In Vivo Imaging of Anterograde and Retrograde Axonal Transport in Rodent Peripheral Nerves

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2020 Jun 12

PMID 32524487

Citations 17

Authors

James N Sleigh

Andrew P Tosolini

Giampietro Schiavo

Affiliations

Soon will be listed here.

Abstract

Axonal transport, which is the process mediating the active shuttling of a variety cargoes from one end of an axon to the other, is essential for the development, function, and survival of neurons. Impairments in this dynamic process are linked to diverse nervous system diseases and advanced ageing. It is thus essential that we quantitatively study the kinetics of axonal transport to gain an improved understanding of neuropathology as well as the molecular and cellular mechanisms regulating cargo trafficking. One of the best ways to achieve this goal is by imaging individual, fluorescent cargoes in live systems and analyzing the kinetic properties of their progression along the axon. We have therefore developed an intravital technique to visualize different organelles, such as signaling endosomes and mitochondria, being actively transported in the axons of both motor and sensory neurons in live, anesthetized rodents. In this chapter, we provide step-by-step instructions on how to deliver specific organelle-targeting, fluorescent probes using several routes of administration to image individual cargoes being bidirectionally transported along axons within the exposed sciatic nerve. This method can provide detailed, physiologically relevant information on axonal transport, and is thus poised to elucidate mechanisms regulating this process in both health and disease.

Citing Articles

The node of Ranvier influences the axonal transport of mitochondria and signaling endosomes.

Tosolini A, Abatecola F, Negro S, Sleigh J, Schiavo G iScience. 2024; 27(11):111158.

PMID: 39524336 PMC: 11544082. DOI: 10.1016/j.isci.2024.111158.

The tyrosine phosphatases LAR and PTPRδ act as receptors of the nidogen-tetanus toxin complex.

Surana S, Villarroel-Campos D, Rhymes E, Kalyukina M, Panzi C, Novoselov S EMBO J. 2024; 43(16):3358-3387.

PMID: 38977849 PMC: 11329502. DOI: 10.1038/s44318-024-00164-8.

Age-specific and compartment-dependent changes in mitochondrial homeostasis and cytoplasmic viscosity in mouse peripheral neurons.

Sleigh J, Mattedi F, Richter S, Annuario E, Ng K, Steinmark I Aging Cell. 2024; 23(10):e14250.

PMID: 38881280 PMC: 11464114. DOI: 10.1111/acel.14250.

Real-time imaging of axonal membrane protein life cycles.

Tyagi S, Higerd-Rusli G, Akin E, Baker C, Liu S, Dib-Hajj F Nat Protoc. 2024; 19(9):2771-2802.

PMID: 38831222 PMC: 11721981. DOI: 10.1038/s41596-024-00997-x.

Boosting BDNF in muscle rescues impaired axonal transport in a mouse model of DI-CMTC peripheral neuropathy.

Rhymes E, Simkin R, Qu J, Villarroel-Campos D, Surana S, Tong Y bioRxiv. 2024; .

PMID: 38559020 PMC: 10979848. DOI: 10.1101/2023.04.09.536152.

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