HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina
A major player in the homeostatic response to hypoxia is the hypoxia-inducible factor (HIF)-1 that transactivates a number of genes involved in neovessel proliferation in response to low oxygen tension. In the retina, hypoxia overstimulates β-adrenoceptors (β-ARs) which play a key role in the format...
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MDPI AG
2022-04-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/11/8/1271 |
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| author | Rosario Amato Francesco Pisani Emiliano Laudadio Maurizio Cammalleri Martina Lucchesi Silvia Marracci Luca Filippi Roberta Galeazzi Maria Svelto Massimo Dal Monte Paola Bagnoli |
| author_facet | Rosario Amato Francesco Pisani Emiliano Laudadio Maurizio Cammalleri Martina Lucchesi Silvia Marracci Luca Filippi Roberta Galeazzi Maria Svelto Massimo Dal Monte Paola Bagnoli |
| author_sort | Rosario Amato |
| collection | DOAJ |
| description | A major player in the homeostatic response to hypoxia is the hypoxia-inducible factor (HIF)-1 that transactivates a number of genes involved in neovessel proliferation in response to low oxygen tension. In the retina, hypoxia overstimulates β-adrenoceptors (β-ARs) which play a key role in the formation of pathogenic blood vessels. Among β-ARs, β3-AR expression is increased in proliferating vessels in concomitance with increased levels of HIF-1α and vascular endothelial growth factor (VEGF). Whether, similarly to VEGF, hypoxia-induced β3-AR upregulation is driven by HIF-1 is still unknown. We used the mouse model of oxygen-induced retinopathy (OIR), an acknowledged model of retinal angiogenesis, to verify the hypothesis of β3-AR transcriptional regulation by HIF-1. Investigation of β3-AR regulation over OIR progression revealed that the expression profile of β3-AR depends on oxygen tension, similar to VEGF. The additional evidence that HIF-1α stabilization decouples β3-AR expression from oxygen levels further indicates that HIF-1 regulates the expression of the β3-AR gene in the retina. Bioinformatics predicted the presence of six HIF-1 binding sites (HBS #1-6) upstream and inside the mouse β3-AR gene. Among these, HBS #1 has been identified as the most suitable HBS for HIF-1 binding. Chromatin immunoprecipitation-qPCR demonstrated an effective binding of HIF-1 to HBS #1 indicating the existence of a physical interaction between HIF-1 and the β3-AR gene. The additional finding that β3-AR gene expression is concomitantly activated indicates the possibility that HIF-1 transactivates the β3-AR gene. Our results are indicative of β3-AR involvement in HIF-1-mediated response to hypoxia. |
| first_indexed | 2024-03-09T11:02:13Z |
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| id | doaj.art-720760ebddf14d408563dd8e1cca9240 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T11:02:13Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-720760ebddf14d408563dd8e1cca92402023-12-01T01:13:02ZengMDPI AGCells2073-44092022-04-01118127110.3390/cells11081271HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse RetinaRosario Amato0Francesco Pisani1Emiliano Laudadio2Maurizio Cammalleri3Martina Lucchesi4Silvia Marracci5Luca Filippi6Roberta Galeazzi7Maria Svelto8Massimo Dal Monte9Paola Bagnoli10Department of Biology, University of Pisa, 56127 Pisa, ItalyDepartment of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari Aldo Moro, 70125 Bari, ItalyDepartment of Materials, Environmental Sciences and Urban Planning, Polytechnic University of Marche, 60131 Ancona, ItalyDepartment of Biology, University of Pisa, 56127 Pisa, ItalyDepartment of Biology, University of Pisa, 56127 Pisa, ItalyDepartment of Biology, University of Pisa, 56127 Pisa, ItalyDepartment of Clinical and Experimental Medicine, Division of Neonatology and NICU, University of Pisa, 56100 Pisa, ItalyDepartment of Life and Environmental Sciences, Polytechnic University of Marche, 60131 Ancona, ItalyDepartment of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari Aldo Moro, 70125 Bari, ItalyDepartment of Biology, University of Pisa, 56127 Pisa, ItalyDepartment of Biology, University of Pisa, 56127 Pisa, ItalyA major player in the homeostatic response to hypoxia is the hypoxia-inducible factor (HIF)-1 that transactivates a number of genes involved in neovessel proliferation in response to low oxygen tension. In the retina, hypoxia overstimulates β-adrenoceptors (β-ARs) which play a key role in the formation of pathogenic blood vessels. Among β-ARs, β3-AR expression is increased in proliferating vessels in concomitance with increased levels of HIF-1α and vascular endothelial growth factor (VEGF). Whether, similarly to VEGF, hypoxia-induced β3-AR upregulation is driven by HIF-1 is still unknown. We used the mouse model of oxygen-induced retinopathy (OIR), an acknowledged model of retinal angiogenesis, to verify the hypothesis of β3-AR transcriptional regulation by HIF-1. Investigation of β3-AR regulation over OIR progression revealed that the expression profile of β3-AR depends on oxygen tension, similar to VEGF. The additional evidence that HIF-1α stabilization decouples β3-AR expression from oxygen levels further indicates that HIF-1 regulates the expression of the β3-AR gene in the retina. Bioinformatics predicted the presence of six HIF-1 binding sites (HBS #1-6) upstream and inside the mouse β3-AR gene. Among these, HBS #1 has been identified as the most suitable HBS for HIF-1 binding. Chromatin immunoprecipitation-qPCR demonstrated an effective binding of HIF-1 to HBS #1 indicating the existence of a physical interaction between HIF-1 and the β3-AR gene. The additional finding that β3-AR gene expression is concomitantly activated indicates the possibility that HIF-1 transactivates the β3-AR gene. Our results are indicative of β3-AR involvement in HIF-1-mediated response to hypoxia.https://www.mdpi.com/2073-4409/11/8/1271oxygen-induced retinopathyHIF-1 binding sitecomputational analysisChIP-qPCRgene expression |
| spellingShingle | Rosario Amato Francesco Pisani Emiliano Laudadio Maurizio Cammalleri Martina Lucchesi Silvia Marracci Luca Filippi Roberta Galeazzi Maria Svelto Massimo Dal Monte Paola Bagnoli HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina Cells oxygen-induced retinopathy HIF-1 binding site computational analysis ChIP-qPCR gene expression |
| title | HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina |
| title_full | HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina |
| title_fullStr | HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina |
| title_full_unstemmed | HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina |
| title_short | HIF-1-Dependent Induction of β3 Adrenoceptor: Evidence from the Mouse Retina |
| title_sort | hif 1 dependent induction of β3 adrenoceptor evidence from the mouse retina |
| topic | oxygen-induced retinopathy HIF-1 binding site computational analysis ChIP-qPCR gene expression |
| url | https://www.mdpi.com/2073-4409/11/8/1271 |
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