Multiple sites of action potential initiation increase neuronal firing rate.

Sensory input to an individual interneuron or motoneuron typically evokes activity at a single site, the initial segment, so that firing rate reflects the balance of excitation and inhibition there. In a network of cells that are electrically coupled, a sensory input produced by appropriate, localized stimulation can cause impulses to be initiated in several places. An example in the leech is the chain of S cells, which are critical for sensitization of reflex responses to mechanosensory stimulation. S cells, one per segment, form an electrically coupled chain extending the entire length of the CNS. Each S cell receives input from mechanosensory neurons in that segment. Because impulses can arise in any S cell and can reliably propagate throughout the chain, all the S cells behave like a single neuron with multiple initiation sites. In the present experiments, well-defined stimuli applied to a small area of skin evoked mechanosensory action potentials that propagated centrally to several segments, producing S cell impulses in those segments. Following pressure to the skin, impulses arose first in the S cell of the same segment as the stimulus, followed by impulses in S cells in other segments. Often four or five separate initiation sites were observed. This timing of impulse initiation played an important role in increasing the frequency of firing. Impulses arising at different sites did not usually collide but added to the total firing rate of the chain. A computational model is presented to illustrate how mechanosensory neurons distribute the effects of a single sensory stimulus into spatially and temporally separated synaptic input. The model predicts that changes in impulse propagation in mechanosensory neurons can alter S cell frequency of firing by changing the number of initiation sites.

Action potential reflection and failure at axon branch points cause stepwise changes in EPSPs in a neuron essential for learning.

In leech mechanosensory neurons, action potentials reverse direction, or reflect, at central branch points. This process enhances synaptic transmission from individual axon branches by rapidly activating synapses twice, thereby producing facilitation. At the same branch points action potentials may fail to propagate, which can reduce transmission. It is now shown that presynaptic action potential reflection and failure under physiological conditions influence transmission to the same postsynaptic neuron, the S cell. The S cell is an interneuron essential for a form of nonassociative learning, sensitization of the whole body shortening reflex. The P to S synapse has components that appear monosynaptic (termed "direct") and polysynaptic, both with glutamatergic pharmacology. Reflection at P cell branch points on average doubled transmission to the S cell, whereas action potential failure, or conduction block, at the same branch points decreased it by one-half. Each of two different branch points affected transmission, indicating that the P to S connection is spatially distributed around these branch points. This was confirmed by examining the locations of individual contacts made by the P cell with the S cell and its electrically coupled partner C cells. These results show that presynaptic neuronal morphology produces a range of transmission states at a set of synapses onto a neuron necessary for a form of learning. Reflection and conduction block are activity-dependent and are basic properties of action potential propagation that have been seen in other systems, including axons and dendrites in the mammalian brain. Individual branch points and the distribution of synapses around those branch points can substantially influence neuronal transmission and plasticity.

Synaptic facilitation by reflected action potentials: enhancement of transmission when nerve impulses reverse direction at axon branch points.

A rapid, reversible enhancement of synaptic transmission from a sensory neuron is reported and explained by impulses that reverse direction, or reflect, at axon branch points. In leech mechanosensory neurons, where one can detect reflection because it is possible simultaneously to study electrical activity in separate branches, action potentials reflected from branch points within the central nervous system under physiological conditions. Synapses adjacent to these branch points were activated twice in rapid succession, first by an impulse arriving from the periphery and then by its reflection. This fast double-firing facilitated synaptic transmission, increasing it to more than twice its normal level. Reflection occurred within a range of resting membrane potentials, and electrical activity produced by mechanical stimulation changed membrane potential so as to produce and cease reflection. A compartmental model was used to investigate how branch-point morphology and electrical activity contribute to produce reflection. The model shows that mechanisms that hyperpolarize the membrane so as to impair action potential propagation can increase the range of structures that can produce reflection. This suggests that reflection is more likely to occur in other structures where impulses fail, such as in axons and dendrites in the mammalian brain. In leech sensory neurons, reflection increased transmission from central synapses only in those axon branches that innervate the edges of the receptive field in the skin, thereby sharpening spatial contrast. Reflection thus allows a neuron to amplify synaptic transmission from a selected group of its branches in a way that can be regulated by electrical activity.

Differential regulation of synaptic vesicle protein genes by target and synaptic activity.

Differentiation of presynaptic nerve terminals involves changes in gene expression; these may be regulated by synaptic transmission and/or by contact with the target muscle. To gain insight into the control of presynaptic differentiation, we examined the regulation by target and synaptic activity of synaptic vesicle protein (SVP) genes in the chick ciliary ganglion (CG). In the CG, two SVP genes, synaptotagmin I (syt I) and synaptophysin II (syp II), are coordinately upregulated at the time of target contact. To test the hypothesis that this upregulation is induced by target contact, we examined mRNA levels of syt I and syp II in CGs from embryos in which one eye had been removed before axon outgrowth. As expected, target removal prevented the normal upregulation of syt I mRNA in the deprived ganglion. In contrast, and unexpectedly, syp II mRNA upregulation was not affected. The target dependence of syt I upregulation was not attributable to nerve-muscle transmission, because blockade of this transmission had no effect on SVP mRNA levels. Surprisingly, blockade of synapses onto CG neurons from the brain also did not affect syt I mRNA levels but increased levels of syp II mRNA. We conclude that contact with target induces upregulation of syt I mRNA, which is the case for spinal motor neurons. However, the normal upregulation of syp II mRNA is not controlled by the same signal(s). Instead, our results suggest that these two SVP genes are differentially regulated, both by target contact and by blockade of synaptic transmission.