Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots
To date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study in...
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| Format: | Article |
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MDPI AG
2023-06-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/12/13/2397 |
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| author | Giulia De Angelis Camilla Badiali Laura Chronopoulou Cleofe Palocci Gabriella Pasqua |
| author_facet | Giulia De Angelis Camilla Badiali Laura Chronopoulou Cleofe Palocci Gabriella Pasqua |
| author_sort | Giulia De Angelis |
| collection | DOAJ |
| description | To date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study investigated its role in the uptake of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs). The experiments were performed on cultured cells and roots of <i>Arabidopsis thaliana</i>. Dynasore was used to inhibit the activity of dynamin-like proteins to investigate whether PLGA NPs enter plant cells through a dynamin-like-dependent or dynamin-like-independent endocytic pathway. Observations were performed by confocal microscopy using a fluorescent probe, coumarin 6, loaded in PLGA NPs. The results showed that both cells and roots of <i>A. thaliana</i> rapidly take up PLGA NPs. Dynasore was administered at different concentrations and exposure times in order to identify the effective ones for inhibitory activity. Treatments with dynasore did not prevent the NPs uptake, as revealed by the presence of fluorescence emission detected in the cytoplasm. At the highest concentration and the longest exposure time to dynasore, the fluorescence of NPs was not visible due to cell death. Thus, the results suggest that, because the NPs’ uptake is unaffected by dynasore exposure, NPs can enter cells and roots by following a dynamin-like-independent endocytic pathway. |
| first_indexed | 2024-03-11T01:31:57Z |
| format | Article |
| id | doaj.art-1495d9e66f864b6280418ec4271e6b06 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-11T01:31:57Z |
| publishDate | 2023-06-01 |
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| series | Plants |
| spelling | doaj.art-1495d9e66f864b6280418ec4271e6b062023-11-18T17:16:50ZengMDPI AGPlants2223-77472023-06-011213239710.3390/plants12132397Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet RootsGiulia De Angelis0Camilla Badiali1Laura Chronopoulou2Cleofe Palocci3Gabriella Pasqua4Department of Environmental Biology, Sapienza University of Rome, P. le Aldo Moro 5, 00185 Rome, ItalyDepartment of Environmental Biology, Sapienza University of Rome, P. le Aldo Moro 5, 00185 Rome, ItalyDepartment of Chemistry, Sapienza University of Rome, P. le Aldo Moro 5, 00185 Rome, ItalyDepartment of Chemistry, Sapienza University of Rome, P. le Aldo Moro 5, 00185 Rome, ItalyDepartment of Environmental Biology, Sapienza University of Rome, P. le Aldo Moro 5, 00185 Rome, ItalyTo date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study investigated its role in the uptake of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs). The experiments were performed on cultured cells and roots of <i>Arabidopsis thaliana</i>. Dynasore was used to inhibit the activity of dynamin-like proteins to investigate whether PLGA NPs enter plant cells through a dynamin-like-dependent or dynamin-like-independent endocytic pathway. Observations were performed by confocal microscopy using a fluorescent probe, coumarin 6, loaded in PLGA NPs. The results showed that both cells and roots of <i>A. thaliana</i> rapidly take up PLGA NPs. Dynasore was administered at different concentrations and exposure times in order to identify the effective ones for inhibitory activity. Treatments with dynasore did not prevent the NPs uptake, as revealed by the presence of fluorescence emission detected in the cytoplasm. At the highest concentration and the longest exposure time to dynasore, the fluorescence of NPs was not visible due to cell death. Thus, the results suggest that, because the NPs’ uptake is unaffected by dynasore exposure, NPs can enter cells and roots by following a dynamin-like-independent endocytic pathway.https://www.mdpi.com/2223-7747/12/13/2397endocytosisdynamin-independent endocytosisPLGA nanoparticlesdynasorecell culturesroot plantlets |
| spellingShingle | Giulia De Angelis Camilla Badiali Laura Chronopoulou Cleofe Palocci Gabriella Pasqua Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots Plants endocytosis dynamin-independent endocytosis PLGA nanoparticles dynasore cell cultures root plantlets |
| title | Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots |
| title_full | Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots |
| title_fullStr | Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots |
| title_full_unstemmed | Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots |
| title_short | Confocal Microscopy Investigations of Biopolymeric PLGA Nanoparticle Uptake in <i>Arabidopsis thaliana</i> L. Cultured Cells and Plantlet Roots |
| title_sort | confocal microscopy investigations of biopolymeric plga nanoparticle uptake in i arabidopsis thaliana i l cultured cells and plantlet roots |
| topic | endocytosis dynamin-independent endocytosis PLGA nanoparticles dynasore cell cultures root plantlets |
| url | https://www.mdpi.com/2223-7747/12/13/2397 |
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