Reinnervation of frog sympathetic ganglia with somatic nerve fibres

Abstract

The formation of synapses in the last two ganglia of the sympathetic chain of the frog Rana esculenta was investigated after anastomosing the 6th spinal nerve to the denervated ganglia in order to evaluate the reinnervation of deafferented sympathetic neurons with somatic cholinergic axons. The same ganglia were examined both electrophysiologically and morphologically from 25 to 280 days after the operation. In response to electrical stimulations of the anastomosed spinal nerve, synaptic transmission was analysed with intracellular microelectrodes placed into B or C sympathetic neurons. Synaptic density was quantified using electron microscopy by a synaptic index defined as the ratio of the number of synapses encountered to the number of perykarya examined. After ganglionic deafferentation, post-synaptic membrane differentiations persisted without any pre-synaptic element and an index of the ‘vacated’ post-synaptic differentiations was calculated. Although somatic axons were growing into all ganglia studied, no sign of neuronal reinnervation was detected in ganglia of 8 of the 31 frogs (26%) taken from 29 to 210 days after the anastomosis. Moreover, in 18 out of 31 frogs (58%) analysed at different times after the operation, the ganglia were reinnervated with regenerating preganglionic axons in spite of care taken to avoid it. However, even after 3 months, certain neurons of these ganglia were not reinnervated and the synaptic index approximated the value of normal ganglia only in the 8th ganglion. In addition, post-synaptic membrane differentiations could still persist and coexist with normal synapses. It was only beyond three months after the anastomosis that the ganglia of 5 of the 31 frogs (16%) were reinnervated with regenerating somatic axons. Reinnervated B and C neurons were polyinnervated. But in 3 out of these 5 frogs the ganglia were also reinnervated with preganglionic axons and several B and C neurons received a double reinnervation. The synaptic indices were far from the value of normal ganglia except for the 8th ganglion of one frog reinnervated by both types of axons and the indices of vacated differentiations were close to that of ganglia with no reinnervated neurons. Contrary to mammals, frog somatic axons are, therefore, relatively ineffective at reinnervating sympathetic neurons, probably because in amphibian ganglia, synapses between the pre- and post-synaptic elements require higher specificity.