How Is Plasminogen/Plasmin System Contributing to Regulate Sperm Entry Into the Oocyte?

Grullón, Luis Alberto; Gadea, J.; Mondéjar, Irene; Matás, Carmen; Romar, Raquel; Coy, Pilar

doi:10.1177/1933719112473657

Cited by 20 publications

(15 citation statements)

References 32 publications

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“…This successful interaction were further confirmed through transmission electron (Additional file 2 : Figure S2) and confocal laser (Additional file 3 : Figure S3) microscopy analyses, showing higher accumulations of quantum dots in spermatozoa (TEM) that corresponded to similar area targeted by anti-plasminogen antibody (the sperm head). Overall and despite using different sources of antibodies (Acris and Santa Cruz), generated images appear alike, which position the use of nanoparticle-based imaging as a viable approach for non-invasive functional analyses of desired proteins, such as the plasminogen/plasmin system, having putative role during fertilization [ 22 , 24 ]. The current direct and indirect (through nanoparticles) findings bring further confirmation of the presence of plasminogen on mammalian oocytes [ 23 ], while revealing its presence in spermatozoa that has not yet been clearly demonstrated in the literature.…”

Section: Resultsmentioning

confidence: 99%

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Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

et al. 2015

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BackgroundThe fertility performance of animals is still a mystery and the full comprehension of mammalian gametes maturation and early embryonic development remains to be elucidated. The recent development in nanotechnology offers a new opportunity for real-time study of reproductive cells in their physiological environments. As a first step toward that goal, we evaluated the effectiveness of a fluorescent and luminescent nanoparticle for in vitro and ex vivo imaging of porcine gametes.MethodsFreshly harvested boar sperm were labeled with red-shifted (655 nm) quantum dot nanoparticles conjugated (QD+) or not (QD−) with plasminogen antibody and evaluated. Subsets of labeled spermatozoa were loaded into straws and placed within the lumen of gilt reproductive tracts for ex vivo intra-uterine imaging. Porcine cumulus-oocyte complexes (COCs) were matured in the presence of QD− or QD+. Ovarian follicles were microinjected with QD− or QD+ and placed in culture for up to 4 days. After labeling, all samples were supplemented with coelenterazine, the luciferase substrate, and immediately submitted to bioluminescence analysis, followed by fluorescence and hyperspectral imaging. Data were analyzed with ANOVA and P < 0.05 indicated significant differences.ResultsAll labeled-samples revealed bioluminescence emission that was confirmed by fluorescence and hyperspectral imaging of the QD localization within the cells and tissues. Over 76% of spermatozoa and both immature and mature COCs were successfully labeled with QD− or QD+. The QD− fluorescence appeared homogenously distributed in the oocytes, while found in the entire sperm length with a higher accumulation within the mid-piece. Labeled-follicles exhibited a progressive migration of QD nanoparticles within the follicle wall during culture. In contrast, QD+ fluorescence signals appeared condensed and stronger in the follicle cells, sperm head, and sub-plasma membrane area of mature oocytes. Weaker QD+ signals were detected in the cumulus cells. Fluorescence and hyperspectral microscope imaging showed comparable intracellular QD localization. Ex-vivo intra-uterine bioluminescence imaging of labeled spermatozoa revealed stronger signals captured over the oviducts, with uterine body allowing the lowest signal detection.ConclusionFindings indicate that conjugated and non-conjugated fluorescent nanoparticles can be used for effective labeling of mammalian gametes for in vitro monitoring and potential in vivo targeted-imaging.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-015-0097-1) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

“…In parallel, we conducted a targeted bio-imaging using the QD-BRET tagged with anti-porcine plasminogen antibody. In previous studies, this protein has been detected in the oocyte and has putative role during fertilization [ 22 – 24 ].…”

Section: Introductionmentioning

confidence: 99%

Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

et al. 2015

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“…It is of note that this protein, which converts plasminogen into the serine protease plasmin, was only detected in ODF samples collected from the ampullary segment. The fibrinolytic plasmin/plasminogen system affects gamete interaction . Together with the matrix metalloproteinase (MMP) system, this protease system also degrades and remodels the extracellular matrix (ECM) .…”

