Central control of atrio-ventricular conduction and left ventricular contractility in the cat heart: Synaptic interactions of vagal preganglionic neurons in the nucleus ambiguus with neuropeptide Y-immunoreactive nerve terminals.

In the cat, vagal postganglionic controls of heart rate, atrio-ventricular (AV) conduction and left ventricular contractility are mediated by three separate intrinsic cardiac ganglia, the sinoatrial (SA), AV and cranioventricular (CV) ganglia, respectively. The vagal preganglionic neurons (VPNs) that project to these ganglia are located in the ventrolateral nucleus ambiguus (NA-VL). We have previously shown that the VPNs projecting to the SA, AV and CV ganglia are distinct from one another. We have also demonstrated that neuropeptide Y-immunoreactive (NPY-IR) axon terminals synapse upon VPNs projecting to the SA ganglion. In the present study, we test the hypothesis that those VPNs projecting to the AV ganglion (negative dromotropic VPNs) and those projecting to the CV ganglion (negative inotropic VPNs) are innervated by NPY-IR terminals in NA-VL. A retrograde tracer was injected into the AV or CV ganglion of the cat, and the brains subsequently processed for visualization of tracer and the immunocytochemical visualization of NPY by dual labeling electron-microscopic methods. We observed that 11+/-5% of all axodendritic synapses and 8+/-6% of all axosomatic synapses upon negative inotropic VPNs were NPY-IR. Furthermore, 19+/-14% of all axodendritic synapses upon negative dromotropic VPNs were NPY-IR. A few NPY-IR axosomatic synapses upon negative dromotropic neurons were also observed. NPY-IR terminals in NA-VL occasionally formed axosomatic synapses with NPY-IR neurons and axoaxonic synapses with unlabeled terminals. These results suggest that central NPY afferents to the NA-VL modulate the vagal preganglionic control of AV conduction and left ventricular contractility.

Intercellular interactions in the mammalian olfactory nerve.

The small, unmyelinated axons of olfactory sensory neurons project to the olfactory bulb in densely packed fascicles, an arrangement conducive to axo-axonal interactions. We recently demonstrated ephaptic interactions between these axons in the olfactory nerve layer, the layer of the olfactory bulb in which the axon fascicles interweave and rearrange extensively. In the present study, we hypothesized that the axons, which express connexins, may have another mode of communication: gap junctions. Previous transmission electron microscopy (TEM) studies have failed to demonstrate such junctions. However, the definitive method for detecting gap junctions, freeze fracture, has not been used to examine the interaxonal connections of the olfactory nerve layer. Here, we apply a combined approach of TEM and freeze fracture to determine if gap junctions are present between the olfactory axons. Gap junctions involving olfactory axons were not found. However, by freeze fracture, P faces of both the axons and ensheathing cells (glia that surround the axon fascicles) contained distinctive linear arrays of particles, aligned along the small columns of extracellular space. In axons, few intramembranous particles were present outside of these arrays. Multi-helix proteins, including ion channels and connexin hemichannels, have been shown to be visible as particles by freeze fracture. This suggests that the proteins important for signal transmission are confined to the linear arrays. Such an arrangement would facilitate ephaptic transmission, calcium waves, current oscillations, and paracrine communication and may be important for olfactory neural code processing.