Application of quantum dot nanoparticles for potential non-invasive bio-imaging of mammalian spermatozoa.
Application of quantum dot nanoparticles for potential non-invasive bio-imaging of mammalian spermatozoa.
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DOI：
10.1186/1477-3155-10-45
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发表时间：
2012-12-14
影响因子：
10.2
通讯作者：
Ryan PL
中科院分区：
文献类型：
--
作者：
Feugang JM;Youngblood RC;Greene JM;Fahad AS;Monroe WA;Willard ST;Ryan PL
哺乳动物精子在通过女性生殖道的过程中会遇到各种障碍，只有很少或根本没有到达受精部位。目前，用于非侵入性研究授精后精子迁移的技术方法有限。由于有关整个女性生殖道精子行为的知识仍然难以捉摸，最近开发的自发光量子点纳米粒子可能为实时体外和体内精子监测提供潜在手段。在这里，我们展示了公猪精子无害地相互作用并结合生物发光共振能量转移共轭量子点（BRET-QD）纳米颗粒的能力。共聚焦显微镜显示了整个精子中 BRET-QD 的原位荧光，而使用透射电子显微镜的超微结构分析表明 BRET-QD 定位于精子质膜和细胞内区室。在体外受控测定中，生物发光成像表明，与 BRET-QD 和荧光素酶底物（腔肠素）一起孵育的精子发出高于背景的光（光子/秒），这证实了原位荧光成像。最重要的是，当以适当的比例使用时，精子活力、活力和受精潜力不会受到 BRET-QD 掺入的影响。我们的结果表明，在适当的平衡下使用时，猪精子可以掺入 BRET-QD 纳米颗粒，而不影响其运动性和与卵母细胞相互作用的能力。我们预计我们的研究将能够利用这种新方法在分子水平上深入探索体内迁移、受精和胚胎发育的男性组成部分。
Various obstacles are encountered by mammalian spermatozoa during their journey through the female genital tract, and only few or none will reach the site of fertilization. Currently, there are limited technical approaches for non-invasive investigation of spermatozoa migration after insemination. As the knowledge surrounding sperm behavior throughout the female genital tract still remains elusive, the recent development of self-illuminating quantum dot nanoparticles may present a potential means for real-time in vitro and in vivo monitoring of spermatozoa. Here, we show the ability of boar spermatozoa to harmlessly interact and incorporate bioluminescent resonance energy transfer-conjugated quantum dot (BRET-QD) nanoparticles. The confocal microscope revealed in situ fluorescence of BRET-QD in the entire spermatozoon, while the ultra-structural analysis using the transmission electron microscope indicated BRET-QD localization on the sperm plasma membrane and intracellular compartment. In controlled-in vitro assays, bioluminescent imaging demonstrated that spermatozoa incubated with BRET-QD and luciferase substrate (coelenterazine) emit light (photons/sec) above the background, which confirmed the in situ fluorescence imaging. Most importantly, sperm motility, viability, and fertilizing potential were not affected by the BRET-QD incorporation when used at an appropriated ratio. Our results demonstrate that pig spermatozoa can incorporate BRET-QD nanoparticles without affecting their motility and capacity to interact with the oocyte when used at an appropriated balance. We anticipate that our study will enable in-depth exploration of the male components of in vivo migration, fertilization, and embryonic development at the molecular level using this novel approach.
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