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Decreased expression of hyperpolarisation-activated cyclic nucleotide-gated channel 3 in Hirschsprung's disease

World Journal of Gastroenterology 2015年第18期21卷 5635-5640页

作者： Anne Marie O’Donnell David Coyle prem.puri National Children’s Research Centre Our Lady’s Children’s Hospital Cnmlin Dublin 12 Ireland

AIM: To determine if hyperpolarisation-activated nucleotide-gated(HCN) channels exist in human colon, and to investigate the expression of HCN channels in Hirschsprung's ***:We investigated HCN1,HCN2,HCN3 and HCN4 protein expression in pull-through specimens from patients with Hirschsprung’s disease(HSCR,n=10)using the proximal-most ganglionic segment and distalmost aganglionic segment,as well as in healthy control specimens obtained at the time of sigmoid colostomy closure in children who had undergone anorectoplasty for imperforate anus(n=10).Fluorescent immunohistochemistry was performed to assess protein distribution,which was then visualized using confocal ***:No HCN1 channel expression was observed in any of the tissues *** HCN2 and HCN4proteins were found to be equally expressed in the aganglionic and ganglionic bowel in HSCR and ***3 channel expression was found to be markedly decreased in the aganglionic colon vs ganglionic colon and ***2-4 channels were seen to be expressed within neurons of the myenteric and submucosal plexus of the ganglionic bowel and normal controls,and also co-localised to interstitial cells of Cajal in all tissues ***:We demonstrate HCN channel expression in human colon for the first *** HCN3expression in aganglionic bowel suggests its potential role in HSCR pathophysiology.

关键词： Hyperpolarisation-activated nucleotide-gated Interstitial cells of Cajal Hirschsprung’s Pacemaker Ih current

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Deficiency of platelet-derived growth factor receptor-α-positive cells in Hirschsprung's disease colon

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World Journal of Gastroenterology 2016年第12期22卷 3335-3340页

作者： Anne-Marie O'Donnell David Coyle prem.puri National Children's Research Centre Our Lady's Children's Hospital Crumlin

AIM: To investigate whether the expression of platelet-derived growth factor receptor-α-positive (PDGFRα+)-cells is altered in Hirschsprung’s disease (HD).METHODS: HD tissue specimens (n = 10) were collected at the time of pull-through surgery, while colonic control samples were obtained at the time of colostomy closure in patients with imperforate anus (n = 10). Immunolabelling of PDGFRα+-cells was visualized using confocal microscopy to assess the distribution of these cells, while Western blot analysis was undertaken to quantify PDGFRα protein ***: Confocal microscopy revealed PDGFRα+-cells within the mucosa, myenteric plexus and smooth muscle in normal controls, with a marked reduction in PDGFRα+-cells in the HD specimens. Western blotting revealed high levels of PDGFRα protein expression in normal controls, while there was a striking decrease in PDGFRα protein expression in the HD ***: These findings suggest that the altered distribution of PDGFRα+-cells in both the aganglionic and ganglionic HD bowel may contribute to the motility dysfunction in HD.

关键词： Platelet-derived growth factor receptor alpha Hirschsprung’ s disease Gastrointestinal motility Aganglionosis Myenteric plexus

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Altered tryptophan hydroxylase 2 expression in enteric serotonergic nerves in Hirschsprung's-associated enterocolitis

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World Journal of Gastroenterology 2016年第19期22卷 4662-4672页

作者： David Coyle Justin M Murphy Brian Doyle Anne Marie O’Donnell John Gillick prem.puri Temple Street Children's University Hospital Dublin 1 Ireland National Children's Research Centre Our Lady's Children's Hospital Dublin 12 Ireland

AIM: To determine if expression of colonic tryptophan hydroxylase-2(TPH2), a surrogate marker of neuronal 5-hydroxytryptamine, is altered in Hirschsprung's-associated enterocolitis. METHODS: Entire resected colonic specimens were collected at the time of pull-through operation in children with Hirschsprung's disease(HSCR, n = 12). Five of these patients had a history of pre-operative Hirschsprung's-associated enterocolitis(HAEC). Controls were collected at colostomy closure in children with anorectal malformation(n = 10). The distribution of expression of TPH2 was evaluated using immunofluorescence and confocal microscopy. Protein expression of TPH2 was quantified using western blot analysis in the deep smooth muscle layers. RESULTS: TPH2 was co-expressed in nitrergic and cholinergic ganglia in the myenteric and submucosal plexuses in ganglionic colon in HSCR and healthy controls. Co-expression was also seen in submucosal interstitial cells of Cajal and PDGFRα+ cells. The density of TPH2 immuno-positive fibers decreased incrementally from ganglionic bowel to transition zonebowel to aganglionic bowel in the myenteric plexus. Expression of TPH2 was reduced in ganglionic bowel in those affected by pre-operative HAEC compared to those without HAEC and healthy controls. However, expression of TPH2 was similar or high compared to controls in the colons of children who had undergone diverting colostomy for medically refractory ***: Altered TPH2 expression in colonic serotonergic nerves of patients with HSCR complicated by HAEC may contribute to intestinal secretory and motor disturbances, including recurrent HAEC.

关键词： Serotonin Tryptophan hydroxylase 2 Hirschsprung Enterocolitis Ganglionic

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