Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3)
Connexin (Cx) proteins form gap junction channels (GJC) and hemichannels that a allow bidirectional flow of ions and metabolites between the cytoplasm and extracellular space, respectively. Under physiological conditions, hemichannels have a very low probability of opening, but in certain pathologie...
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Frontiers Media S.A.
2020-01-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2019.01574/full |
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| author | Vania A. Figueroa Vania A. Figueroa Oscar Jara Carolina A. Oliva Marcelo Ezquer Fernando Ezquer Mauricio A. Retamal Mauricio A. Retamal Mauricio A. Retamal Agustín D. Martínez Guillermo A. Altenberg Guillermo A. Altenberg Aníbal A. Vargas Aníbal A. Vargas |
| author_facet | Vania A. Figueroa Vania A. Figueroa Oscar Jara Carolina A. Oliva Marcelo Ezquer Fernando Ezquer Mauricio A. Retamal Mauricio A. Retamal Mauricio A. Retamal Agustín D. Martínez Guillermo A. Altenberg Guillermo A. Altenberg Aníbal A. Vargas Aníbal A. Vargas |
| author_sort | Vania A. Figueroa |
| collection | DOAJ |
| description | Connexin (Cx) proteins form gap junction channels (GJC) and hemichannels that a allow bidirectional flow of ions and metabolites between the cytoplasm and extracellular space, respectively. Under physiological conditions, hemichannels have a very low probability of opening, but in certain pathologies, hemichannels activity can increase and induce and/or accelerate cell death. Several mechanisms control hemichannels activity, including phosphorylation and oxidation (i.e., S-nitrosylation). Recently, the effect of polyunsaturated fatty acids (PUFAs) such as linoleic acid (LA), were found to modulate Cxs. It has been seen that LA increase cell death in bovine and human lens cells. The lens is a structure allocated in the eye that highly depends on Cx for the metabolic coupling between its cells, a condition necessary for its transparency. Therefore, we hypothesized that LA induces lens cells death by modulating hemichannel activity. In this work, we characterized the effect of LA on hemichannel activity and survival of HLE-B3 cells (a human lens epithelial cell line). We found that HLE-B3 cells expresses Cx43, Cx46, and Cx50 and can form functional hemichannels in their plasma membrane. The extracellular exposure to 10–50 μM of LA increases hemichannels activity (dye uptake) in a concentration-dependent manner, which was reduced by Cx-channel blockers, such as the Cx-mimetic peptide Gap27 and TATGap19, La3+, carbenoxolone (CBX) and the Akt kinase inhibitor. Additionally, LA increases intracellular calcium, which is attenuated in the presence of TATGap19, a specific Cx43-hemichannel inhibitor. Finally, the long exposure of HLE-B3 cells to LA 20 and 50 μM, reduced cell viability, which was prevented by CBX. Moreover, LA increased the proportion of apoptotic HLE-B3 cells, effect that was prevented by the Cx-mimetic peptide TAT-Gap19 but not by Akt inhibitor. Altogether, these findings strongly suggest a contribution of hemichannels opening in the cell death induced by LA in HLE-B3 cells. These cells can be an excellent tool to develop pharmacological studies in vitro. |
| first_indexed | 2024-12-12T02:05:44Z |
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| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-12T02:05:44Z |
| publishDate | 2020-01-01 |
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| series | Frontiers in Physiology |
| spelling | doaj.art-fc85ee9db0084b9fbe39cd93fda06e802022-12-22T00:42:04ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2020-01-011010.3389/fphys.2019.01574447590Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3)Vania A. Figueroa0Vania A. Figueroa1Oscar Jara2Carolina A. Oliva3Marcelo Ezquer4Fernando Ezquer5Mauricio A. Retamal6Mauricio A. Retamal7Mauricio A. Retamal8Agustín D. Martínez9Guillermo A. Altenberg10Guillermo A. Altenberg11Aníbal A. Vargas12Aníbal A. Vargas13Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, Santiago, ChileInstituto de Ciencias de la Salud, Universidad de O’Higgins, Rancagua, ChileDepartment of Pediatrics, University of Chicago, Chicago, IL, United StatesCentro de Envejecimiento y Regeneración (CARE-UC), Departamento Biología Celular y Molecular, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, ChileCentro de Medicina Regenerativa, Facultad de Medicina, Clínica Alemana