Automated 3D immunofluorescence analysis of Dorsal Root Ganglia for the investigation of neural circuit alterations: a preliminary study.

Diabetic polyneuropathy is a major complication of diabetes mellitus, causing severe alterations of the neural circuits between spinal nerves and spinal cord. The analysis of 3D confocal images of dorsal root ganglia in diabetic mice, where different fluorescent markers are used to identify different types of nociceptors, can help understanding the unknown mechanisms of this pathology. Nevertheless, due to the inherent challenges of 3D confocal imaging, a thorough and comprehensive visual investigation is very difficult. In this work we introduce a tool, 3DRG, that provides a fully-automated segmentation and 3D rendering of positively labeled nociceptors in a dorsal root ganglion, as well a quantitative characterisation of its immunopositivity to each fluorescent marker. Our preliminary experiments on 3D confocal images of entire dorsal root ganglia from healthy and diabetic mice provided very interesting insights about the effects of the pathology on two different types of nociceptors.