Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells
Extracellular ATP and trophic factors released by exocytosis modulate in vivo proliferation, migration, and differentiation in multipotent stem cells (MpSC); however, the purinoceptors mediating this signaling remain uncharacterized in stem cells derived from the human olfactory epithelium (hOE). Ou...
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| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/2728786 |
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| author | Héctor Solís-Chagoyán Edgar Flores-Soto Marcela Valdés-Tovar Montserrat G. Cercós Eduardo Calixto Luis M. Montaño Carlos Barajas-López Bettina Sommer Arnoldo Aquino-Gálvez Citlali Trueta Gloria A. Benítez-King |
| author_facet | Héctor Solís-Chagoyán Edgar Flores-Soto Marcela Valdés-Tovar Montserrat G. Cercós Eduardo Calixto Luis M. Montaño Carlos Barajas-López Bettina Sommer Arnoldo Aquino-Gálvez Citlali Trueta Gloria A. Benítez-King |
| author_sort | Héctor Solís-Chagoyán |
| collection | DOAJ |
| description | Extracellular ATP and trophic factors released by exocytosis modulate in vivo proliferation, migration, and differentiation in multipotent stem cells (MpSC); however, the purinoceptors mediating this signaling remain uncharacterized in stem cells derived from the human olfactory epithelium (hOE). Our aim was to determine the purinergic pathway in isolated human olfactory neuronal precursor cells (hONPC) that exhibit MpSC features. Cloning by limiting dilution from a hOE heterogeneous primary culture was performed to obtain a culture predominantly constituted by hONPC. Effectiveness of cloning to isolate MpSC-like precursors was corroborated through immunodetection of specific protein markers and by functional criteria such as self-renewal, proliferation capability, and excitability of differentiated progeny. P2 receptor expression in hONPC was determined by Western blot, and the role of these purinoceptors in the ATP-induced exocytosis and changes in cytosolic Ca2+ ([Ca2+]i) were evaluated using the fluorescent indicators FM1-43 and Fura-2 AM, respectively. The clonal culture was enriched with SOX2 and OCT3/4 transcription factors; additionally, the proportion of nestin-immunopositive cells, the proliferation capability, and functionality of differentiated progeny remained unaltered through the long-term clonal culture. hONPC expressed P2X receptor subtypes 1, 3-5, and 7, as well as P2Y2, 4, 6, and 11; ATP induced both exocytosis and a transient [Ca2+]i increase predominantly by activation of metabotropic P2Y receptors. Results demonstrated for the first time that ex vivo-expressed functional P2 receptors in MpSC-like hONPC regulate exocytosis and Ca2+ signaling. This purinergic-triggered release of biochemical messengers to the extracellular milieu might be involved in the paracrine signaling among hOE cells. |
| format | Article |
| id | doaj-art-f26cef8732144c1495461bd154f9b1b8 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-f26cef8732144c1495461bd154f9b1b82025-02-03T01:28:47ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/27287862728786Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor CellsHéctor Solís-Chagoyán0Edgar Flores-Soto1Marcela Valdés-Tovar2Montserrat G. Cercós3Eduardo Calixto4Luis M. Montaño5Carlos Barajas-López6Bettina Sommer7Arnoldo Aquino-Gálvez8Citlali Trueta9Gloria A. Benítez-King10Instituto Nacional de Psiquiatría Ramón de la Fuente Muñiz, Laboratorio de Neurofarmacología, Calzada México-Xochimilco 101, San Lorenzo Huipulco, CP 14370 Ciudad de México, MexicoUniversidad Nacional Autónoma de México, Departamento de Farmacología, Facultad de Medicina, CP 04510 Ciudad de México, MexicoInstituto Nacional de Psiquiatría Ramón de la Fuente Muñiz, Laboratorio de Neurofarmacología, Calzada México-Xochimilco 101, San Lorenzo Huipulco, CP 14370 Ciudad de México, MexicoInstituto Nacional