Cellular NeuroscienceResearch PaperConditioning lesions enhance growth state only in sensory neurons lacking calcitonin gene-related peptide and isolectin B4-binding
Section snippets
Surgical procedures
All experiments were performed in accordance with the Code of Practice for the Care and Use of Animals for Experimental Purposes (National Health and Medical Research Council of Australia) and approved by the Animal Care and Ethics Committee of the University of Sydney and Royal North Shore Hospital. These guidelines comply with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. All efforts were made to minimize the number of animals used and their suffering. A
Effect of prior sciatic nerve CCI on neurite growth in different populations of DRG neurons in vitro
Unilateral sciatic nerve transection or CCI was performed 7 days prior to culturing the L3–L5 DRG ipsilateral and contralateral to injury, which supply the sciatic nerve in mice (Rigaud et al., 2008). Cultures were grown overnight in either neurotrophin-free medium or NGF-supplemented medium. Neurite initiation and elongation were measured in CGRP-positive and IB4-binding neurons, which represent peptidergic and nonpeptidergic nociceptors, respectively (Fig. 1A–D). Neurons that were neither
Discussion
In this study, we have compared the regenerative growth state after a partial and complete conditioning lesion, sciatic nerve CCI and transection, in three neurochemically defined populations of DRG neurons. We showed that both CCI and transection activated a mode of sparsely branched, elongating neurite growth but this was restricted to a subset of nonCGRP/nonIB4-binding neurons; it did not occur in CGRP-positive or IB4-binding neurons. This indicates that a lower regenerative capacity
Conclusion
The results of this study indicated that specific types of DRG neurons differ in their ability to activate “elongating” regenerative growth, independent of the type of conditioning lesion. In addition, NGF does not stimulate an elongating mode of growth after nerve injury, but promotes a more linearly restricted, “arborizing” mode of growth which may be insufficient for long distance regenerative growth. These results suggests that some types of DRG neurons have a low intrinsic regenerative
Acknowledgments
National Health and Medical Research Council of Australia (Senior Research Fellowship358709); New South Wales (NSW) Office for Science and Medical Research “Spinal Cord Injury and Other Neurological Conditions” Program Grant; University of Sydney Medical Foundation; Australian Postgraduate Award (to A.K.). We thank Ian Napier for advising on surgical procedures.
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2015, Experimental NeurologyCitation Excerpt :These results indicate that in this model system, large and small diameter neurons regenerate equally well and replating neurons does not cause a selective loss of either major neuron type. Previous reports have shown that small nociceptive neurons have impaired regeneration (Kalous and Keast, 2010; Leclere et al., 2007). In order to test whether different neuronal subtypes activate the pro-regenerative state after dissociation, we classified neurons as being NF200 positive (large diameter mechano- and proprioreceptors) or NF200 negative (enriched for small diameter nociceptors) and assessed neurite length (Fig. S1).
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Current address: Department of Physiology, Development and Neurscience, Anatomy Building, University of Cambridge, Downing Street, Cambridge, CB2 3DY, UK.