Assessment of unmyelinated fibre regeneration by the detection of axonal labelling of membrane-associated glycoconjugates in peripheral nerves by laser scanning confocal microscopy.

Overview

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One of the unique features of the peripheral nervous system (PNS) is the ability of its neurones to regenerate following a traumatic injury.  For example, following a complete transection of a peripheral nerve the damaged sensory neurones will regrow towards their peripheral target tissues and in some instances make functional contacts within these tissues.   Unfortunately this ability is greatly attenuated in neurones of the central nervous system (CNS) which means the distressing consequences of the brain and spinal cord lesions are, for the most part, irreversible. Clearly a detailed understanding of the mechanisms which enable PNS neurones to regenerate may provide insight into mechanisms by which CNS neurones may be encouraged to regrow.  This will hopefully lead to the development of more effective treatment strategies for brain and spinal cord damage. The aim of this project is to take advantage of some recent technological developments to establish a technique which allows the monitoring of the rate of axonal regrowth in the PNS following injury. The technique should prove to be a powerful tool in the study of axonal regeneration.

Approaches

Adult rats will be deeply anaesthetised with sodium pentobarbitone and (using aseptic procedures) a small incision made to expose either the saphenous nerve or the sural nerve, bilaterally. The exposed nerves will then be crushed in a carefully-controlled manner using a pair of haemostats cooled in liquid nitrogen. Between one week and three months following the nerve lesions the animals will be euthanased, the lesioned nerves removed and then screened for the presence of a unique galactose-containing glycoprotein expressed selectively by unmyelinated primary sensory neurones.  The presence of the glycoprotein will be detected using a fluorescent carbohydrate-binding protein and laser scanning confocal microscopy. Using this technique it will be possible to map the time course of the regeneration of unmyelinated primary sensory neurones and provide an accurate and quantifiable means for investigating the mechanisms which underlie regeneration in the peripheral nervous system.

References

  1. Thornton, P.J., Gerke, M.B. and Plenderleith, M.B. (2005) Histochemical localisation of a galactose-containing glycoconjugate expressed by sensory neurones innervating different peripheral tissues. Journal of the Peripheral Nervous System. 10: 47-57.
  2. Plenderleith, M.B. and Snow, P.J (1993) The plant lectin Bandeiraea simplicifolia I-B4 identifies a subpopulation of small diameter primary sensory neurones which innervate the skin in the rat. Neuroscience Letters. 159: 17-20.
Study level
Honours
Supervisors
QUT
Organisational unit

Science and Engineering Faculty

Research area

Medical Sciences

Contact
Please contact the supervisor.