Serotonin (5-HT) powerfully affects small intestinal motility and 5-HT-immunoreactive (IR) neurones are highly conserved between species. 5-HT synthesis in central neurones and gastrointestinal mucosa depends on tissue-specific isoforms of the enzyme tryptophan hydroxylase (TPH). RT-PCR identified strain-specific expression of a polymorphism (1473C/G) of the tph2 gene in longitudinal muscle-myenteric plexus preparations of C57Bl/6 and Balb/c mice. The former expressed the high-activity C allele, the latter the low-activity G allele. Confocal microscopy was used to examine close contacts between 5-HT-IR varicosities and myenteric neurones immunoreactive for neuronal nitric oxide synthase (NOS) or calretinin in these two strains. Significantly more close contacts were identified to NOS-(P < 0.05) and calretinin-IR (P < 0.01) neurones in C57Bl/6 jejunum (NOS 1.6 ± 0.3, n = 52; calretinin 5.2 ± 0.4, n = 54), than Balb/c jejunum (NOS 0.9 ± 0.2, n = 78; calretinin 3.5 ± 0.3, n = 98). Propagating contractile complexes (PCCs) were identified in the isolated jejunum by constructing spatiotemporal maps from video recordings of cannulated segments in vitro. These clusters of contractions usually arose towards the anal end and propagated orally. Regular PCCs were initiated at intraluminal pressures of 6 cmH 2 O, and abolished by tetrodotoxin (1 μm). Jejunal PCCs from C57Bl/6 mice were suppressed by a combination of granisetron (1 μm, 5-HT 3 antagonist) and SB207266 (10 nm, 5-HT 4 antagonist), but PCCs from Balb/c mice were unaffected. There were, however, no strain-specific differences in sensitivity of longitudinal muscle contractions to exogenous 5-HT or blockade of 5-HT 3 and 5-HT 4 receptors. These data associate a genetic difference with significant structural and functional consequences for enteric neural serotonergic pathways in the jejunum.
Polarized outputs of myenteric interneurons in guinea-pig small intestine have been well studied. However, the variety of motility patterns exhibited suggests that some interneuron targets remain unknown. We used antisera selected to distinguish interneuron varicosities and known myenteric neuron types to investigate outputs of three interneuron classes in guinea-pig jejunum; two classes of descending interneurons immunoreactive (IR) for somatostatin (SOM) or nitric oxide synthase (NOS)/vasoactive intestinal peptide (VIP), and one class of ascending interneurons [calretinin/enkephalin (ENK)-IR]. Varicosities apposed to immunohistochemically identified cell bodies were quantified by confocal microscopy. Intrinsic sensory neurons (calbindin-IR) were apposed by few varicosities. Cholinergic secretomotor neurons (neuropeptide Y-IR) were apposed by many SOM-IR varicosities. Longitudinal muscle excitatory motor neurons (calretinin-IR) were apposed by some VIP- and ENK-IR varicosities, but few SOM-IR varicosities. Ascending interneurons (calretinin-IR) were apposed by many varicosities of all types. NOS-IR interneurons and inhibitory motor neurons were apposed by numerous VIP-IR and SOM-IR varicosities. NOS-IR short inhibitory motor neurons were apposed by significantly fewer ENK-IR varicosities than other NOS-IR neurons. Based on the specific chemical coding of ascending (ENK) and descending (SOM) interneurons, we conclude that cholinergic secretomotor neurons and short inhibitory neurons are located in descending reflex pathways, while ascending interneurons and NOS-IR descending interneurons are focal points at which ascending and descending pathways converge.
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