Cholinergic and nitrergic interneurones in the myenteric plexus of the human colon - PubMed (original) (raw)

Cholinergic and nitrergic interneurones in the myenteric plexus of the human colon

A J Porter et al. Gut. 2002 Jul.

Abstract

Background: Myenteric interneurones are involved in the reflexes that control the motility of the human colon.

Aims: The distribution of choline acetyltransferase (ChAT) and nitric oxide synthase (NOS) immunoreactivity in myenteric interneurones was investigated in this study.

Methods: DiI (1,1'- didodecyl 3,3,3',3'-indocarbocyanine perchlorate) was applied to the myenteric plexus of the human colon followed by organotypic culture. Retrogradely labelled neurones, with projections longer than motor neurones (>10 mm), were studied to exclude motor neurone populations. ChAT and NOS immunoreactivity was then determined in the interneurones.

Results: We found that 90% of interneurones projecting orally contained ChAT and none contained NOS. Ninety five per cent of descending interneurones were labelled with ChAT and/or NOS antisera; 46% contained NOS immunoreactivity alone, 20% contained ChAT immunoreactivity alone, and 29% contained both ChAT and NOS. Anally directed interneurones had significantly longer projections than orally projecting interneurones.

Conclusions: Nearly all interneurones contain either NOS or ChAT immunoreactivity. Orally projecting interneurones are of two types: 90% contain ChAT alone and the remainder contain immunoreactivity for neither ChAT nor NOS. There are three main types of anally projecting interneurones: the largest, which contains NOS but not ChAT, and the two smaller classes which contain ChAT and NOS, and CHAT alone.

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Figures

Figure 1

Figure 1

Scatterplot showing the location of 1,1`- didodecyl 3,3,3`,3`-indocarbocyanine perchlorate (DiI) containing neurones with choline acetyltransferase (ChAT) or nitric oxide synthase (NOS) immunoreactivity in a single preparation. The shaded strip indicates the DiI application site. Almost all neurones with ascending projections contain ChAT whereas there are several subgroups with descending projections. Calibration bar, 10 mm.

Figure 2

Figure 2

Myenteric neurone labelled 16 mm anal to the 1,1`- didodecyl 3,3,3`,3`-indocarbocyanine perchlorate (DiI) application site on the myenteric plexus. The nerve cell body was immunoreactive for choline acetyltransferase (ChAT) but not nitric oxide synthase (NOS) (filled arrows). A neurone not labelled with DiI (open arrow) was immunoreactive for both ChAT and NOS. Calibration bar, 20 μm.

Figure 3

Figure 3

Histograms showing the distribution of (A) choline acetyltransferase (ChAT) +/nitric oxide synthase (NOS)−, (B) ChAT−/NOS+, (C) ChAT+/NOS+, and (D) ChAT−/NOS− neurones as a proportion of the total number of 1,1`- didodecyl 3,3,3`,3`-indocarbocyanine perchlorate (DiI) labelled neurone. Data were pooled from the three preparations labelled with both ChAT and NOS antisera. Dotted lines at −10/+10 indicate the projection limit of non-interneurones; hence neurones outside these limits are interneurones.

Figure 4

Figure 4

Anally projecting interneurone labelled 11 mm from the 1,1`- didodecyl 3,3,3`,3`-indocarbocyanine perchlorate (DiI) application site. The nerve cell body was immunoreactive for nitric oxide synthase (NOS) but not choline acetyltransferase (ChAT) (filled arrows). A neurone not labelled with DiI (open arrow) was immunoreactive for both ChAT and NOS. Calibration bar, 20 μm.

Figure 5

Figure 5

Descending interneurone containing both choline acetyltransferase (ChAT) and nitric oxide synthase (NOS) (arrows). It was located 20 mm oral to the 1,1`- didodecyl 3,3,3`,3`-indocarbocyanine perchlorate (DiI) application site. Calibration bar, 20 μm.

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