Soma neurofilament immunoreactivity is related to cell size and fibre conduction velocity in rat primary sensory neurons.

Abstract

1. Intracellular recordings were made in dorsal root ganglia in vitro at 37 degrees C. The L4, L5 and L6 ganglia from 46- to 51-day-old female Wistar rats were used. In each neuron conduction velocity (CV) was measured and fluorescent dye was injected. Later the intensity of the immunoreactivity to RT97 (a monoclonal antibody to the phosphorylated 200 kDa neurofilament subunit) as well as the cell size (cross-sectional area at the nuclear level) were measured in the dye-injected neurons. RT97 was used to distinguish between the L (light, neurofilament-rich) and the SD (small dark, neurofilament-poor) neuronal somata. 2. Neurons were classified as C neurons (CV less than 1.3 m/s), C/A delta neurons (1.3-2 m/s), A delta neurons (2-12 m/s) or A alpha/beta neurons (greater than 12 m/s). 3. All A-fibre somata were RT97 positive (L) and all C-fibre somata were RT97 negative (SD), although in the C/A delta group both positive and negative neurons were seen. Thus, RT97-negative somata had C (unmyelinated) or C/A delta fibres, while RT97-positive somata had A (myelinated) or C/A delta fibres. 4. The size distributions of A neurons and C neurons were consistent with their classification as L- and SD-cell neurons respectively. The size distribution of A delta cells was skewed with a peak of small cells and a tail of medium-sized cells. 5. There was a loose positive correlation between cell size and fibre CV. 6. RT97 intensity was positively correlated with CV if all neurons were considered together, but no correlation was seen within the C, A delta or A alpha/beta CV groups. 7. RT97 intensity was positively correlated with cell size when all neurons were considered together. Although no correlation was seen within the C or the A delta CV groups, a clear positive correlation was seen for A alpha/beta neurons. 8. The relationship of RT97 intensity to cell size was not demonstrably altered by axotomy, time in vitro or the presence of intracellular dye in control experiments. 9. RT97-negative and -positive neurons could be seen in neonatal rat ganglia. Their size distributions resembled, respectively, the SD- and L-neuron populations at this age. RT97 immunoreactivity may therefore be a useful predictor of the cell type and myelinated state which a sensory cell is destined to reach in the adult rat.