Universidad del Desarrollo, Santiago, ChileCentro de Medicina Regenerativa, Facultad de Medicina, Clínica Alemana Universidad del Desarrollo, Santiago, ChileUniversidad del Desarrollo, Centro de Fisiología Celular e Integrativa, Facultad de Medicina Clínica Alemana, Santiago, ChileDepartment of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center, Lubbock, TX, United StatesCenter for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, TX, United StatesCentro Interdisciplinario de Neurociencia de Valparaíso, Instituto de Neurociencia, Facultad de Ciencias, Universidad de Valparaíso, Valparaíso, ChileDepartment of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center, Lubbock, TX, United StatesCenter for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, TX, United StatesInstituto de Ciencias de la Salud, Universidad de O’Higgins, Rancagua, ChileCentro Interdisciplinario de Neurociencia de Valparaíso, Instituto de Neurociencia, Facultad de Ciencias, Universidad de Valparaíso, Valparaíso, ChileConnexin (Cx) proteins form gap junction channels (GJC) and hemichannels that a allow bidirectional flow of ions and metabolites between the cytoplasm and extracellular space, respectively. Under physiological conditions, hemichannels have a very low probability of opening, but in certain pathologies, hemichannels activity can increase and induce and/or accelerate cell death. Several mechanisms control hemichannels activity, including phosphorylation and oxidation (i.e., S-nitrosylation). Recently, the effect of polyunsaturated fatty acids (PUFAs) such as linoleic acid (LA), were found to modulate Cxs. It has been seen that LA increase cell death in bovine and human lens cells. The lens is a structure allocated in the eye that highly depends on Cx for the metabolic coupling between its cells, a condition necessary for its transparency. Therefore, we hypothesized that LA induces lens cells death by modulating hemichannel activity. In this work, we characterized the effect of LA on hemichannel activity and survival of HLE-B3 cells (a human lens epithelial cell line). We found that HLE-B3 cells expresses Cx43, Cx46, and Cx50 and can form functional hemichannels in their plasma membrane. The extracellular exposure to 10–50 μM of LA increases hemichannels activity (dye uptake) in a concentration-dependent manner, which was reduced by Cx-channel blockers, such as the Cx-mimetic peptide Gap27 and TATGap19, La3+, carbenoxolone (CBX) and the Akt kinase inhibitor. Additionally, LA increases intracellular calcium, which is attenuated in the presence of TATGap19, a specific Cx43-hemichannel inhibitor. Finally, the long exposure of HLE-B3 cells to LA 20 and 50 μM, reduced cell viability, which was prevented by CBX. Moreover, LA increased the proportion of apoptotic HLE-B3 cells, effect that was prevented by the Cx-mimetic peptide TAT-Gap19 but not by Akt inhibitor. Altogether, these findings strongly suggest a contribution of hemichannels opening in the cell death induced by LA in HLE-B3 cells. These cells can be an excellent tool to develop pharmacological studies in vitro.https://www.frontiersin.org/article/10.3389/fphys.2019.01574/fulllensconnexinpolyunsaturated fatty acidscell deathhemichannels |
| spellingShingle | Vania A. Figueroa Vania A. Figueroa Oscar Jara Carolina A. Oliva Marcelo Ezquer Fernando Ezquer Mauricio A. Retamal Mauricio A. Retamal Mauricio A. Retamal Agustín D. Martínez Guillermo A. Altenberg Guillermo A. Altenberg Aníbal A. Vargas Aníbal A. Vargas Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3) Frontiers in Physiology lens connexin polyunsaturated fatty acids cell death hemichannels |
| title | Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3) |
| title_full | Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3) |
| title_fullStr | Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3) |
| title_full_unstemmed | Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3) |
| title_short | Contribution of Connexin Hemichannels to the Decreases in Cell Viability Induced by Linoleic Acid in the Human Lens Epithelial Cells (HLE-B3) |
| title_sort | contribution of connexin hemichannels to the decreases in cell viability induced by linoleic acid in the human lens epithelial cells hle b3 |
| topic | lens connexin polyunsaturated fatty acids cell death hemichannels |
| url | https://www.frontiersin.org/article/10.3389/fphys.2019.01574/full |
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