Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development
Recently, the long-term use of sedative agents in the neonatal intensive care unit (NICU) has raised concerns about neurodevelopmental outcomes in exposed neonates. Midazolam (MDZ), a common neonatal sedative in the NICU, has been suggested to increase learning disturbances and cognitive impairment...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/6/966 |
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| author | Nghi M. Nguyen Daniel Meyer Luke Meyer Subhash Chand Sankarasubramanian Jagadesan Maireen Miravite Chittibabu Guda Sowmya V. Yelamanchili Gurudutt Pendyala |
| author_facet | Nghi M. Nguyen Daniel Meyer Luke Meyer Subhash Chand Sankarasubramanian Jagadesan Maireen Miravite Chittibabu Guda Sowmya V. Yelamanchili Gurudutt Pendyala |
| author_sort | Nghi M. Nguyen |
| collection | DOAJ |
| description | Recently, the long-term use of sedative agents in the neonatal intensive care unit (NICU) has raised concerns about neurodevelopmental outcomes in exposed neonates. Midazolam (MDZ), a common neonatal sedative in the NICU, has been suggested to increase learning disturbances and cognitive impairment in children. However, molecular mechanisms contributing to such outcomes with long-term MDZ use during the early stages of life remain unclear. In this study, we for the first time elucidate the role of brain-derived extracellular vesicles (BDEVs), including mining the BDEV proteome post long-term MDZ exposure during early development. Employing our previously established rodent model system that mimics the exposure of MDZ in the NICU using an increasing dosage regimen, we isolated BDEVs from postnatal 21-days-old control and MDZ groups using a differential sucrose density gradient. BDEVs from the control and MDZ groups were then characterized using a ZetaView nanoparticle tracking analyzer and transmission electron microscopy analysis. Next, using RT-qPCR, we examined the expression of key ESCRT-related genes involved in EV biogenesis. Lastly, using quantitative mass spectrometry-based proteomics, we mined the BDEV protein cargo that revealed key differentially expressed proteins and associated molecular pathways to be altered post long-term MDZ exposure. Our study characterized the proteome in BDEV cargo from long-term MDZ exposure at early development. Importantly, we identified and validated the expression of YWHAH as a potential target for further characterization of its downstream mechanism and a potential biomarker for the early onset of neurodevelopment and neurodegenerative diseases. Overall, the present study demonstrated long-term exposure to MDZ at early development stages could influence BDEV protein cargo, which potentially impact neural functions and behavior at later stages of development. |
| first_indexed | 2024-03-11T06:47:54Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T06:47:54Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-af8dc9bc55304faaa55319481fe9cbcc2023-11-17T10:14:25ZengMDPI AGCells2073-44092023-03-0112696610.3390/cells12060966Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early DevelopmentNghi M. Nguyen0Daniel Meyer1Luke Meyer2Subhash Chand3Sankarasubramanian Jagadesan4Maireen Miravite5Chittibabu Guda6Sowmya V. Yelamanchili7Gurudutt Pendyala8Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Genetics, Cell Biology, and Anatomy, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Genetics, Cell Biology, and Anatomy, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USADepartment of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USARecently, the long-term use of sedative agents in the neonatal intensive care unit (NICU) has raised concerns about neurodevelopmental outcomes in exposed neonates. Midazolam (MDZ), a common neonatal sedative in the NICU, has been suggested to increase learning disturbances and cognitive impairment in children. However, molecular mechanisms contributing to such outcomes with long-term MDZ use during the early stages of life remain unclear. In this study, we for the first time elucidate the role of brain-derived extracellular vesicles (BDEVs), including mining the BDEV proteome post long-term MDZ exposure during early development. Employing our previously established rodent model system that mimics the exposure of MDZ in the NICU using an increasing dosage regimen, we isolated BDEVs from postnatal 21-days-old control and MDZ groups using a differential sucrose density gradient. BDEVs from the control and MDZ groups were then characterized using a ZetaView nanoparticle tracking analyzer and transmission electron microscopy analysis. Next, using RT-qPCR, we examined the expression of key ESCRT-related genes involved in EV biogenesis. Lastly, using quantitative mass spectrometry-based proteomics, we mined the BDEV protein cargo that revealed key differentially expressed proteins and associated molecular pathways to be altered post long-term MDZ exposure. Our study characterized the proteome in BDEV cargo from long-term MDZ exposure at early development. Importantly, we identified and validated the expression of YWHAH as a potential target for further characterization of its downstream mechanism and a potential biomarker for the early onset of neurodevelopment and neurodegenerative diseases. Overall, the present study demonstrated long-term exposure to MDZ at early development stages could influence BDEV protein cargo, which potentially impact neural functions and behavior at later stages of development.https://www.mdpi.com/2073-4409/12/6/96614-3-3 etaMidazolamNICUextracellular vesiclesproteomicsneurodevelopment |
| spellingShingle | Nghi M. Nguyen Daniel Meyer Luke Meyer Subhash Chand Sankarasubramanian Jagadesan Maireen Miravite Chittibabu Guda Sowmya V. Yelamanchili Gurudutt Pendyala Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development Cells 14-3-3 eta Midazolam NICU extracellular vesicles proteomics neurodevelopment |
| title | Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development |
| title_full | Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development |
| title_fullStr | Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development |
| title_full_unstemmed | Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development |
| title_short | Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development |
| title_sort | identification of ywhah as a novel brain derived extracellular vesicle marker post long term midazolam exposure during early development |
| topic | 14-3-3 eta Midazolam NICU extracellular vesicles proteomics neurodevelopment |
| url | https://www.mdpi.com/2073-4409/12/6/966 |
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