de Psiquiatría Ramón de la Fuente Muñiz, Departamento de Neurofisiología, Calzada México-Xochimilco 101, San Lorenzo Huipulco, CP 14370 Ciudad de México, MexicoInstituto Nacional de Psiquiatría Ramón de la Fuente Muñiz, Departamento de Neurobiología, Ciudad de México, MexicoUniversidad Nacional Autónoma de México, Departamento de Farmacología, Facultad de Medicina, CP 04510 Ciudad de México, MexicoInstituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, Col. Lomas 4ª Sección, CP 78216 San Luis Potosí, MexicoInstituto Nacional de Enfermedades Respiratorias, Departamento de Investigación en Hiperreactividad Bronquial, CP 14080 Ciudad de México, MexicoInstituto Nacional de Enfermedades Respiratorias, Laboratorio de Oncología Biomédica, CP 14080 Ciudad de México, MexicoInstituto Nacional de Psiquiatría Ramón de la Fuente Muñiz, Departamento de Neurofisiología, Calzada México-Xochimilco 101, San Lorenzo Huipulco, CP 14370 Ciudad de México, MexicoInstituto Nacional de Psiquiatría Ramón de la Fuente Muñiz, Laboratorio de Neurofarmacología, Calzada México-Xochimilco 101, San Lorenzo Huipulco, CP 14370 Ciudad de México, MexicoExtracellular ATP and trophic factors released by exocytosis modulate in vivo proliferation, migration, and differentiation in multipotent stem cells (MpSC); however, the purinoceptors mediating this signaling remain uncharacterized in stem cells derived from the human olfactory epithelium (hOE). Our aim was to determine the purinergic pathway in isolated human olfactory neuronal precursor cells (hONPC) that exhibit MpSC features. Cloning by limiting dilution from a hOE heterogeneous primary culture was performed to obtain a culture predominantly constituted by hONPC. Effectiveness of cloning to isolate MpSC-like precursors was corroborated through immunodetection of specific protein markers and by functional criteria such as self-renewal, proliferation capability, and excitability of differentiated progeny. P2 receptor expression in hONPC was determined by Western blot, and the role of these purinoceptors in the ATP-induced exocytosis and changes in cytosolic Ca2+ ([Ca2+]i) were evaluated using the fluorescent indicators FM1-43 and Fura-2 AM, respectively. The clonal culture was enriched with SOX2 and OCT3/4 transcription factors; additionally, the proportion of nestin-immunopositive cells, the proliferation capability, and functionality of differentiated progeny remained unaltered through the long-term clonal culture. hONPC expressed P2X receptor subtypes 1, 3-5, and 7, as well as P2Y2, 4, 6, and 11; ATP induced both exocytosis and a transient [Ca2+]i increase predominantly by activation of metabotropic P2Y receptors. Results demonstrated for the first time that ex vivo-expressed functional P2 receptors in MpSC-like hONPC regulate exocytosis and Ca2+ signaling. This purinergic-triggered release of biochemical messengers to the extracellular milieu might be involved in the paracrine signaling among hOE cells.http://dx.doi.org/10.1155/2019/2728786 |
| spellingShingle | Héctor Solís-Chagoyán Edgar Flores-Soto Marcela Valdés-Tovar Montserrat G. Cercós Eduardo Calixto Luis M. Montaño Carlos Barajas-López Bettina Sommer Arnoldo Aquino-Gálvez Citlali Trueta Gloria A. Benítez-King Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells Stem Cells International |
| title | Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells |
| title_full | Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells |
| title_fullStr | Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells |
| title_full_unstemmed | Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells |
| title_short | Purinergic Signaling Pathway in Human Olfactory Neuronal Precursor Cells |
| title_sort | purinergic signaling pathway in human olfactory neuronal precursor cells |
| url | http://dx.doi.org/10.1155/2019/2728786 |
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