Expression of choline acetyltransferase immunoreactivity in guinea pig cardiac ganglia

Recent reports indicate that a considerable amount of heterogeneity exists amongst cardiac postganglionic neurons in their chemical coding patterns and electrical properties, and that some of these cells may serve in roles as sensory and interganglionic neurons as well as motor neurons. This study was undertaken to ascertain whether or not all of these neurons are cholinergic by immunostaining whole-mount preparations of the guinea pig heart for choline acetyltransferase (ChAT). Counts of neurons that were immunostained for microtubule-associated protein-2 revealed that about 1000 neurons exist in about 100 ganglia on the posterior atrial surface. ChAT immunoreactivity was expressed by all of the postganglionic neurons in the cardiac ganglia, including the 5% of neurons that also expressed immunoreactivity for nitric oxide synthase. Varicose nerve fibers that were immunoreactive for ChAT were abundant in ganglia, with every cardiac neuron lying in close apposition to one or more labelled varicosities. ChAT-immunoreactive nerve fibers were also observed in large vagosympathetic fiber bundles, in interganglionic fiber bundles, and passing individually within the myocardium. Immunoreactivity for ChAT was also observed in a large proportion of the small tyrosine hydroxylase-immunoreactive neurons that exist in guinea pig cardiac ganglia. These results indicate that all postganglionic neurons in guinea pig cardiac ganglia are likely to utilize acetylcholine as a neurotransmitter, regardless of their functional role in circuitry of cardiac innervation, and each of these neurons is likely to receive cholinergic input.