Caveolae-mediated Endocytosis of Intratracheally Instilled Gold Colloid Nanoparticles at the Air–Blood Barrier in Mice

Naota, Misaki; Shimada, Akinori; Morita, Takehito; Yamamoto, Yuko; Inoue, Ken‐ichiro; Takano, Hirohisa

doi:10.1177/0192623312457271

Cited by 14 publications

(12 citation statements)

References 28 publications

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“…The uptake of Au NPs via caveolinmediated endocytosis has already been observed in previous studies [23,51,57,58]. These studies, however, used different models and differently modified and sized particles.…”

Section: Cellular Np Uptakementioning

confidence: 52%

“…For example, Hao and colleagues found a m cd-mediated uptake inhibition of 10 nm cysteincy5-coated Au NPs in HeLa cells [51]. The results of an in vivo study by Naota and colleagues showed localization of 20-nm plain, colloidal Au NPs in caveolin-1-positive stained vesicles in air-blood barrier cells in mice [58]. In a previous study with A549 cells and 15-nm sized plain and polyethylene glycol-coated Au NPs we found a m cd-mediated inhibition after 1 h of 86 and 97%, respectively [33].…”

Section: Cellular Np Uptakementioning

confidence: 99%

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Quantification of Gold Nanoparticle Cell Uptake Under Controlled Biological Conditions and Adequate Resolution

et al. 2014

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Aim:We examined cellular uptake mechanisms of fluorescently labeled polymer-coated gold nanoparticles (NPs) under different biological conditions by two quantitative, microscopic approaches. Materials & methods: Uptake mechanisms were evaluated using endocytotic inhibitors that were tested for specificity and cytotoxicity. Cellular uptake of gold NPs was analyzed either by laser scanning microscopy or transmission electron microscopy, and quantified by means of stereology using cells from the same experiment. Results: Optimal inhibitor conditions were only achieved with chlorpromazine (clathrin-mediated endocytosis) and methyl--cyclodextrin (caveolinmediated endocytosis). A significant methyl--cyclodextrin-mediated inhibition (63-69%) and chlorpromazine-mediated increase (43-98%) of intracellular NPs was demonstrated with both imaging techniques, suggesting a predominant uptake via caveolin-medicated endocytois. Transmission electron microscopy imaging revealed more than 95% of NPs localized in intracellular vesicles and approximately 150-times more NP events/cell were detected than by laser scanning microscopy. Conclusion: We emphasize the importance of studying NP-cell interactions under controlled experimental conditions and at adequate microscopic resolution in combination with stereology. Keywords:In recent years, various types of nanoparticles (NPs; <100 nm in all three dimensions, ISO TS 27687:2008) have been designed for potential biomedical and pharmaceutical applications, such as vehicles for targeted drug delivery, nucleic acids, biomedical imaging or biosensing [1][2][3]. Therefore, an increasing number of research studies have focused on the interaction of NPs with biocellular systems [4,5]. To analyze different types of NP-cell interactions, such as cellular NP uptake, it is essential to have valuable techniques for visualization and quantification, which are mainly based on spectroscopy or microscopy [6].The method of choice for the intracellular detection of NPs depends on the characteristics of the particles (fluorescence, size, and structure or electron density) and on the cellular structure of interest. Advantages of spectroscopy and different microscopic techniques have been reviewed in detail elsewhere [6,7]. In brief, spectroscopy such as inductively coupled plasma-mass spectroscopy [8] allows the quantification and detection of chemical NP elements within the cell, but does not provide any further information on the NP location within the cell. Fluorescence or laser scanning microscopy (LSM) allows real-time analysis or co-localization studies with fluorescently labeled structures [9]. Transmission electron microscopy (TEM) provides the highest resolution and allows quantification of NPs in sub cellular structures [10].

