Nov 13, 2018

Development of an Acute Method to Deliver Transgenes Into the Brains of Adult Xenopus laevis

Frontiers in Neural Circuits
Ayako YamaguchiPaulo Rodrigues

Abstract

The central vocal pathway of the African clawed frog, Xenopus laevis, is a powerful vertebrate model to understand mechanisms underlying central pattern generation. However, fast and efficient methods of introducing exogenous genes into the neurons of adult X. laevis are currently not available. Here, we systematically tested methods of transgene delivery into adult X. laevis neurons. Although successfully used for tadpole neurons for over a decade, electroporation was not efficient in transfecting adult neurons. Similarly, adeno-associated virus (AAV) was not reliable, and lentivirus (LV) failed to function as viral vector in adult Xenopus neurons. In contrast, vesicular stomatitis virus (VSV) was a fast and robust vector for adult X. laevis neurons. Although toxic to the host cells, VSV appears to be less virulent to frog neurons than they are to mice neurons. At a single cell level, infected neurons showed normal physiological properties up to 7 days post infection and vocal circuits that included infected neurons generated normal fictive vocalizations up to 9 days post infection. The relatively long time window during which the physiology of VSV-infected neurons can be studied presents an ideal condition for the use of opto...Continue Reading

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Mentioned in this Paper

Vertebrates
Study
Amphibians
Biochemical Pathway
Subfamily lentivirinae
Presynaptic Terminals
Shuttle Vectors
Patterns
Research
Transfection

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