Cuprolinic Blue (quinolinic phthalocyanine) counterstaining of enteric neurons for peroxidase immunocytochemistry

Holst, M.-C.; Powley, Terry L.

doi:10.1016/0165-0270(95)00064-x

Cited by 57 publications

(49 citation statements)

References 26 publications

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“…Neurons were counted in fetal gut that was immunostained with the neuronal marker PGP9.5 (protein gene product; a neuronal form of ubiquitin hydrolase) (supplemental Tables 1, 2, available at www.jneurosci.org). To count the total number of neurons in postnatal bowel, the laminar preparations were stained for 1 hr at 37°C with cuprolinic blue, which is neuron selective and reveals the entire neuronal population (0.5% in 0.05 mM sodium acetate buffer, pH 5.6, containing 1.0 M MgCl 2 ) (Heinicke et al, 1987;Holst and Powley, 1995;Karaosmanoglu et al, 1996). Nerve cell bodies were counted in 10 contiguous nonoverlapping rectangular fields covering 1.254 mm 2 .…”

Section: Methodsmentioning

confidence: 99%

Bone Morphogenetic Protein-2 and -4 Limit the Number of Enteric Neurons But Promote Development of a TrkC-Expressing Neurotrophin-3-Dependent Subset

Chalazonitis¹,

D’Autréaux²,

Guha³

et al. 2004

J. Neurosci.

121

212

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The hypothesis that BMPs (bone morphogenetic proteins), which act early in gut morphogenesis, also regulate specification and differentiation in the developing enteric nervous system (ENS) was tested. Expression of BMP-2 and BMP-4, BMPR-IA (BMP receptor subunit), BMPR-IB, and BMPR-II, and the BMP antagonists, noggin, gremlin, chordin, and follistatin was found when neurons first appear in the primordial bowel at embryonic day 12 (E12). Agonists, receptors, and antagonists were detected in separated populations of neural crestand noncrest-derived cells. When applied to immunopurified E12 ENS precursors, BMP-2 and BMP-4 induced nuclear translocation of phosphorylated Smad-1 (Sma and Mad-related protein). The number of neurons developing from these cells was increased by low concentrations and decreased by high concentrations of BMP-2 or BMP-4. BMPs induced the precocious appearance of TrkC-expressing neurons and their dependence on neurotrophin-3 for survival. BMP-4 interacted with glial cell line-derived neurotrophic factor (GDNF) to enhance neuronal development but limited GDNF-driven expansion of the precursor pool. BMPs also promoted development of smooth muscle from mesenchymal cells immunopurified at E12. To determine the physiological significance of these observations, the BMP antagonist noggin was overexpressed in the developing ENS of transgenic mice under the control of the neuron-specific enolase promoter. Neuronal numbers in both enteric plexuses and smooth muscle were increased throughout the postnatal small intestine. These increases were already apparent by E18. In contrast, TrkC-expressing neurons decreased in both plexuses of postnatal nogginoverexpressing animals, again an effect detectable at E18. BMP-2 and/or BMP-4 thus limit the size of the ENS but promote the development of specific subsets of enteric neurons, including those that express TrkC.

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Section: Methodsmentioning

confidence: 99%

Bone Morphogenetic Protein-2 and -4 Limit the Number of Enteric Neurons But Promote Development of a TrkC-Expressing Neurotrophin-3-Dependent Subset

Chalazonitis¹,

D’Autréaux²,

Guha³

et al. 2004

J. Neurosci.

121

212

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“…Cuprolinic-blue histochemistry provides selective and complete staining of enteric neurons (Heinicke et al 1987;Holst and Powley 1995;Karaosmanoglu et al 1996;Phillips et al 2004). We used wholemounts of the muscularis propria that were prepared previously for quantification only .…”

Section: Distribution Of Myenteric Ganglia In Esophageal Wholemountsmentioning

confidence: 99%

Number and Distribution of Intraganglionic Laminar Endings in the Mouse Esophagus as Demonstrated with Two Different Immunohistochemical Markers

Raab

Neuhuber

2005

J Histochem Cytochem.

