Assessment of BMP responses in an in vitro model of acute ethanol toxicity
The assay presented here was designed to assess the immediate effects of ethanol (EtOH) exposure on intracellular signaling activated by BMPs (Bone Morphogenetic Proteins). Previous reports of the relationship between EtOH exposure and BMP-dependent signaling have primarily assessed the expression o...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-01-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016122000164 |
| _version_ | 1811178776668667904 |
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| author | Naila Habeeb Sheyda Najafi Jeanette C. Perron |
| author_facet | Naila Habeeb Sheyda Najafi Jeanette C. Perron |
| author_sort | Naila Habeeb |
| collection | DOAJ |
| description | The assay presented here was designed to assess the immediate effects of ethanol (EtOH) exposure on intracellular signaling activated by BMPs (Bone Morphogenetic Proteins). Previous reports of the relationship between EtOH exposure and BMP-dependent signaling have primarily assessed the expression of individual BMPs, changes in BMP target genes or effects on the phosphorylation level of key downstream mediators after days or weeks of in vivo EtOH exposure. What happens to BMP-stimulated signaling immediately following exposure to EtOH remains largely unexplored. Here, the early events of BMP-evoked intracellular signaling were examined in an in vitro model of acute EtOH toxicity. The BMP/Ethanol Stimulation Assay involved first stimulating cultured cells with recombinant BMPs. BMP-evoked intracellular signaling was then allowed to develop for 30 minutes. Next, the cells were exposed to a range of EtOH concentrations for an additional 30 minutes. Finally, the cultures were processed for Western blot analysis or immunofluorescent labeling.This short-term assay:• Permits investigation of EtOH exposure during the initial signaling events downstream of BMP receptor activation• Enables assessment of how the presence of BMPs might protect against cellular injury caused by toxic EtOH levels |
| first_indexed | 2024-04-11T06:24:47Z |
| format | Article |
| id | doaj.art-21f74d63a38046ab91c0c81d71318a24 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-04-11T06:24:47Z |
| publishDate | 2022-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-21f74d63a38046ab91c0c81d71318a242022-12-22T04:40:25ZengElsevierMethodsX2215-01612022-01-019101631Assessment of BMP responses in an in vitro model of acute ethanol toxicityNaila Habeeb0Sheyda Najafi1Jeanette C. Perron2Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's UniversityDepartment of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's UniversityCorresponding author.; Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's UniversityThe assay presented here was designed to assess the immediate effects of ethanol (EtOH) exposure on intracellular signaling activated by BMPs (Bone Morphogenetic Proteins). Previous reports of the relationship between EtOH exposure and BMP-dependent signaling have primarily assessed the expression of individual BMPs, changes in BMP target genes or effects on the phosphorylation level of key downstream mediators after days or weeks of in vivo EtOH exposure. What happens to BMP-stimulated signaling immediately following exposure to EtOH remains largely unexplored. Here, the early events of BMP-evoked intracellular signaling were examined in an in vitro model of acute EtOH toxicity. The BMP/Ethanol Stimulation Assay involved first stimulating cultured cells with recombinant BMPs. BMP-evoked intracellular signaling was then allowed to develop for 30 minutes. Next, the cells were exposed to a range of EtOH concentrations for an additional 30 minutes. Finally, the cultures were processed for Western blot analysis or immunofluorescent labeling.This short-term assay:• Permits investigation of EtOH exposure during the initial signaling events downstream of BMP receptor activation• Enables assessment of how the presence of BMPs might protect against cellular injury caused by toxic EtOH levelshttp://www.sciencedirect.com/science/article/pii/S2215016122000164Western blotImmunofluorescent labelingSmadPI3K/AktC2C12 cellsDRG neurons |
| spellingShingle | Naila Habeeb Sheyda Najafi Jeanette C. Perron Assessment of BMP responses in an in vitro model of acute ethanol toxicity MethodsX Western blot Immunofluorescent labeling Smad PI3K/Akt C2C12 cells DRG neurons |
| title | Assessment of BMP responses in an in vitro model of acute ethanol toxicity |
| title_full | Assessment of BMP responses in an in vitro model of acute ethanol toxicity |
| title_fullStr | Assessment of BMP responses in an in vitro model of acute ethanol toxicity |
| title_full_unstemmed | Assessment of BMP responses in an in vitro model of acute ethanol toxicity |
| title_short | Assessment of BMP responses in an in vitro model of acute ethanol toxicity |
| title_sort | assessment of bmp responses in an in vitro model of acute ethanol toxicity |
| topic | Western blot Immunofluorescent labeling Smad PI3K/Akt C2C12 cells DRG neurons |
| url | http://www.sciencedirect.com/science/article/pii/S2215016122000164 |
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