Analysis of cellular mechanisms of acute injuries in the central and peripheral nervous system using novel imaging approaches

Abstract

For analysis of the progression of regeneration in the central and peripheral nervous system usually are used techniques from histological sections that provide a two-dimensional image of the regenerating axons, showing only fragments, and may thus lead to misinterpretations. In order to overcome this obstacle, the tetrahydrofuran (THF)-based clearing technique was proposed. Through this technique, the brain stem and spinal cords of rodents become transparent allowing the deep-tissue imaging of axons. Since this technique is standardized only for spinal cord, this paper proposes the establishment of the technique for peripheral nerves. For this, we will use transgenic mice that express fluorescent proteins in neurons, astrocytes and microglia as well as double and triple hybrids. Microscopic analysis, by 2P-LSM will include the observation of cleared tissues (spinal cord and sciatic nerves) from transgenic mice before and after injury. (AU)