Axonal mRNAs: characterisation and role in the growth and regeneration of dorsal root ganglion axons and growth cones.
We have developed a compartmentalised culture model for the purification of axonal mRNA from embryonic, neonatal and adult rat dorsal root ganglia. This mRNA was used un-amplified for RT-qPCR. We assayed for the presence of axonal mRNAs encoding molecules known to be involved in axon growth and guidance. mRNAs for beta-actin, beta-tubulin, and several molecules involved in the control of actin dynamics and signalling during axon growth were found, but mRNAs for microtubule-associated proteins, integrins and cell surface adhesion molecules were absent. Quantification of beta-actin mRNA by means of qPCR showed that the transcript is present at the same level in embryonic, newborn and adult axons. Using the photoconvertible reporter Kaede we showed that there is local translation of beta-actin in axons, the rate being increased by axotomy. Knock down of beta-actin mRNA by RNAi inhibited the regeneration of new axon growth cones after in vitro axotomy, indicating that local translation of actin-related molecules is important for successful axon regeneration.
Vogelaar
Christina F
CF
Cambridge University Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Robinson Way, Cambridge CB2 0PY, UK.
Gervasi
Noreen M
NM
Cambridge University Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Robinson Way, Cambridge CB2 0PY, UK.
Gumy
Laura F
LF
Cambridge University Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Robinson Way, Cambridge CB2 0PY, UK.
Story
David J
DJ
Cambridge University Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Robinson Way, Cambridge CB2 0PY, UK.
Raha-Chowdhury
Ruma
R
Cambridge University Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Robinson Way, Cambridge CB2 0PY, UK.
Leung
Kin-Mei
KM
Department of Physiology Development and Neuroscience, University of Cambridge, Cambridge CB2 3DY, UK.
Holt
Christine E
CE
Department of Physiology Development and Neuroscience, University of Cambridge, Cambridge CB2 3DY, UK.
Fawcett
James W
JW
Cambridge University Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Robinson Way, Cambridge CB2 0PY, UK.
D000199:Actins / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D001369:Axons / Q000502:physiology*; D019771:Axotomy; D003599:Cytoskeleton / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology; D020439:Growth Cones / Q000502:physiology*; D009416:Nerve Regeneration / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction / Q000235:genetics; D046509:Tissue Culture Techniques
D000199:Actins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface