Increased expression of nitric oxide synthase in cultured neurons from adult rat colonic submucous ganglia

Neuronal plasticity in the enteric nervous system (ENS) is probably a key step in intestinal adaptation during growth, maturation and ageing as well as in several pathophysiological situations. Studies on cultured myenteric neurons have revealed an increased vasoactive intestinal peptide (VIP) expression in neuronal nitric oxide synthase (NOS)-expressing neurons. In addition, both VIP and nitric oxide (NO) promote survival of cultured myenteric neurons. The aim of the present study was to investigate possible changes in the expression of VIP and NOS in cultured submucous neurons from adult rat large intestine. Submucous neurons were cultured as explants or as dissociated neurons for 3 and 8 days. Immunocytochemistry was used to determine the proportions of neurons containing VIP or NOS in preparations of uncultured controls (reflects the conditions in vivo) and in cultured explants of submucosa and dissociated submucous neurons. In situ hybridization was used to determine changes in the expressions of NOS and VIP mRNA. The relative number of NOS-expressing neurons increased significantly during culturing. The percentage of all neurons expressing NOS was 22% in controls, while approximately 50% of the cultured submucous neurons expressed NOS. VIP-expressing neurons constituted approximately 80% of all submucous neurons in controls as well as in cultured explants or dissociated neurons. Studies on coexistence revealed that the VIP-containing neurons were the ones that started to express NOS during culture. The induced expression of NOS in cultured adult submucous neurons indicates that nitric oxide, possibly in cooperation with VIP, is important for neuronal adaptation, maintenance and survival.