We set out to determine the projections of the major immunohistochemically-defined populations of intrinsic cardiac neurons to different target tissues within the guinea-pig heart. Ultrastructural studies, and immunoreactivity to the neuronal marker, neuron-specific enolase, suggested that the number of axons of intrinsic neurons in most regions of the heart was low when compared with the populations of axons projecting from extrinsic sensory and sympathetic ganglia. Multiple-labelling immunofluorescence was used to demonstrate the terminals of the major populations of peptide-containing intrinsic neurons. The intrinsic nature of peptide-containing axons was confirmed by long-term organotypic culture of cardiac tissue, which resulted in degeneration of axons of extrinsic neurons. The relative density and peptide content of intrinsic axons throughout the heart was not consistent with the relative proportions of peptide-containing intracardiac nerve cell bodies observed previously. The most commonly-encountered axons contained immunoreactivity (IR) to vasoactive intestinal peptide (VIP) alone, although nerve cell bodies with VIP constituted less than 5% of the total population of intrinsic neurons. Populations of axons containing IR to somatostatin alone, somatostatin and substance P, neuropeptide Y (NPY) alone, somatostatin and NPY, or VIP and NPY, also were observed. Intrinsic axons containing substance P-IR were very rare, much more so than would be predicted from the peptide content of intrinsic nerve cell bodies. The regions of the heart with the most dense innervation by axons of intrinsic neurons were the cardiac valves, the atrio-ventricular node and the sino-atrial node. Each of these targets was innervated by several populations of peptide-containing axons. Thus, each population of peptide-containing intrinsic neurons projected to a variety of target tissues within the heart. One possible interpretation of these results is that immunohistochemically-distinct populations of intrinsic neurons belong to different functional classes of neurons (sensory neurons, interneurons, final motor neurons), each of which innervates many regions of the heart.
- Cardiac valve
- Intracardiac neuron
- Neuropeptide Y
- Parasympathetic neuron
- Vasoactive intestinal peptide