Direct Injection of a Lentiviral Vector Highlights Multiple Motor Pathways in the Rat Spinal Cord

Introducing proteins of interest into cells in the nervous system is challenging due to innate biological barriers that limit access to most molecules. Injection directly into spinal cord tissue bypasses these barriers, providing access to cell bodies or synapses where molecules can be incorporated....

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Bibliographic Details
Published in:Journal of visualized experiments no. 145
Main Authors: Keefe, Kathleen M., Junker, Ian P., Sheikh, Imran S., Campion, Thomas J., Smith, George M.
Format: Journal Article
Language:English
Published: United States 15.03.2019
Subjects:
Animals
axons
brain stem
Brain Stem - metabolism
Efferent Pathways - physiology
Female
gene therapy
Genetic Vectors - administration & dosage
green fluorescent protein
Injections
interneurons
Lentivirus - genetics
Lumbar Vertebrae - metabolism
Neurons - metabolism
rats
Rats, Sprague-Dawley
spinal cord
Spinal Cord - metabolism
synapse
Transduction, Genetic
viruses
ISSN:1940-087X, 1940-087X
Online Access:Get full text
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Summary:Introducing proteins of interest into cells in the nervous system is challenging due to innate biological barriers that limit access to most molecules. Injection directly into spinal cord tissue bypasses these barriers, providing access to cell bodies or synapses where molecules can be incorporated. Combining viral vector technology with this method allows for introduction of target genes into nervous tissue for the purpose of gene therapy or tract tracing. Here a virus engineered for highly efficient retrograde transport (HiRet) is introduced at the synapses of propriospinal interneurons (PNs) to encourage specific transport to neurons in the spinal cord and brainstem nuclei. Targeting PNs takes advantage of the numerous connections they receive from motor pathways such as the rubrospinal and reticulospinal tracts, as well as their interconnection with each other throughout spinal cord segments. Representative tracing using the HiRet vector with constitutively active green fluorescent protein (GFP) shows high fidelity details of cell bodies, axons and dendritic arbors in thoracic PNs and in reticulospinal neurons in the pontine reticular formation. HiRet incorporates well into brainstem pathways and PNs but shows age dependent integration into corticospinal tract neurons. In summary, spinal cord injection using viral vectors is a suitable method for introduction of proteins of interest into neurons of targeted tracts.
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ISSN:1940-087X
1940-087X
DOI:10.3791/59160