blood substitutes BUT only a few fibres regenerate Provide Growth Factors? NGF only works on peripheral nerves BDNF? Genetically engineered cells?
Improve the substrate? laminin osmotic pumps artificial substrates Change the cell? Engineer more receptors Up-regulate internal messengers use different cells?
Fetal cells Stem cells Will mammalian spinal cords make the right connections even if we do get regeneration? No retinotopy in lizard regeneration? No retinotopy in regeneration along sciatic nerve?
Retinal transplants do not form a retinotopic projection? What can and cant regenerate?
Adult fish can regenerate everything! Larval amphibia too, adults slowly? Reptiles slowly and without specificity Mammalian PNS Mammalian olfactory tract Mammalian nerves can only sprout abortively
Are fish different? Why study preamniotes? Can differences give clues? Can we do novel experiments? How do their spinal
cords regenerate? Why study Lower Vertebrates? They regenerate quickly and are simple to maintain. Embryos are readily available They have many tracts in common with mammals They show many of the growth factors, ECM
components and even collapsins found in mammals Two approaches have been used 1. Assuming similar processes act in all species use them as a model system for testing the ability of
fibres to regenerate under experimental conditions 2. To deliberately look for clues in the differences between the regenerating and non-regenerating
species How different are they? They grow throughout life Level of myelin inhibitory factor low in adult animals? (or localised?) Glial response to injury different? Extracellular environment more condusive to regeneration?
Are their neurons different? Added throughout life Less need for growth factors? Less cell death after axotomy Do NOT all regenerate
Inhibiting factors ARE present in fish CNS? Regenerating fibres prefer the PNS? (Bentley and Zottoli 1993) Ventral root Fish optic nerve not permissive for adult nerves
(Sivron et al 1994) cut site Is the balance different ? L1 NGF BNTF
ECM MIF-1 cell death MAG What does a regenerated spinal cord look like?
Which fibres regenerate? Do they grow back to their normal tracts? Do they find the correct targets? Is this process an extension of growth?
Can Spinal Axons tell Right from Left? R L R ?
RL Cut L R
? ? L The problem of order
Spinal cord in culture Direct cell heaters Work well in closed systems Ideal for imaging as they allow DIC and Fluorescence concurrently Small amounts of media Allow long term culture
Only heat cells Must use objective heater with immersion optics Closed stage system Thermocouple
inlet drain Contacts for glass heaters Phase and Dark Field Artificial Substrates
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