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Section: Cellular Np Uptakementioning

confidence: 52%

Section: Cellular Np Uptakementioning

confidence: 99%

Quantification of Gold Nanoparticle Cell Uptake Under Controlled Biological Conditions and Adequate Resolution

et al. 2014

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“…Caveolae-mediated endocytosis is also considered to be one of the pathways for the translocation of nanoparticles in the air–blood barrier [26]. Caveolin-1 protein plays a role in the regulation of caveolar invagination and the formation of caveolae, which is one form of the endocytic vesicle [19, 25, 27, 28].…”

Section: Discussionmentioning

confidence: 99%

Demonstration of the Clathrin- and Caveolin-Mediated Endocytosis at the Maternal–Fetal Barrier in Mouse Placenta after Intravenous Administration of Gold Nanoparticles

et al. 2014

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Exposure to nanoparticles during pregnancy is a public concern, because nanoparticles may pass from the mother to the fetus across the placenta. The purpose of this study was to determine the possible translocation pathway of gold nanoparticles across the maternal–fetal barrier as well as the toxicity of intravenously administered gold nanoparticles to the placenta and fetus. Pregnant ICR mice were intravenously injected with 0.01% of 20- and 50-nm gold nanoparticle solutions on the 16th and 17th days of gestation. There was no sign of toxic damage to the placentas as well as maternal and fetal organs of the mice treated with 20- and 50-nm gold nanoparticles. ICP-MS analysis demonstrated significant amounts of gold deposited in the maternal livers and placentas, but no detectable level of gold in the fetal organs. However, electron microscopy demonstrated an increase of endocytic vesicles in the cytoplasm of syncytiotrophoblasts and fetal endothelial cells in the maternal–fetal barrier of mice treated with gold nanoparticles. Clathrin immunohistochemistry and immunoblotting showed increased immunoreactivity of clathrin protein in the placental tissues of mice treated with 20- and 50-nm gold nanoparticles; clathrin immunopositivity was observed in syncytiotrophoblasts and fetal endothelial cells. In contrast, caveolin-1 immunopositivity was observed exclusively in the fetal endothelium. These findings suggested that intravenous administration of gold nanoparticles may upregulate clathrin- and caveolin-mediated endocytosis at the maternal–fetal barrier in mouse placenta.

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“…Within AJ zona occludins establish links to the tubulin-based cytoskeleton, as well as actin; Cav-1 appears to be necessary for retrieval of these junctional proteins[122]. Caveolar-type MLR are also involved in endocytosis by epithelial cells[123], which in some instances involves Rab11 and the tubulin-based cytoskeleton[124]. Additional findings indicate that basolateral membranes may contain different Cavs[125, 126] and lipid components[127, 128] than those of the apical membranes of epithelial cells.…”

Section: Introductionmentioning

confidence: 99%

Interaction of membrane/lipid rafts with the cytoskeleton: Impact on signaling and function

Head

Patel

Insel

2014

Biochimica et Biophysica Acta (BBA) - Biomembranes

435

192

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Summary The plasma membrane in eukaryotic cells contains microdomains that are enriched in certain glycosphingolipids, gangliosides, and sterols (such as cholesterol) to form membrane/lipid rafts (MLR). These regions exist as caveolae, morphologically observable flask-like invaginations, or as a less easily detectable planar form. MLR are scaffolds for many molecular entities, including signaling receptors and ion channels that communicate extracellular stimuli to the intracellular milieu. Much evidence indicates that this organization and/or the clustering of MLR into more active signaling platforms depends upon interactions with and dynamic rearrangement of the cytoskeleton. Several cytoskeletal components and binding partners, as well as enzymes that regulate the cytoskeleton, localize to MLR and help regulate lateral diffusion of membrane proteins and lipids in response to extracellular events (e.g., receptor activation, shear stress, electrical conductance, and nutrient demand). MLR regulate cellular polarity, adherence to the extracellular matrix, signaling events (including ones that affect growth and migration), and are sites of cellular entry of certain pathogens, toxins and nanoparticles. The dynamic interaction between MLR and the underlying cytoskeleton thus regulates many facets of the function of eukaryotic cells and their adaptation to changing environments. Here, we review general features of MLR and caveolae and their role in several aspects of cellular function, including polarity of endothelial and epithelial cells, cell migration, mechanotransduction, lymphocyte activation, neuronal growth and signaling, and a variety of disease settings.

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Caveolae-mediated Endocytosis of Intratracheally Instilled Gold Colloid Nanoparticles at the Air–Blood Barrier in Mice

Cited by 14 publications

References 28 publications

Quantification of Gold Nanoparticle Cell Uptake Under Controlled Biological Conditions and Adequate Resolution

Quantification of Gold Nanoparticle Cell Uptake Under Controlled Biological Conditions and Adequate Resolution

Demonstration of the Clathrin- and Caveolin-Mediated Endocytosis at the Maternal–Fetal Barrier in Mouse Placenta after Intravenous Administration of Gold Nanoparticles

Interaction of membrane/lipid rafts with the cytoskeleton: Impact on signaling and function

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