee S.;Gonzalez-Gonzalez, Maria E.;Chirinos, Mayel
通讯作者：
Chirinos, Mayel
影响因子：
13.3
作者：
Mortimer, David;Barratt, Christopher L. R.;Muller, Charles H.
通讯作者：
Muller, Charles H.