Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching
<p>Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
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Society of Photo-optical Instrumentation Engineers (SPIE)
2014
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| _version_ | 1797050329377800192 |
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| author | Kuzma-Kuzniarska, M Yapp, C Pearson-Jones, T Jones, A Hulley, P |
| author_facet | Kuzma-Kuzniarska, M Yapp, C Pearson-Jones, T Jones, A Hulley, P |
| author_sort | Kuzma-Kuzniarska, M |
| collection | OXFORD |
| description | <p>Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to investigate gap junction-mediated intercellular communication in healthy human tendon-derived cells using fluorescence recovery after photobleaching (FRAP). The FRAP is a noninvasive technique that allows quantitative measurement of gap junction function in living cells. It is based on diffusion-dependent redistribution of a gap junction-permeable fluorescent dye. Using FRAP, we showed that human tenocytes form functional gap junctions in monolayer and three-dimensional (3-D) collagen I culture. Fluorescently labeled tenocytes following photobleaching rapidly reacquired the fluorescent dye from neighboring cells, while HeLa cells, which do not communicate by gap junctions, remained bleached. Furthermore, both <b>18</b> β-glycyrrhetinic acid and carbenoxolone, standard inhibitors of gap junction activity, impaired fluorescence recovery in tendon cells. In both monolayer and 3-D cultures, intercellular communication in isolated cells was significantly decreased when compared with cells forming many cell-to-cell contacts. In this study, we used FRAP as a tool to quantify and experimentally manipulate the function of gap junctions in human tenocytes in both two-dimensional (2-D) and 3-D cultures.</p> |
| first_indexed | 2024-03-06T18:03:34Z |
| format | Journal article |
| id | oxford-uuid:00a46418-fb47-4b63-bd04-013b4284eb3e |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T18:03:34Z |
| publishDate | 2014 |
| publisher | Society of Photo-optical Instrumentation Engineers (SPIE) |
| record_format | dspace |
| spelling | oxford-uuid:00a46418-fb47-4b63-bd04-013b4284eb3e2022-03-26T08:30:39ZFunctional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleachingJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:00a46418-fb47-4b63-bd04-013b4284eb3eEnglishSymplectic Elements at OxfordSociety of Photo-optical Instrumentation Engineers (SPIE)2014Kuzma-Kuzniarska, MYapp, CPearson-Jones, TJones, AHulley, P <p>Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to investigate gap junction-mediated intercellular communication in healthy human tendon-derived cells using fluorescence recovery after photobleaching (FRAP). The FRAP is a noninvasive technique that allows quantitative measurement of gap junction function in living cells. It is based on diffusion-dependent redistribution of a gap junction-permeable fluorescent dye. Using FRAP, we showed that human tenocytes form functional gap junctions in monolayer and three-dimensional (3-D) collagen I culture. Fluorescently labeled tenocytes following photobleaching rapidly reacquired the fluorescent dye from neighboring cells, while HeLa cells, which do not communicate by gap junctions, remained bleached. Furthermore, both <b>18</b> β-glycyrrhetinic acid and carbenoxolone, standard inhibitors of gap junction activity, impaired fluorescence recovery in tendon cells. In both monolayer and 3-D cultures, intercellular communication in isolated cells was significantly decreased when compared with cells forming many cell-to-cell contacts. In this study, we used FRAP as a tool to quantify and experimentally manipulate the function of gap junctions in human tenocytes in both two-dimensional (2-D) and 3-D cultures.</p> |
| spellingShingle | Kuzma-Kuzniarska, M Yapp, C Pearson-Jones, T Jones, A Hulley, P Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
| title | Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
| title_full | Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
| title_fullStr | Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
| title_full_unstemmed | Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
| title_short | Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
| title_sort | functional assessment of gap junctions in monolayer and three dimensional cultures of human tendon cells using fluorescence recovery after photobleaching |
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