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The sequence of postsynaptic changes by iontophoresis. An early experiment of th type is shown in Fig. 2 Dennis, Harris & Kuffler, 1971 ; . This method aimed a * obtaining an ' artificial synapse' in which the nerve terminal could be replaced by a transmitter-filled pipette. With such an arrangement one could control quantity and timing of transmitter reaching the subsynaptic membrane without having to rely on the vagaries of the nerve impulse in the terminals. Eventually, with Doju Yoshikami we further improved the methodology and did more accurate analyses of synaptic events in skeletal neuromuscular junctions of the snake in which the motor axons at the endplate form terminals which in shape and fine structure closely resemble boutons'on neurones. In Fig. 3 we can compare neuronal and neuromuscular boutons. In Fig. 3 A the optics are focused on a rather prominent neuronal bouton, while 3 B shows a cross-section through such a bouton, with all the features of a chemical synapse. Figure 3 C gives a face-on view of an endplate in which one sees the bouton-like terminals embedded in the muscle surface in a ball-andsocket-like arrangement. A section through one of these boutons Fig. 3D ; again displays the characteristics of a chemical neuronal synapse, such as mitochondria and synaptic vesicles facing a synaptic cleft. The inset in Fig. 3 D ; shows a micrograph of a pipette whose tip opening was about 500 A. The tips of such pipettes can be placed accurately at the edge of a synaptic cleft for the release of ACh onto the subsynaptic membrane. Such pipette-release can rather faithfully reproduce in size and time course left, Fig. 4 ; a miniature synaptic potential caused by a spontaneous nerve-released quantum of transmitter right, Fig. 4 ; . The number of ACh molecules that mimicked the action of a quantum was determined with an accuracy of several thousand. This showed jhat on the average fewer than 10000 molecules of ACh produced quantal effects. Since the number of quanta causing an e.p.s.p. could be measured, the method provided quantitative details about the amount of transmitter released by nerve impulses Kuffler & Yoshikami, 1975 a, b ; . The endplate of the snake also gave clues about the kinetics of transmitter-receptor activation Hartzell, KufBer & Yoshikami, 1975 ; . The experiments on the cardiac ganglia of the frog and the nerve-muscle junctions in the snake were quite satisfactory, in particular for bringing into line some essential elements in neuromuscular and neuronal synaptic transmission. They affirmed once more the general validity of the earlier studies on fast e.p.s.p.s in skeletal muscles, particularly those by Katz, Fatt, del Castillo, Miledi and their colleagues. Yet the approach fell short of some of our objectives, which included inhibitory transmission and slow processes at synapses. The study of such events in vertebrates previously had been confined mainly to sympathetic ganglia, as we shall see. Much of our basic information, however, derives from molluscan neurones, particularly in Aplysia, which have continued to reveal many novel features of neuronal mechanisms that are then frequently found in vertebrates Tauc, 1967; Parnas & Strumwasser, 1974; Ascher & Kehoe, 1975; Gerschenfeld, 1977 ; . Around the time when we were searching for a more suitable preparation to explore various slow synaptic signals, McMahan & Purves 1976 ; made a fortunate discovery in the parasympathetic cardiac ganglion of the mudpuppy Necturus maculosus ; , another amphibian. Unexpectedly, the organization of this parasympathetic ganglion differed from that in the interatrial septum of the frog, because not only did it contafl the anticipated relatively large regular cholinergic ganglion cells principal cells and buspar.

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