USING A 3D APPROACH TO DESCRIBE CELL POPULATIONS IN THE RAT DORSAL ROOT GANGLIA

Dorsal root ganglia (DRG) sensitive neurons represent the connection between the peripheral sensorial receptors and the central nervous system. These neurons are enwrapped individually by the satellite glial cells (SCGs) from which they receive metabolic support. Together, neuron and SCGs, become a functional unit that, in absence of the blood brain barrier, is easily exposed to external stress and damage insults. This intimate connection/relationship, both morphological and functional, can be partially pictured and studied following traditional slicing 2D histopathological techniques. Indeed, morphological cellular and subcellular alterations and changes in protein expression and/or distribution can be observed using classical techniques. However, a whole-3D approach avoids the serial sectioning required for quantitative results plus is able to show the cyto-architecture of the organ and a more complete picture of the anatomical relationship between cell populations close to physiological conditions. Here we use a 3D imaging technique to show the cyto-architecture of the DRG after “colouring” by immunofluorescence the different DRG cell populations and to assess alterations in DRG of neuropathic rats. CGRP, IB4 and MAP2 markers were useful to study the different neuronal populations. The IB4-MAP2 combination was able to label all neurons while the CGRP-IB4 couple could not but still both settings showed a small subpopulation of neurons where the proteins were co-expressed. Moreover, GFAP, ATF3 and connexin 43 were used as markers of damage in the DRG from neuropathic animals.