Intrinsic neurones and associated cells of the guinea-pig heart in culture

Abstract

This paper describes a method for dissociation of intrinsic neurones from the atria and interatrial septum of newborn guinea-pig heart and their maintenance in culture. The appearance of the cultured intracardiac neurones, muscle and other non-neuronal cell types also present in the preparation has been observed by phase-contrast microscopy. Some of the neurochemical properties of the intracardiac neurones in culture have been investigated using histochemical methods. All the neurones studied were shown to contain acetylcholinesterase. No catecholamine-containing neurones were found. Using an indirect immunofluorescence technique, 20–50% of clearly identifiable neurones in culture contained neuropeptide Y-like immunoreactivity. Vasoactive intestinal polypeptide-like immunoreactive neurones were found in only one out of 15 culture preparations; no substance P-, neurotensin-, or enkephalin-like immunoreactivity was observed. These findings are consistent with those described for intracardiac neurones studied in situ, suggesting that the neurochemical differentiation of the intrinsic heart neurones is retained in culture. The culture preparation provides an opportunity to study the properties and role of intrinsic neurones of the heart. The characteristics of the intracardiac neurones may be distinguished from those of the extrinsic nerve fibres which degenerate in culture. Further, the intracardiac neurones are more accessible to experimental manipulation in culture than in situ.