The Role of the Primary Cilium in Sensing Extracellular pH
Biosensors on the membrane of the vascular endothelium are responsible for sensing mechanical and chemical signals in the blood. Transduction of these stimuli into intracellular signaling cascades regulate cellular processes including ion transport, gene expression, cell proliferation, and/or cell d...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-07-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/8/7/704 |
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| author | Kimberly F. Atkinson Rinzhin T. Sherpa Surya M. Nauli |
| author_facet | Kimberly F. Atkinson Rinzhin T. Sherpa Surya M. Nauli |
| author_sort | Kimberly F. Atkinson |
| collection | DOAJ |
| description | Biosensors on the membrane of the vascular endothelium are responsible for sensing mechanical and chemical signals in the blood. Transduction of these stimuli into intracellular signaling cascades regulate cellular processes including ion transport, gene expression, cell proliferation, and/or cell death. The primary cilium is a well-known biosensor of shear stress but its role in sensing extracellular pH change has never been examined. As a cellular extension into the immediate microenvironment, the cilium could be a prospective sensor for changes in pH and regulator of acid response in cells. We aim to test our hypothesis that the primary cilium plays the role of an acid sensor in cells using vascular endothelial and embryonic fibroblast cells as in vitro models. We measure changes in cellular pH using pH-sensitive 2′,7′-biscarboxyethy1-5,6-carboxyfluorescein acetoxy-methylester (BCECF) fluorescence and mitogen-activated protein kinase (MAPK) activity to quantify responses to both extracellular pH (pH<sub>o</sub>) and intracellular pH (pH<sub>i</sub>) changes. Our studies show that changes in pH<sub>o</sub> affect pH<sub>i</sub> in both wild-type and cilia-less <i>Tg737</i> cells and that the kinetics of the pH<sub>i</sub> response are similar in both cells. Acidic pH<sub>o</sub> or pH<sub>i</sub> was observed to change the length of primary cilia in wild-type cells while the cilia in <i>Tg737</i> remained absent. Vascular endothelial cells respond to acidic pH through activation of ERK1/2 and p38-mediated signaling pathways. The cilia-less <i>Tg737</i> cells exhibit delayed responsiveness to pH<sub>o</sub> dependent and independent pH<sub>i</sub> acidification as depicted in the phosphorylation profile of ERK1/2 and p38. Otherwise, intracellular pH homeostatic response to acidic pH<sub>o</sub> is similar between wild-type and <i>Tg737</i> cells, indicating that the primary cilia may not be the sole sensor for physiological pH changes. These endothelial cells respond to pH changes with a predominantly K<sup>+</sup>-dependent pH<sub>i</sub> recovery mechanism, regardless of ciliary presence or absence. |
| first_indexed | 2024-03-12T06:22:30Z |
| format | Article |
| id | doaj.art-ef60a5065c7d47c1a3e5bdd563daadfc |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T06:22:30Z |
| publishDate | 2019-07-01 |
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| series | Cells |
| spelling | doaj.art-ef60a5065c7d47c1a3e5bdd563daadfc2023-09-03T02:09:20ZengMDPI AGCells2073-44092019-07-018770410.3390/cells8070704cells8070704The Role of the Primary Cilium in Sensing Extracellular pHKimberly F. Atkinson0Rinzhin T. Sherpa1Surya M. Nauli2Department of Biomedical & Pharmaceutical Sciences, Chapman University, Irvine, CA 92618, USADepartment of Biomedical & Pharmaceutical Sciences, Chapman University, Irvine, CA 92618, USADepartment of Biomedical & Pharmaceutical Sciences, Chapman University, Irvine, CA 92618, USABiosensors on the membrane of the vascular endothelium are responsible for sensing mechanical and chemical signals in the blood. Transduction of these stimuli into intracellular signaling cascades regulate cellular processes including ion transport, gene expression, cell proliferation, and/or cell death. The primary cilium is a well-known biosensor of shear stress but its role in sensing extracellular pH change has never been examined. As a cellular extension into the immediate microenvironment, the cilium could be a prospective sensor for changes in pH and regulator of acid response in cells. We aim to test our hypothesis that the primary cilium plays the role of an acid sensor in cells using vascular endothelial and embryonic fibroblast cells as in vitro models. We measure changes in cellular pH using pH-sensitive 2′,7′-biscarboxyethy1-5,6-carboxyfluorescein acetoxy-methylester (BCECF) fluorescence and mitogen-activated protein kinase (MAPK) activity to quantify responses to both extracellular pH (pH<sub>o</sub>) and intracellular pH (pH<sub>i</sub>) changes. Our studies show that changes in pH<sub>o</sub> affect pH<sub>i</sub> in both wild-type and cilia-less <i>Tg737</i> cells and that the kinetics of the pH<sub>i</sub> response are similar in both cells. Acidic pH<sub>o</sub> or pH<sub>i</sub> was observed to change the length of primary cilia in wild-type cells while the cilia in <i>Tg737</i> remained absent. Vascular endothelial cells respond to acidic pH through activation of ERK1/2 and p38-mediated signaling pathways. The cilia-less <i>Tg737</i> cells exhibit delayed responsiveness to pH<sub>o</sub> dependent and independent pH<sub>i</sub> acidification as depicted in the phosphorylation profile of ERK1/2 and p38. Otherwise, intracellular pH homeostatic response to acidic pH<sub>o</sub> is similar between wild-type and <i>Tg737</i> cells, indicating that the primary cilia may not be the sole sensor for physiological pH changes. These endothelial cells respond to pH changes with a predominantly K<sup>+</sup>-dependent pH<sub>i</sub> recovery mechanism, regardless of ciliary presence or absence.https://www.mdpi.com/2073-4409/8/7/704acidosisERK1/2p38pHprimary cilia |
| spellingShingle | Kimberly F. Atkinson Rinzhin T. Sherpa Surya M. Nauli The Role of the Primary Cilium in Sensing Extracellular pH Cells acidosis ERK1/2 p38 pH primary cilia |
| title | The Role of the Primary Cilium in Sensing Extracellular pH |
| title_full | The Role of the Primary Cilium in Sensing Extracellular pH |
| title_fullStr | The Role of the Primary Cilium in Sensing Extracellular pH |
| title_full_unstemmed | The Role of the Primary Cilium in Sensing Extracellular pH |
| title_short | The Role of the Primary Cilium in Sensing Extracellular pH |
| title_sort | role of the primary cilium in sensing extracellular ph |
| topic | acidosis ERK1/2 p38 pH primary cilia |
| url | https://www.mdpi.com/2073-4409/8/7/704 |
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