The main leg extensor muscle of the crayfish Procambarus clarkii is innervated by two excitatory motor axons (phasic and tonic) and by the common inhibitory axon. The axons and their terminals were investigated using vital fluorescent dyes, antibodies and electron microscopy. Correlative physiological observations were made using intracellular microelectrodes and focal 'macro-patch' electrodes. The excitatory axons innervate the muscle fibres more extensively than does the inhibitory axon. Striking morphological and physiological differences between the two excitatory axons were discerned. The tonic motor axon contains many mitochondria and has varicose junctional terminals; the phasic axon has fewer mitochondria and thin terminals. In freely moving animals, the tonic axon is very active, whereas the phasic axon is inactive but fires in brief bursts during walking. Throughout the muscle, tonic excitatory postsynaptic potentials (EPSPs) are very small at low frequencies and facilitate greatly at higher frequencies. The EPSPs of the phasic axon are larger at low frequencies and depress with maintained stimulation. At identified tonic terminals, quantal content at 1­10 Hz is very low, whereas at identified phasic terminals, quantal content is 50- to 200-fold greater. The results indicate that transmitter release is regulated differently at the synapses of the two axons.

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