NDLI: Correlation of Enteric NADPH-d Positive Cell Counts with the Duration of Incubation Period in NADPH-d Histochemistry

Content Provider	Springer Nature Link
Author	Cserni, Tamas O’ Donnel, Annemarie Paran, Sri Puri, Prem
Copyright Year	2008
Abstract	Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) staining can be used in the enteric nervous system to determine nitrergic neuronal counts, critical in motility disorders such as intestinal neuronal dysplasia and hypoganglionosis. The reported incubation periods of specimens with NADPH-d staining solution has varied from 2 to 24 h. The aim of this study is to investigate the impact of the incubation period on the overall NADPH-d positive cell counts in porcine rectal submucosal plexus. The submucosal plexus of rectal specimens from 12-week-old pigs (n = 5) were studied. Conventional frozen sections were used to identify nitrergic neurons while whole-mount preparations were used to quantify the effect of prolonged duration of incubation on positively identified ganglion cells with NADPH-d histochemistry. The same submucosal ganglia on the conventional sections, and a minimum of 12 ganglia per whole-mount preparation specimen were photographed sequentially at 2, 6, and 24 h and used to count the number of nitrergic cells per ganglion. The same staining solution was used throughout the experiment. Results were analysed using a one-way ANOVA test. Prolonged incubation with the staining solution revealed new NADPH-d positive cells in the ganglia on the conventional sections. The total number of neurons counted in the 12 adjacent ganglia in the whole-mount specimens was 180 ± 55, the mean neuronal cell per ganglion was 15 ± 8 after 2 h of incubation. This increased to 357 ± 17, and to 29 ± 12 after 6 h (p < 0.05). A further increase was observed of 515 ± 19 and 43 ± 17 after 24 h (p < 0.05). When the photomicrographs were retrospectively analysed, not even the outline of the neuronal cells that stained with prolonged incubation was evident at the earlier time points. NADPH-d positive cell counts increase in proportion to the duration of incubation in NADPH-d histochemistry. Comparative studies attempting to quantify nitrergic cell counts in dysmotility disorders must take into account the variability in NADPH-d positive cell count associated with prolonged incubation in NADPH-d histochemistry.
Starting Page	103
Ending Page	107
Page Count	5
File Format	PDF
ISSN	12194956
Journal	Pathology & Oncology Research
Volume Number	15
Issue Number	1
e-ISSN	15322807
Language	English
Publisher	Springer Netherlands
Publisher Date	2008-07-10
Publisher Place	Dordrecht
Access Restriction	Subscribed
Subject Keyword	Positive cell counts Incubation time NADPH-d histochemistry Biomedicine general Immunology Pathology Oncology Cancer Research
Content Type	Text
Resource Type	Article
Subject	Cancer Research Pathology and Forensic Medicine Oncology

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