Work

Intravital Imaging of Fluorescent Protein Expression in Mice with a Closed-Skull Traumatic Brain Injury and Cranial Window Using a Two-Photon Microscope

Year: 2023

Type: article

Abstract: The goal of this protocol is to demonstrate how to longitudinally visualize the expression and localization of a protein of interest within specific cell types of an animal's brain, upon exposure to exogenous stimuli. Here, the administration of a closed-skull traumatic brain injury (TBI) and simultaneous implantation of a cranial window for subsequent longitudinal intravital imaging in mice is shown. Mice are intracranially injected with an adeno-associated virus (AAV) expressing enhanced green fluorescent protein (EGFP) under a neuronal specific promoter. After 2 to 4 weeks, the mice are subjected to a repetitive TBI using a weight drop device over the AAV injection location. Within the same surgical session, the mice are implanted with a metal headpost and then a glass cranial window over the TBI impacting site. The expression and cellular localization of EGFP is examined using a two-photon microscope in the same brain region exposed to trauma over the course of months.

Source: Journal of Visualized Experiments

DOI: 10.3791/64701

Authors: Jianjun Zhong, Georgia Gunner, Nils Henninger, Dorothy P. Schafer, Daryl A. Bosco

Institutions: University of Massachusetts Chan Medical School, The Affiliated Yongchuan Hospital of Chongqing Medical University

Language: English

Topic: Traumatic Brain Injury and Neurovascular Disturbances

Subfield: Neurology

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: green

Funders: U.S. Department of Defense, National Institute of Allergy and Infectious Diseases, National Institute of Neurological Disorders and Stroke