학술논문
Spinal cord regeneration in Xenopus laevis
Document Type
Report
Author
Source
Nature Protocols. February 2017, Vol. 12 Issue 2, p372, 18 p.
Subject
Language
English
ISSN
1754-2189
Abstract
Author(s): Gabriela Edwards-Faret [1]; Rosana Muñoz [1]; Emilio E Méndez-Olivos [1]; Dasfne Lee-Liu [1]; Victor S Tapia [1]; Juan Larraín (corresponding author) [1] Introduction Spinal cord injury leads to permanent [...]
Here we present a protocol for the husbandry of Xenopus laevis tadpoles and froglets, and procedures to study spinal cord regeneration. This includes methods to induce spinal cord injury (SCI); DNA and morpholino electroporation for genetic studies; in vivo imaging for cell analysis; a swimming test to measure functional recovery; and a convenient model for screening for new compounds that promote neural regeneration. These protocols establish X. laevis as a unique model organism for understanding spinal cord regeneration by comparing regenerative and nonregenerative stages. This protocol can be used to understand the molecular and cellular mechanisms involved in nervous system regeneration, including neural stem and progenitor cell (NSPC) proliferation and neurogenesis, extrinsic and intrinsic mechanisms involved in axon regeneration, glial response and scar formation, and trophic factors. For experienced personnel, husbandry takes 1-2 months; SCI can be achieved in 5-15 min; and swimming recovery takes 20-30 d. Keywords: Xenopus laevis, spinal cord regeneration, neural stem/progenitor cell, NSPCs, neurogenesis, Sox2, spinal cord, electroporation, in vivo imaging, tadpole, froglet, frog, morpholino
Here we present a protocol for the husbandry of Xenopus laevis tadpoles and froglets, and procedures to study spinal cord regeneration. This includes methods to induce spinal cord injury (SCI); DNA and morpholino electroporation for genetic studies; in vivo imaging for cell analysis; a swimming test to measure functional recovery; and a convenient model for screening for new compounds that promote neural regeneration. These protocols establish X. laevis as a unique model organism for understanding spinal cord regeneration by comparing regenerative and nonregenerative stages. This protocol can be used to understand the molecular and cellular mechanisms involved in nervous system regeneration, including neural stem and progenitor cell (NSPC) proliferation and neurogenesis, extrinsic and intrinsic mechanisms involved in axon regeneration, glial response and scar formation, and trophic factors. For experienced personnel, husbandry takes 1-2 months; SCI can be achieved in 5-15 min; and swimming recovery takes 20-30 d. Keywords: Xenopus laevis, spinal cord regeneration, neural stem/progenitor cell, NSPCs, neurogenesis, Sox2, spinal cord, electroporation, in vivo imaging, tadpole, froglet, frog, morpholino