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

Nghi M. Nguyen; Daniel Meyer; Luke Meyer; Subhash Chand; Sankarasubramanian Jagadesan; Maireen Miravite; Chittibabu Guda; Sowmya V. Yelamanchili; Gurudutt Pendyala

doi:10.3390/cells12060966

Cells (Mar 2023)

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

Nghi M. Nguyen,
Daniel Meyer,
Luke Meyer,
Subhash Chand,
Sankarasubramanian Jagadesan,
Maireen Miravite,
Chittibabu Guda,
Sowmya V. Yelamanchili,
Gurudutt Pendyala

Affiliations

Nghi M. Nguyen: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Daniel Meyer: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Luke Meyer: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Subhash Chand: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Sankarasubramanian Jagadesan: Department of Genetics, Cell Biology, and Anatomy, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Maireen Miravite: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Chittibabu Guda: Department of Genetics, Cell Biology, and Anatomy, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Sowmya V. Yelamanchili: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA
Gurudutt Pendyala: Department of Anesthesiology, University of Nebraska Medical Center (UNMC), Omaha, NE 68198, USA

DOI: https://doi.org/10.3390/cells12060966
Journal volume & issue: Vol. 12, no. 6
p. 966

Abstract

Read online

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.

Published in Cells

ISSN: 2073-4409 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Biology (General): Cytology
Website: https://www.mdpi.com/journal/cells

About the journal

Abstract

Keywords