Preganglionic sympathetic neurones innervating the rat adrenal medulla: immunocytochemical evidence of synaptic input from nerve terminals containing substance P, GABA or 5-hydroxytryptamine

Abstract

Sympathetic preganglionic neurones that innervate the adrenal medulla were identified for subsequent light and electron microscopic study by the retrograde transport of horseradish peroxidase (HRP) or a conjugate of HRP and cholera B-chain. Most labelled neurones were found in the intermediolateral column, but some occurred in the intercalated nucleus and in the lateral funiculus of the thoracic spinal cord. Three morphologically distinct types of neurone were retrogradely labelled, two of which had dendrites that extended medially towards the central canal and laterally across the entire lateral funiculus. A combination of retrograde labelling with pre-embedding immunocytochemistry allowed us to demonstrate synaptic contacts between boutons immunoreactive for substance P or 5-hydroxytryptamine (5-HT) and the cell bodies or proximal dendrites of sympathoadrenal neurones. The 5-HT-immunoreactive boutons appeared to be of two morphologically distinct types. Postembedding immunocytochemistry enabled us to show that sympathoadrenal neurones receive a heavy synaptic innervation from GABA-immunoreactive boutons: 32% of a random series of boutons in synaptic contact with cell bodies were GABA-immunoreactive. Proximal dendrites and also distal dendrites within the white matter were ensheathed in synaptic boutons, 37% of which were GABA-immunoreactive. It is concluded that sympathoadrenal neurones receive at least 4 distinct types of afferent synaptic input: from neurones containing substance P, or GABA and from two types of neurones containing 5-HT. The presence of synaptic inputs on distal dendrites that extend across the white matter adds further complexities to the control of the activity of sympathetic preganglionic neurones.