Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation
Neurodevelopmental and neurodegenerative disorders are characterized by subtle alterations in synaptic connections and perturbed neuronal network functionality. A hallmark of neuronal connectivity is the presence of dendritic spines, micron-sized protrusions of the dendritic shaft that compartmental...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-03-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fncel.2018.00080/full |
| _version_ | 1819180438555459584 |
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| author | Ranhua Xiong Ranhua Xiong Peter Verstraelen Jo Demeester Andre G. Skirtach Andre G. Skirtach Jean-Pierre Timmermans Stefaan C. De Smedt Stefaan C. De Smedt Winnok H. De Vos Winnok H. De Vos Winnok H. De Vos Kevin Braeckmans Kevin Braeckmans Kevin Braeckmans Kevin Braeckmans |
| author_facet | Ranhua Xiong Ranhua Xiong Peter Verstraelen Jo Demeester Andre G. Skirtach Andre G. Skirtach Jean-Pierre Timmermans Stefaan C. De Smedt Stefaan C. De Smedt Winnok H. De Vos Winnok H. De Vos Winnok H. De Vos Kevin Braeckmans Kevin Braeckmans Kevin Braeckmans Kevin Braeckmans |
| author_sort | Ranhua Xiong |
| collection | DOAJ |
| description | Neurodevelopmental and neurodegenerative disorders are characterized by subtle alterations in synaptic connections and perturbed neuronal network functionality. A hallmark of neuronal connectivity is the presence of dendritic spines, micron-sized protrusions of the dendritic shaft that compartmentalize single synapses to fine-tune synaptic strength. However, accurate quantification of spine density and morphology in mature neuronal networks is hampered by the lack of targeted labeling strategies. To resolve this, we have optimized a method to deliver cell-impermeable compounds into selected cells based on Spatially resolved NAnoparticle-enhanced Photoporation (SNAP). We show that SNAP enables efficient labeling of selected individual neurons and their spines in dense cultured networks without affecting short-term viability. We compare SNAP with widely used spine labeling techniques such as the application of lipophilic dyes and genetically encoded fluorescent markers. Using SNAP, we demonstrate a time-dependent increase in spine density in healthy cultures as well as a reduction in spine density after chemical mimicry of hypoxia. Since the sparse labeling procedure can be automated using an intelligent acquisition scheme, SNAP holds promise for high-content screening campaigns of neuronal connectivity in the context of neurodevelopmental and neurodegenerative disorders. |
| first_indexed | 2024-12-22T22:14:21Z |
| format | Article |
| id | doaj.art-a4e68f9c39c843f9a3325154ee168073 |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-12-22T22:14:21Z |
| publishDate | 2018-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-a4e68f9c39c843f9a3325154ee1680732022-12-21T18:10:49ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022018-03-011210.3389/fncel.2018.00080329232Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced PhotoporationRanhua Xiong0Ranhua Xiong1Peter Verstraelen2Jo Demeester3Andre G. Skirtach4Andre G. Skirtach5Jean-Pierre Timmermans6Stefaan C. De Smedt7Stefaan C. De Smedt8Winnok H. De Vos9Winnok H. De Vos10Winnok H. De Vos11Kevin Braeckmans12Kevin Braeckmans13Kevin Braeckmans14Kevin Braeckmans15Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumCentre for Nano- and Biophotonics, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumLaboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, Antwerp, BelgiumLaboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumCentre for Nano- and Biophotonics, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumDepartment of Molecular Biotechnology, Ghent University, Ghent, BelgiumLaboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, Antwerp, BelgiumLaboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumCollege of Chemical Engineering, Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, Nanjing Forestry University, Nanjing, ChinaCentre for Nano- and Biophotonics, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumLaboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, Antwerp, BelgiumDepartment of Molecular Biotechnology, Ghent University, Ghent, BelgiumLaboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumCentre for Nano- and Biophotonics, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, BelgiumUniv Lille 1, Univ Lille Nord France, IEMN, UMR 8520, Villeneuve D’Ascq, FranceUniv Lille 1, Univ Lille Nord France, Lab Phys Lasers Atomes & Mol, UMR 8523, Villeneuve D’Ascq, FranceNeurodevelopmental and neurodegenerative disorders are characterized by subtle alterations in synaptic connections and perturbed neuronal network functionality. A hallmark of neuronal connectivity is the presence of dendritic spines, micron-sized protrusions of the dendritic shaft that compartmentalize single synapses to fine-tune synaptic strength. However, accurate quantification of spine density and morphology in mature neuronal networks is hampered by the lack of targeted labeling strategies. To resolve this, we have optimized a method to deliver cell-impermeable compounds into selected cells based on Spatially resolved NAnoparticle-enhanced Photoporation (SNAP). We show that SNAP enables efficient labeling of selected individual neurons and their spines in dense cultured networks without affecting short-term viability. We compare SNAP with widely used spine labeling techniques such as the application of lipophilic dyes and genetically encoded fluorescent markers. Using SNAP, we demonstrate a time-dependent increase in spine density in healthy cultures as well as a reduction in spine density after chemical mimicry of hypoxia. Since the sparse labeling procedure can be automated using an intelligent acquisition scheme, SNAP holds promise for high-content screening campaigns of neuronal connectivity in the context of neurodevelopmental and neurodegenerative disorders.http://journal.frontiersin.org/article/10.3389/fncel.2018.00080/fulldendritic spinegold nanoparticlephotoporationprimary neuronal cultureneuron labelingSNAP |
| spellingShingle | Ranhua Xiong Ranhua Xiong Peter Verstraelen Jo Demeester Andre G. Skirtach Andre G. Skirtach Jean-Pierre Timmermans Stefaan C. De Smedt Stefaan C. De Smedt Winnok H. De Vos Winnok H. De Vos Winnok H. De Vos Kevin Braeckmans Kevin Braeckmans Kevin Braeckmans Kevin Braeckmans Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation Frontiers in Cellular Neuroscience dendritic spine gold nanoparticle photoporation primary neuronal culture neuron labeling SNAP |
| title | Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation |
| title_full | Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation |
| title_fullStr | Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation |
| title_full_unstemmed | Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation |
| title_short | Selective Labeling of Individual Neurons in Dense Cultured Networks With Nanoparticle-Enhanced Photoporation |
| title_sort | selective labeling of individual neurons in dense cultured networks with nanoparticle enhanced photoporation |
| topic | dendritic spine gold nanoparticle photoporation primary neuronal culture neuron labeling SNAP |
| url | http://journal.frontiersin.org/article/10.3389/fncel.2018.00080/full |
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