Adult Mouse Retina Explants: From ex vivo to in vivo Model of Central Nervous System Injuries
Résumé
In mammals, adult neurons fail to regenerate following any insult to adult central nervous system (CNS), which leads to a permanent and irreversible loss of motor and cognitive functions. For a long time, much effort has been deployed to uncover mechanisms of axon regeneration in the CNS. Even if some cases of functional recovery have been reported, there is still a discrepancy regarding the functionality of a neuronal circuit upon lesion. Today, there is a need not only to identify new molecules implicated in adult CNS axon regeneration, but also to decipher the fine molecular mechanisms associated with regeneration failure. Here, we propose to use cultures of adult retina explants to study all molecular and cellular mechanisms that occur during CNS regeneration. We show that adult retinal explant cultures have the advantages to (i) recapitulate all the features observed in vivo , including axon regeneration induced by intrinsic factors, and (ii) be an ex vivo set-up with high accessibility and many downstream applications. Thanks to several examples, we demonstrate that adult explants can be used to address many questions, such as axon guidance, growth cone formation and cytoskeleton dynamics. Using laser guided ablation of a single axon, axonal injury can be performed at a single axon level, which allows to record early and late molecular events that occur after the lesion. Our model is the ideal tool to study all molecular and cellular events that occur during CNS regeneration at a single-axon level, which is currently not doable in vivo . It is extremely valuable to address unanswered questions of neuroprotection and neuroregeneration in the context of CNS lesion and neurodegenerative diseases.
Mots clés
central nervous system
axon regeneration
explants
ex vivo
optic nerve
retinal ganglion cells
growth cone
axonal transport AAV2
Adeno-associated virus type 2 BSA
bovine serum albumin CNS
central nervous system CNTF
ciliary neurotrophic factor Cre
Cre recombinase CTB-555
Alexa 555-conjugated cholera toxin B DCLK2
doublecortin like kinase 2 GFP
green fluorescent protein JAK/STAT
Janus Kinase/Signal Transducer and Activator of Transcription KLF
krüppel like factors mTOR
mammalian target of rapamycin PBS
phosphate buffered saline PFA
paraformaldehyde Plap
placental alkaline phosphatase PNS
peripheral nervous system PTEN
Phosphatase and TENsin homolog PTEN
Adeno-associated virus type 2
BSA
bovine serum albumin
CNS
central nervous system
CNTF
ciliary neurotrophic factor
Cre
Cre recombinase
CTB-555
Alexa 555-conjugated cholera toxin B
DCLK2
doublecortin like kinase 2
GFP
green fluorescent protein
JAK/STAT
Janus Kinase/Signal Transducer and Activator of Transcription
KLF
krüppel like factors
mTOR
mammalian target of rapamycin
PBS
phosphate buffered saline
PFA
paraformaldehyde
Plap
placental alkaline phosphatase
PNS
peripheral nervous system
PTEN
Phosphatase and TENsin homolog
Domaines
Sciences du Vivant [q-bio]| Origine | Fichiers éditeurs autorisés sur une archive ouverte |
|---|