Acetylcholinesterase and Acetylcholine Receptor Histochemistry on End Plate Regions, Myotendinous Junctions, and Sarcolemma in the Axial Musculature of Three Teleost Fish Species

In this paper we describe a rapid procedure for the identification of motor end plates in fish. We demonstrated the presence of acetylcholine receptors by means of an immune fluorescence technique with alpha-bungarotoxin. Koelle's thiocholine method was used to localize acetylcholinesterase (AChE) activity. Under carefully controlled conditions the AChE activity and the anti-alpha-bungarotoxin fluorescence showed an equal distribution. This means that in the study of motor innervation in fish the AChE reaction can be used to stain only the motor end plates, leaving the AChE rich preterminal axons unstained. Comparison of the AChE reaction pattern with the distribution of binding sites for antibodies raised against neurofilament protein revealed that in end plate regions high concentrations of AChE are only present in axons and end plates. The myotendinous junctions also possess a high receptor density and enzyme activity. A low enzymatic activity was found at the non-junctional periphery of white muscle fibres. This activity probably resides in the sarcolemma. No non-specific cholinesterase activity was found. From light microscopical analysis it appeared that a single end plate may innervate 2 adjacent muscle fibres. This was affirmed by ultrastructural observations. The dual innervation suggests that, in fish, motor units have a limited distribution through the myotome.