Electron microscopic and histochemical observations on different types of nerve endings in the extraocular muscles of the rat

Summary Nerve endings in the extraocular muscles of the rat were submitted to histochemical tests for formalin-induced fluorescence and carboxylic esterases. Acetylthiocholine, butyrylthiocholine and -naphthyl acetate were used as substrates and iso-OMPA, 284C51, eserine and E-600 as inhibitors. The ultrastructure of the endings was studied with the electron microscope.Both single and multiple nerve terminals were observed in all six extraocular muscles. The single terminals of myelinated axons were comparable in their light and electron microscopic structure with the typical motor end plates of other striated muscles, and like these they exhibit acetylcholinesterase (AChE), non-specific cholinesterase (ns. ChE) and non-specific esterase (ns. E) activity. These endings were apposed to twitch-type muscle fibres.The multiple terminals were classified with the light microscope into two types. The larger type was 1/3 of the size of the motor end plate; 2–5 endings innervated the same muscle fibre; subneural infoldings were weakly developed and possessed only slight AChE and ns. ChE and probably no ns. E activity. No subneural lamellae were visible under the light microscope in the smaller type, which also possessed AChE and ns. ChE and was composed of 10–20 small dots dispersed along a single muscle fibre. The Schwann cells along nerve fibres leading to these two types of multiple endings exhibited ns. ChE but not AChE and ns. E activity.The ultrastructure of the two types of multiple endings was principally similar. The main difference, compared with the motor end plate, was that these endings were derived from unmyelinated axons which either make synaptic contacts along their course with the muscle fibre at variable distances (smaller-type) or these terminals were grouped closely together (larger-type).A few dense-core vesicles were observed in these unmyelinated nerves and in their terminals which were considerably smaller than those in the motor end plate. They were not always separated from each other by sarcoplasm and teloglia (larger-type) and contained also empty vesicles. The secondary synaptic clefts were often sparse and irregular or even absent, but the typical myoneural postsynaptic electron density was always observed. These multiple endings, in contrast to the motor end plate, were apposed only to muscle fibres with slow contraction.No catecholamine containing nerve endings were observed in the extraocular muscles. These observations indicate that the rat extraocular muscles have a double cholinergic innervation.The author wishes to express his gratitude to Professor Antti Telkkä, M. D., Head of the Electron Microscope Laboratory, University of Helsinki, for permission to avail himself of the electron microscope facilities.