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Intrinsic properties inhibit axonal outgrowth from neonatal rat spinal cord explant
J-M P Mouveroux1, E A J F Lakke, E Marani
1Neuroregulation Group, Dept. of Neurosurgery, Leiden University Medical Center (LUMC), PO Box 9604, NL-2300 RC Leiden, The Netherlands. Mouveroux@lumc.nl
Insights
Neurite outgrowth from neonatal rat spinal cord explants decreases significantly with age. This age-related decline in nerve regeneration appears to be an intrinsic property of the explant.
Area of Science:
- Neuroscience
- Developmental Biology
- Regenerative Medicine
Background:
- Neonatal rat spinal cord explants are used to study neural development and regeneration.
- Previous studies have observed age-related decreases in neuronal outgrowth in various models.
Purpose of the Study:
- To investigate the effect of age at explantation on neurite outgrowth from lumbar spinal cord explants.
- To quantify the relationship between postnatal age and the number of outgrowing neurites.
Main Methods:
- Lumbar spinal cord explants from rat pups (postnatal day 0-6) were cultured on poly-ethylene-imine (PEI) and poly-D-lysin (PDL) coated surfaces.
- Neurite outgrowth was quantified and statistically analyzed in relation to the age of the explant.
- Control explants included lumbar explants in a collagen matrix and cortical explants.
Main Results:
- A significant inverse correlation was found between the age of the rat pup at explantation and the number of outgrowing neurites.
- The number of neurites decreased sharply with increasing postnatal age, following specific logarithmic expressions on PEI and PDL.
- Similar age-related decreases in outgrowth were observed in collagen matrix cultures.
Conclusions:
- The age of the neonatal rat spinal cord explant is a critical factor influencing neurite outgrowth.
- The observed phenomenon suggests an intrinsic growth inhibitory property of the explant that increases with age.
- This age-related decline in regenerative capacity may be related to changes at the interface between the explant and its environment.
Abstract:
Lumbar spinal cord explants, harvested from neonatal rat pups aged between postnatal day 0 (P0) and P6, were cultured for a period of 48 hrs in the chemically defined medium R(12) on a poly-ethylene-imine (PEI) and on poly-D-lysin (PDL) coated surface. The outgrowth outside the explant was quantified. Lumbar explants from the same rat and embedded in a collagen matrix, and cortical explants from a P0 rat were used as controls. Statistical analysis demonstrated a clear relation between age-at-explantation and the number of neurites in the corona surrounding the explant. The number of outgrowing neurites decreased sharply with age-at-explantation. The average number of neurites per explant obeyed to the expression log (n) = -0.736x + 3.294 on PEI, and log (n) = -0.721x + 2.295 on PDL; x epsilon in [P0 - P6] (n, the number of neurites per explant; x, the age-at-explantation expressed in postnatal days). A similar observed age-related decrease of outgrowth has been described when culturing the lumbar explant inside a collagen matrix. The phenomenon appears to be an intrinsic property of the explant. We review growth inhibitory properties in different models and propose that the phenomenon occurs here at the interface explant-world.
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