Measuring endogenous levels of unconjugated bilirubin released from isolated murine brain tissue during oxygen-glucose deprivation

Han-Wei Liu; Ke Lai; Li-Na Gong; Hai-Bo Shi; Shan-Kai Yin; Lu-Yang Wang

STAR Protocols (Sep 2023)

Measuring endogenous levels of unconjugated bilirubin released from isolated murine brain tissue during oxygen-glucose deprivation

Han-Wei Liu,
Ke Lai,
Li-Na Gong,
Hai-Bo Shi,
Shan-Kai Yin,
Lu-Yang Wang

Affiliations

Han-Wei Liu: Department of Otorhinolaryngology Head & Neck Surgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China; Corresponding author
Ke Lai: Department of Otorhinolaryngology Head & Neck Surgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China; Program in Neuroscience and Mental Health, SickKids Research Institute, Toronto, ON M5G 1X8, Canada; Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada
Li-Na Gong: Department of Otorhinolaryngology Head & Neck Surgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
Hai-Bo Shi: Department of Otorhinolaryngology Head & Neck Surgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
Shan-Kai Yin: Department of Otorhinolaryngology Head & Neck Surgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
Lu-Yang Wang: Program in Neuroscience and Mental Health, SickKids Research Institute, Toronto, ON M5G 1X8, Canada; Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Corresponding author

Journal volume & issue: Vol. 4, no. 3
p. 102550

Abstract

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Summary: Quantitative assessment of endogenously synthesized and released bilirubin from brain tissue remains a challenge. Here, we present a sensitive and reproducible experimental paradigm to quantify, in real time, unconjugated bilirubin (UCB) from isolated murine brain tissue during oxygen-glucose deprivation (OGD). We describe steps for perfusion, brain dissection, brain slice preparation and incubation, glucose depletion, and OGD processing. We then detail procedures for standard calibration plotting and sample UCB measurement.For complete details on the use and execution of this protocol, please refer to Liu et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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