Section: Discussionmentioning

confidence: 99%

Identification of Rabbit Oviductal Fluid Proteins Involved in Pre‐Fertilization Processes by Quantitative Proteomics

Hackenbroch

Meyer

et al. 2019

Proteomics

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Oviductal fluid (ODF) proteins modulate and support reproductive processes in the oviduct. In the present study, proteins involved in the biological events that precede fertilization have been identified in the rabbit ODF proteome, isolated from the ampulla and isthmus of the oviduct at different time points within 8 h after intrauterine insemination. A workflow is used that integrates lectin affinity capture with stable‐isotope dimethyl labeling prior to nanoLC‐MS/MS analysis. In total, over 400 ODF proteins, including 214 lectin enriched glycoproteins, are identified and quantified. Selected data are validated by Western blot analysis. Spatiotemporal alterations in the abundance of ODF proteins in response to insemination are detected by global analysis. A subset of 63 potentially biologically relevant ODF proteins is identified, including extracellular matrix components, chaperones, oxidoreductases, and immunity proteins. Functional enrichment analysis reveals an altered peptidase regulator activity upon insemination. In addition to protein identification and abundance changes, N‐glycopeptide analysis further identifies 281 glycosites on 199 proteins. Taken together, these results show, for the first time, the evolving oviductal milieu early upon insemination. The identified proteins are likely those that modulate in vitro processes, including spermatozoa function.

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Section: Introductionmentioning

confidence: 83%

Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

Feugang¹,

Youngblood²,

Greene³

et al. 2016

Nano Online

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Background: The fertility performance of animals is still a mystery and the full comprehension of mammalian gametes maturation and early embryonic development remains to be elucidated. The recent development in nanotechnology offers a new opportunity for real-time study of reproductive cells in their physiological environments. As a first step toward that goal, we evaluated the effectiveness of a fluorescent and luminescent nanoparticle for in vitro and ex vivo imaging of porcine gametes. Methods:Freshly harvested boar sperm were labeled with red-shifted (655 nm) quantum dot nanoparticles conjugated (QD+) or not (QD−) with plasminogen antibody and evaluated. Subsets of labeled spermatozoa were loaded into straws and placed within the lumen of gilt reproductive tracts for ex vivo intra-uterine imaging. Porcine cumulusoocyte complexes (COCs) were matured in the presence of QD− or QD+. Ovarian follicles were microinjected with QD− or QD+ and placed in culture for up to 4 days. After labeling, all samples were supplemented with coelenterazine, the luciferase substrate, and immediately submitted to bioluminescence analysis, followed by fluorescence and hyperspectral imaging. Data were analyzed with ANOVA and P < 0.05 indicated significant differences. Results:All labeled-samples revealed bioluminescence emission that was confirmed by fluorescence and hyperspectral imaging of the QD localization within the cells and tissues. Over 76% of spermatozoa and both immature and mature COCs were successfully labeled with QD− or QD+. The QD− fluorescence appeared homogenously distributed in the oocytes, while found in the entire sperm length with a higher accumulation within the mid-piece. Labeled-follicles exhibited a progressive migration of QD nanoparticles within the follicle wall during culture. In contrast, QD+ fluorescence signals appeared condensed and stronger in the follicle cells, sperm head, and sub-plasma membrane area of mature oocytes. Weaker QD+ signals were detected in the cumulus cells. Fluorescence and hyperspectral microscope imaging showed comparable intracellular QD localization. Ex-vivo intra-uterine bioluminescence imaging of labeled spermatozoa revealed stronger signals captured over the oviducts, with uterine body allowing the lowest signal detection. Conclusion: Findings indicate that conjugated and non-conjugated fluorescent nanoparticles can be used for effective labeling of mammalian gametes for in vitro monitoring and potential in vivo targeted-imaging.

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How Is Plasminogen/Plasmin System Contributing to Regulate Sperm Entry Into the Oocyte?

Cited by 20 publications

References 32 publications

Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

Identification of Rabbit Oviductal Fluid Proteins Involved in Pre‐Fertilization Processes by Quantitative Proteomics

Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

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