TY - JOUR
T1 - Live imaging of targeted cell ablation in Xenopus
T2 - a new model to study demyelination and repair
AU - Kaya, Ferdinand
AU - Mannioui, Abdelkrim
AU - Chesneau, Albert
AU - Sekizar, Sowmya
AU - Maillard, Emmanuelle
AU - Ballagny, Chantal
AU - Houel-Renault, Ludivine
AU - Dupasquier, David
AU - Bronchain, Odile
AU - Holtzmann, Isabelle
AU - Desmazieres, Anne
AU - Thomas, Jean-Léon
AU - Demeneix, Barbara A
AU - Brophy, Peter J
AU - Zalc, Bernard
AU - Mazabraud, Andre
PY - 2012/9/12
Y1 - 2012/9/12
N2 - Live imaging studies of the processes of demyelination and remyelination have so far been technically limited in mammals. We have thus generated a Xenopus laevis transgenic line allowing live imaging and conditional ablation of myelinating oligodendrocytes throughout the CNS. In these transgenic pMBP-eGFP-NTR tadpoles the myelin basic protein (MBP) regulatory sequences, specific to mature oligodendrocytes, are used to drive expression of an eGFP (enhanced green fluorescent protein) reporter fused to the Escherichia coli nitroreductase (NTR) selection enzyme. This enzyme converts the innocuous prodrug metronidazole (MTZ) to a cytotoxin. Using two-photon imaging in vivo, we show that pMBP-eGFP-NTR tadpoles display a graded oligodendrocyte ablation in response to MTZ, which depends on the exposure time to MTZ. MTZ-induced cell death was restricted to oligodendrocytes, without detectable axonal damage. After cessation of MTZ treatment, remyelination proceeded spontaneously, but was strongly accelerated by retinoic acid. Altogether, these features establish the Xenopus pMBP-eGFP-NTR line as a novel in vivo model for the study of demyelination/remyelination processes and for large-scale screens of therapeutic agents promoting myelin repair.
AB - Live imaging studies of the processes of demyelination and remyelination have so far been technically limited in mammals. We have thus generated a Xenopus laevis transgenic line allowing live imaging and conditional ablation of myelinating oligodendrocytes throughout the CNS. In these transgenic pMBP-eGFP-NTR tadpoles the myelin basic protein (MBP) regulatory sequences, specific to mature oligodendrocytes, are used to drive expression of an eGFP (enhanced green fluorescent protein) reporter fused to the Escherichia coli nitroreductase (NTR) selection enzyme. This enzyme converts the innocuous prodrug metronidazole (MTZ) to a cytotoxin. Using two-photon imaging in vivo, we show that pMBP-eGFP-NTR tadpoles display a graded oligodendrocyte ablation in response to MTZ, which depends on the exposure time to MTZ. MTZ-induced cell death was restricted to oligodendrocytes, without detectable axonal damage. After cessation of MTZ treatment, remyelination proceeded spontaneously, but was strongly accelerated by retinoic acid. Altogether, these features establish the Xenopus pMBP-eGFP-NTR line as a novel in vivo model for the study of demyelination/remyelination processes and for large-scale screens of therapeutic agents promoting myelin repair.
KW - Xenopus laevis
KW - Animals
KW - Demyelinating Diseases
KW - Nerve Regeneration
KW - Humans
KW - Disease Models, Animal
KW - Microscopy, Fluorescence, Multiphoton
U2 - 10.1523/JNEUROSCI.2252-12.2012
DO - 10.1523/JNEUROSCI.2252-12.2012
M3 - Article
C2 - 22973012
SN - 1529-2401
VL - 32
SP - 12885
EP - 12895
JO - Journal of Neuroscience
JF - Journal of Neuroscience
IS - 37
ER -