aconitine and biocytin

The aim of the present study was to determine whether, in young rats, spinal motoneurons possess functional nicotinic acetylcholine receptors. Motoneurons were identified either by retrograde labelling or by choline acetyltransferase immunohistochemistry. Whole-cell recordings were performed in spinal cord slices cut at the lumbar level. In voltage clamp, acetylcholine evoked a rapidly activating inward current. In current clamp, it depolarized the motoneuron membrane and induced action potential firing. The acetylcholine-evoked current was strongly reduced by d-tubocurarine or dihydro-beta-erythroidine, broad spectrum nicotinic antagonists, but was almost insensitive to methyllycaconitine, a nicotinic antagonist selective for receptors containing the alpha7 subunit. Moreover, exo-2-(2-pyridyl)-7-azabicyclo[2.2.1]heptane, an alpha7-specific agonist, was without effect. In young animals, light-microscopic autoradiography showed that in the central grey matter all laminae were intensely and equally labelled by [3H]epibatidine. A dense [125I]-alpha-bungarotoxin binding was also found in all laminae, with slightly lower levels in the superficial layers of the dorsal horns and in the ventral part of the grey matter. In adults, the density of [3H]epibatidine binding sites was much lower in the entire grey matter, except in layer 2 of the dorsal horn, and [125I]-alpha-bungarotoxin binding sites were present only in some selected areas. Our data indicate that spinal motoneurons possess functional nicotinic receptors of the heteromeric type and suggest that nicotinic cholinergic transmission may play a significant role in the developing spinal cord.

Topics: Acetylcholine; Aconitine; Age Factors; Animals; Animals, Newborn; Autoradiography; Binding Sites; Bridged Bicyclo Compounds, Heterocyclic; Bungarotoxins; Choline O-Acetyltransferase; Dihydro-beta-Erythroidine; Immunohistochemistry; In Vitro Techniques; Iodine Isotopes; Lysine; Membrane Potentials; Motor Neurons; Nicotinic Agonists; Nicotinic Antagonists; Patch-Clamp Techniques; Pyridines; Rats; Rats, Sprague-Dawley; Receptors, Nicotinic; Spinal Cord; Tritium; Tubocurarine