Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms
Purinergic signaling has been implicated in many biological functions, including development. In this study, we investigate the functions of extracellular adenosine and adenosine receptors using a mouse embryonic stem cell (ESC) line and morula stages isolated from mouse embryos. Feeder-free mouse E...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2023.1328398/full |
| _version_ | 1797351541097627648 |
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| author | Talita Glaser Patrícia Martins Renata Beco Carolina Adriane Bento Angelica R. Cappellari Sophia La Banca Oliveira Christian Albert Merkel Vanessa Fernandes Arnaud-Sampaio Claudiana Lameu Ana Maria Battastini Henning Ulrich |
| author_facet | Talita Glaser Patrícia Martins Renata Beco Carolina Adriane Bento Angelica R. Cappellari Sophia La Banca Oliveira Christian Albert Merkel Vanessa Fernandes Arnaud-Sampaio Claudiana Lameu Ana Maria Battastini Henning Ulrich |
| author_sort | Talita Glaser |
| collection | DOAJ |
| description | Purinergic signaling has been implicated in many biological functions, including development. In this study, we investigate the functions of extracellular adenosine and adenosine receptors using a mouse embryonic stem cell (ESC) line and morula stages isolated from mouse embryos. Feeder-free mouse ESC was investigated in the absence and presence of the leukemia inhibitory factor (LIF), configuring undifferentiated cells and cells undergoing spontaneous differentiation. High alkaline phosphatase (ALPL) and low CD73 levels resulting in low adenosine (eADO) levels were characteristic for pluripotent cells in the presence of the LIF, while LIF deprivation resulted in augmented adenosine levels and reduced pluripotency marker expression, which indicated differentiation. Tracing ESC proliferation by BrdU labeling revealed that the inhibition of ALPL by levamisole resulted in a decrease in proliferation due to less eADO accumulation. Furthermore, caffeine and levamisole treatment, inhibiting adenosine receptor and eADO accumulation, respectively, reduced ESC migration, similar to that observed in the absence of the LIF. Pharmacological approaches of selective adenosine receptor subtype inhibition triggered specific adenosine receptor activities, thus triggering calcium or MAP kinase pathways leading to differentiation. In line with the in vitro data, mouse embryos at the morula stage were sensitive to treatments with A1 and A3 receptor antagonists, leading to the conclusion that A1 receptor and A3 receptor inhibition impairs proliferation and self-renewal and triggers inappropriate differentiation, respectively. The findings herein define the functions of eADO signaling in early development with implications for developmental disorders, in which adenosine receptors or ectonucleotidase dysfunctions are involved, and which could lead to malformations and miscarriages, due to exposure to caffeine. |
| first_indexed | 2024-03-08T13:01:57Z |
| format | Article |
| id | doaj.art-1ef63b8d3b44427e955125d111cdd131 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-03-08T13:01:57Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-1ef63b8d3b44427e955125d111cdd1312024-01-19T04:51:16ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122024-01-011410.3389/fphar.2023.13283981328398Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanismsTalita Glaser0Patrícia Martins1Renata Beco2Carolina Adriane Bento3Angelica R. Cappellari4Sophia La Banca Oliveira5Christian Albert Merkel6Vanessa Fernandes Arnaud-Sampaio7Claudiana Lameu8Ana Maria Battastini9Henning Ulrich10Department of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Biochemistry, Federal University of Rio Grande do Sul, Porto Alegre, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Health (São Paulo—State), Medical School of the University of São Paulo (HCFMUSP), SãoPaulo, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilDepartment of Biochemistry, Federal University of Rio Grande do Sul, Porto Alegre, BrazilDepartment of Biochemistry, Institute of Chemistry, University of São Paulo, SãoPaulo, BrazilPurinergic signaling has been implicated in many biological functions, including development. In this study, we investigate the functions of extracellular adenosine and adenosine receptors using a mouse embryonic stem cell (ESC) line and morula stages isolated from mouse embryos. Feeder-free mouse ESC was investigated in the absence and presence of the leukemia inhibitory factor (LIF), configuring undifferentiated cells and cells undergoing spontaneous differentiation. High alkaline phosphatase (ALPL) and low CD73 levels resulting in low adenosine (eADO) levels were characteristic for pluripotent cells in the presence of the LIF, while LIF deprivation resulted in augmented adenosine levels and reduced pluripotency marker expression, which indicated differentiation. Tracing ESC proliferation by BrdU labeling revealed that the inhibition of ALPL by levamisole resulted in a decrease in proliferation due to less eADO accumulation. Furthermore, caffeine and levamisole treatment, inhibiting adenosine receptor and eADO accumulation, respectively, reduced ESC migration, similar to that observed in the absence of the LIF. Pharmacological approaches of selective adenosine receptor subtype inhibition triggered specific adenosine receptor activities, thus triggering calcium or MAP kinase pathways leading to differentiation. In line with the in vitro data, mouse embryos at the morula stage were sensitive to treatments with A1 and A3 receptor antagonists, leading to the conclusion that A1 receptor and A3 receptor inhibition impairs proliferation and self-renewal and triggers inappropriate differentiation, respectively. The findings herein define the functions of eADO signaling in early development with implications for developmental disorders, in which adenosine receptors or ectonucleotidase dysfunctions are involved, and which could lead to malformations and miscarriages, due to exposure to caffeine.https://www.frontiersin.org/articles/10.3389/fphar.2023.1328398/fullpurinergic signalingERK1/2calcium signalingembryonic stem cellsadenosinecaffeine |
| spellingShingle | Talita Glaser Patrícia Martins Renata Beco Carolina Adriane Bento Angelica R. Cappellari Sophia La Banca Oliveira Christian Albert Merkel Vanessa Fernandes Arnaud-Sampaio Claudiana Lameu Ana Maria Battastini Henning Ulrich Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms Frontiers in Pharmacology purinergic signaling ERK1/2 calcium signaling embryonic stem cells adenosine caffeine |
| title | Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms |
| title_full | Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms |
| title_fullStr | Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms |
| title_full_unstemmed | Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms |
| title_short | Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms |
| title_sort | impairment of adenosine signaling disrupts early embryo development unveiling the underlying mechanisms |
| topic | purinergic signaling ERK1/2 calcium signaling embryonic stem cells adenosine caffeine |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2023.1328398/full |
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