Regulation of exocytosis by protein kinase C.

PKC (protein kinase C) has been known for many years to modulate regulated exocytosis in a wide variety of cell types. In neurons and neuroendocrine cells, PKC regulates several different stages of the exocytotic process, suggesting that these multiple actions of PKC are mediated by phosphorylation of distinct protein targets. In recent years, a variety of exocytotic proteins have been identified as PKC substrates, the best characterized of which are SNAP-25 (25 kDa synaptosome-associated protein) and Munc18. In the present study, we review recent evidence suggesting that site-specific phosphorylation of SNAP-25 and Munc18 by PKC regulates distinct stages of exocytosis.

Role for calcium in heat shock-mediated synaptic thermoprotection in Drosophila larvae.

Chemical synaptic transmission is the mechanism for fast, excitation-coupled information transfer between neurons. Previous work in larval Drosophila has shown that transmission at synaptic boutons is protected by heat shock exposure from subsequent thermal stress through pre- and postsynaptic modifications. This protective effect has been, at least partially, ascribed to an up-regulation in the inducible heat shock protein, hsp70. Effects of hsp70 are correlated with changes to intracellular calcium handling, and the dynamics of intracellular calcium regulate synaptic transmission. Consistent with such a relationship, synaptic plasticity increases at locust neuromuscular junctions following heat shock, suggesting an effect of heat shock on residual presynaptic calcium. Intracellular recording from single abdominal muscle fibers of Drosophila larvae showed that prior heat shock imparts thermoprotection by increasing the upper temperature limit for synaptic transmission. Heat shock exposure enhances short-term synaptic plasticity and increases its thermosensitivity. Increasing extracellular calcium levels eliminates the physiological differences between control and heat shock preparations; excess calcium itself induces thermoprotection at elevated concentrations. These data support the hypothesis that stress-induced neuroprotection at the nerve terminal acts, at least partially, through an alteration to the physiological effects of residual presynaptic calcium.

Enhancement of short-term synaptic plasticity by prior environmental stress.

All chemical synapses can rapidly up- or downregulate the strength of their connections to reshape the postsynaptic signal, thereby stressing the informational importance of specific neural pathways. It is also true that an organism's environment can exert a powerful influence on all aspects of neural circuitry. We investigated the effect of a prior high-temperature stress on the short-term plasticity of a neuromuscular synapse in the hindleg tibial extensor muscle of Locusta migratoria. We found that the prior stress acted to precondition the synapse by increasing the upper temperature limit for synaptic transmission during a subsequent stressful exposure. As well, preexposure to a stressful high-temperature environment increased short-term facilitation of excitatory junction potentials concurrent with a decrease in excitatory junction potential amplitude and a reduction in its temporal parameters. We conclude that a stressful environment can modify synaptic physiological properties resulting in an enhancement of short-term plasticity of the synapse.

Heat-shock-induced thermoprotection of hindleg motor control in the locust.

Functional neuromuscular connections are critical for appropriate behavioural responses, but can be negatively affected by increases in temperature. We investigated the effects of heat shock on the thermosensitivity of a neuromuscular pathway to the hindleg tibial extensor muscle of Locusta migratoria. We found that exposure to heat shock induced thermoprotection of both neuromuscular transmission and extensor muscle contraction by (i) increasing the upper temperature limit for failure, (ii) improving recovery following heat-induced failure and (iii) stabilizing excitatory junction potential amplitude and duration and extensor muscle contraction force at high temperatures. Furthermore, the heat-shock-induced thermoprotection of extensor muscle contraction was not attributable to a protective effect on intrinsic components of muscle contraction. Finally, the use of jumping as a locomotor strategy to avoid capture, a behavioural response dependent upon functionally competent neuromuscular connections at the hindleg tibial extensor muscle, became less sensitive to temperature following heat shock. We conclude that the natural stress response of the locust stabilizes neuromuscular signalling during temperature stress, and that this can underlie a thermoprotection of muscle contraction force and thus alter the thermosensitivity of an escape behaviour critical for survival.

Neuroprotection at Drosophila synapses conferred by prior heat shock.

Synapses are critical sites of information transfer in the nervous system, and it is important that their functionality be maintained under stressful conditions to prevent communication breakdown. Here we show that synaptic transmission at the Drosophila larval neuromuscular junction is protected by prior exposure to heat shock that strongly induces expression of heat shock proteins, in particular hsp70. Using a macropatch electrode to record synaptic activity at individual, visualized boutons, we found that prior heat shock sustains synaptic performance at high test temperatures through pre- and postsynaptic alterations. After heat shock, nerve impulses release more quantal units at high temperatures and exhibit fewer failures of release (presynaptic modification), whereas the amplitude of quantal currents remains more constant than does that in nonheat-shocked controls (postsynaptic modification). The time course of these physiological changes is similar to that of elevated hsp70. Thus, stress-induced neuroprotective mechanisms maintain function at synapses by modifying their properties.