In Vivo Imaging of Axonal Organelle Transport in the Mouse Brain

Visualization and analysis of axonal organelle transport has been mostly conducted in vitro, using primary neuronal cell cultures, although more recently, intravital organelle imaging has been established in model organisms such as drosophila, zebrafish, and mouse. In this chapter, we describe a met...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2431; p. 95
Main Authors Knabbe, Johannes, Protzmann, Jil, Kuner, Thomas
Format Journal Article
LanguageEnglish
Published United States 2022
Subjects
Animals
Axonal Transport - physiology
Axons - metabolism
Brain
Drosophila
Mice
Organelles - metabolism
Zebrafish
Axonal transport
Cranial window surgery
In vivo imaging
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Summary:Visualization and analysis of axonal organelle transport has been mostly conducted in vitro, using primary neuronal cell cultures, although more recently, intravital organelle imaging has been established in model organisms such as drosophila, zebrafish, and mouse. In this chapter, we describe a method to visualize axonal transport of cellular organelles such as dense core vesicles or mitochondria in the living mouse brain in order to study organelle transport in its native environment. We achieve this goal by injecting adeno-associated viruses expressing fluorescently tagged marker proteins into thalamic nuclei of mice, thereby transducing neurons that project to the surface of the brain. Axonal projections and trafficking of organelles can be imaged with a 2-photon microscope through a chronically implanted window in the mouse skull in anesthetized as well as awake mice.
ISSN:1940-6029
DOI:10.1007/978-1-0716-1990-2_5