Quantitative evaluation of myenteric ganglion cells in normal human left colon: implications for histopathological analysis

Abstract

The analysis of myenteric neurons is becoming increasingly important for the assessment of enteric nervous system injury and degeneration occurring in motor disorders of the gut. Limited information is presently available on the quantitative estimation of myenteric neurons and glial cells in paraffin-embedded colonic sections; additional data would be useful for diagnostic purposes. In this morphometric study, we performed immunohistochemistry to count myenteric neurons and glial cells in paraffin sections of human colon. Serial cross sections of formalin-fixed paraffin-embedded full-thickness normal human left colon (n = 10, age-range: 50-72 years) were examined. HuC/D and S100beta antigens were found to be the best markers for the detection of neurons and glial cells, respectively. Significant correlations were noted between the numbers of neurons/glial cells and the respective myenteric ganglion areas. These findings suggest that HuC/D-S100beta-immunostained paraffin cross sections of human colon can be regarded as valuable tools for the quantitative estimation of myenteric neurons and glial cells. Based on the present method, only a limited number of paraffin sections are needed for reliable quantitative assessments of myenteric ganglion cells, thus allowing fast and simple approaches in the settings of the histopathological diagnosis of colonic motility disorders and retrospective evaluations of pathological archival tissue specimens.

Parkinson’s disease is not associated with gastrointestinal myenteric ganglion neuron loss

“…The quantitative method used in this study was described recently to provide accurate detection and reliable quantification of immunolabeled myenteric ganglion cells in human colon [26,10,57]. There was very close agreement between our results and those of Ippolito et al (2009) indicating this method is valid, reproducible, and provides a reliable indication of myenteric neuron density in the colon [26]. Our results also indicate it is generalizable to other segments of the GI tract.…”

“…Several studies have been performed on paraffin sections to evaluate the organization of the myenteric plexus [9,5,24] and to obtain relative proportions of myenteric neurochemical phenotypes in humans and other mammals [3,38]. The quantitative method used in this study was described recently to provide accurate detection and reliable quantification of immunolabeled myenteric ganglion cells in human colon [26,10,57]. There was very close agreement between our results and those of Ippolito et al (2009) indicating this method is valid, reproducible, and provides a reliable indication of myenteric neuron density in the colon [26].…”

“…For each case, neuronal profiles and ganglion area were both summed within segments. The number of neurons per ganglion area (mm 2 ) was calculated for each case as Σ neuron profiles ÷ Σ area measurements [26]. …”

Vascular Generation of Tumor Necrosis Factor-α Reduces Nitric Oxide Availability in Small Arteries From Visceral Fat of Obese Patients

“…Tissue specimens were immediately fixed in cold neutral 4% formaldehyde and then paraffin-embedded. Eight-micrometer-thick sections were microwaved; immunostained by rabbit anti-TNF-␣ polyclonal antibodies (Abcam, Cambridge, Massachusetts); and treated with biotinylated anti-rabbit immunoglobulins, peroxidase-labeled streptavidin complex (Vector Laboratories, Burlingame, California), and 3,3 diaminobenzidine tetra-hydrochloride (DAB) (Dakopatts, Glostrup, Denmark), as previously reported (19). Negative controls were obtained by substituting the primary antibody with preimmune rabbit serum.…”

Neurochemical phenotypes of myenteric neurons in the rhesus monkey

“…ENS neurons have been classified in numerous ways including shape, anatomical projections, electrophysiological properties, function, and neurotransmitter content (Bornstein et al, 2004; Furness, 2006; Schemann, 2005; Costa and Brookes, 2008). The neurochemical phenotype of enteric neurons is critical to their function and has been extensively studied in guinea pigs, mice, rats, and humans (Gabella, 1987; Ekblad et al, 1988, 1994a; Costa et al, 1992; Wattchow et al, 1997, 2008; Porter et al, 1997; 2002; Lomax and Furness, 2000; Furness, 2000, 2006; Anlauf et al, 2003; Neunlist et al, 2003; Pimont et al, 2003; Bornstein et al, 2004; Ganns et al, 2006; Murphy et al, 2007; Qu et al, 2008; Ippolito et al, 2009; Mongardi Fantaguzzi et al, 2009). While essentially every classical neurotransmitter and neuropeptide is expressed in the ENS, particularly prominent roles in mammalian enteric neurotransmission have been ascribed to nitric oxide (NO), vasoactive intestinal peptide (VIP), catecholamines, and acetylcholine (Ach).…”