Neuromodulators Enhance Transmitter Release by Two Separate Mechanisms at the Inhibitor of Crayfish Opener Muscle

A presynaptic voltage control method has been used to investigate the modulatory effects of serotonin (5-HT) and okadaic acid (OA) on the inhibitory junction of the crayfish opener muscle. Instead of using action potentials, we used 20 msec pulses depolarized to 0 mV to activate transmitter release. This approach allowed us to monitor two separate physiological parameters related to the release process. The first parameter, transmitter release kinetics, is characterized as the delay when inhibitory postsynaptic conductance reaches its half-maximum (IPSG50). The second parameter, the total area of IPSG (IPSGarea), estimates total transmitter output. We have reported previously that the F2 component of synaptic facilitation is associated with a decrease in IPSG50 but without a change in IPSGarea. These results raised the possibility that IPSG50 and IPSGarea could be mediated by separate mechanisms that were modulated independently. To explore this possibility, we investigated the effects of 5-HT (100–200 nm) and OA (2.5 μm) on the two parameters. 5-HT and OA enhanced IPSG neither by changing the sensitivity of postsynaptic receptors, as tested by iontophoretically ejected GABA, nor by elevating resting and action potential-activated presynaptic free calcium, as monitored by fura-2 imaging. 5-HT and OA decreased IPSG50 by 3.0 ± 1.4 and 3.6 ± 1.1 msec, respectively, and increased IPSGarea by 50 ± 21 and 37 ± 6%, respectively. The ability of F2 facilitation to accelerate release kinetics was reduced in the presence of the modulators, suggesting that the mechanism underlying the accelerated release kinetics was shared by the two modes of synaptic enhancement. This report demonstrates that the acceleration in release kinetics and the increase in total release are two separate mechanisms for enhancing transmitter output and that these two mechanisms can be activated without changes in presynaptic calcium dynamics.