Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development

Authors

Nghi M. Nguyen, University of Nebraska Medical Center
Daniel Meyer, University of Nebraska Medical Center
Luke Meyer, University of Nebraska Medical Center
Subhash Chand, University of Nebraska Medical CenterFollow
Jagadesan Sankarasubramanian, University of Nebraska Medical CenterFollow
Maireen Miravite, University of Nebraska Medical CenterFollow
Chittibabu Guda, University of Nebraska Medical CenterFollow
Sowmya V. Yelamanchili, University of Nebraska Medical CenterFollow
Gurudutt Pendyala, University of Nebraska Medical CenterFollow

Document Type

Article

Journal Title

Cells

Publication Date

2023

Volume

12

Abstract

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.

MeSH Headings

Midazolam, Proteome, Hypnotics and Sedatives, Brain, Models, Biological

DOI Link

10.3390/cells12060966

ISSN

2073-4409

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Nguyen, Nghi M.; Meyer, Daniel; Meyer, Luke; Chand, Subhash; Sankarasubramanian, Jagadesan; Miravite, Maireen; Guda, Chittibabu; Yelamanchili, Sowmya V.; and Pendyala, Gurudutt, "Identification of YWHAH as a Novel Brain-Derived Extracellular Vesicle Marker Post Long-Term Midazolam Exposure during Early Development" (2023). Journal Articles: Anesthesiology. 16.
https://digitalcommons.unmc.edu/com_anesth_articles/16