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S U M M A R YIntraganglionic laminar endings (IGLEs) represent the only vagal mechanosensory terminals in the tunica muscularis of the esophagus. Two specific markers for IGLEs were recently described in mouse: the purinergic P2 ϫ 2 receptor and the vesicular glutamate transporter 2 (VGLUT2). This study aimed at comparing both markers with respect to their suitability for quantitative analysis. We counted IGLEs immunostained for VGLUT2 and P2 ϫ 2 , respectively, and mapped their distribution in esophageal wholemounts of C57Bl/6 mice. Numbers and distribution of IGLEs were compared with those of myenteric ganglia as demonstrated by cuprolinic blue histochemistry. Whereas the distribution of VGLUT2-immunopositive IGLEs closely matched that of myenteric ganglia, P2 ϫ 2 -immunopositive IGLEs were rarely found in upper and middle esophagus but increasingly in its lower parts. P2 ϫ 2 stained only half the number of IGLEs found with VGLUT2 immunostaining. We also investigated the correlation between anterograde tracing and immunohistochemistry for identifying IGLEs. Confocal microscopy revealed colocalization of all three markers in ‫ف‬ 50% of IGLEs. The remaining IGLEs showed only tracer and VGLUT2 labeling but no P2 ϫ 2 immunoreactivity. Thus, VGLUT2 and P2 ϫ 2 represent two specific markers for qualitative demonstration of esophageal IGLEs. However, VGLUT2 may be superior to P2 ϫ 2 as a quantitative marker for IGLEs in the esophagus of C57Bl/6 mice.

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“…The protocol used was based on the protocol described by Holst and Powley (1995), but modified in our laboratory (Van Ginneken et al in press).…”

Section: Cuprolinic Blue Stainingmentioning

confidence: 99%

“…Cuprolinic blue, an analogue of the well-known phthalocyanin dye Alcian Blue 8GX, has received renewed interest as a marker for the entire enteric neuron population (Scott 1972;Heinicke et al 1987;Holst and Powley 1995;Karaosmanoglu et al 1996). In combination with high salt concentrations, it has a remarkable specificity for single-stranded RNA.…”

mentioning

confidence: 99%

“…One of its advantages is that, under these specific conditions, cuprolinic blue does not bind to nuclear DNA or other compounds commonly found in tissues (Scott 1972;Mendelson et al 1983;Tas et al 1983). Consequently, some investigators have concluded that, given the high concentration of RNA in neurons, enteric neurons are the only structures that can be stained in gastrointestinal whole-mount preparations (Heinicke et al 1987;Holst and Powley 1995;Karaosmanoglu et al 1996). The aim of this study was to elaborate different combinations between cuprolinic blue staining and peroxidase immunohistochemistry and/or enzyme histochemistry to provide a readily usable procedure that simultaneously visualizes the total population as well as subsets of enteric neurons.…”

mentioning

confidence: 99%

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Microwave Staining of Enteric Neurons Using Cuprolinic Blue (Quinolinic Phthalocyanin) Combined with Enzyme Histochemistry and Peroxidase Immunohistochemistry

Ginneken

Smet

Meir

et al. 1999

J Histochem Cytochem.

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SUMMARYMethods that visualize subsets as well as the entire enteric neuron population are not readily available or have proved to be unreliable. Therefore, we attempted to combine NADPH-d histochemistry, AChE histochemistry, and CGRP immunohistochemistry, techniques that mark subsets of enteric neurons, with a technique that appeared to visualize the entire enteric neuron population, the cuprolinic blue staining method. To guarantee representative staining results, the individual staining methods were modified by using microwaves. In addition, this preserved the characteristics of each of the individual techniques. The distribution of NADPH-d, AChE, and CGRP corresponded well with previous morphological and physiological reports. Consequently, the different combinations gave rise to rapid, useful, and ready-to-use double labeling techniques. Their main advantage is that they simultaneously visualize the total population as well as subsets of enteric neurons. (J Histochem Cytochem 47:13-21, 1999)

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Cuprolinic Blue (quinolinic phthalocyanine) counterstaining of enteric neurons for peroxidase immunocytochemistry

Cited by 57 publications

References 26 publications

Bone Morphogenetic Protein-2 and -4 Limit the Number of Enteric Neurons But Promote Development of a TrkC-Expressing Neurotrophin-3-Dependent Subset

Bone Morphogenetic Protein-2 and -4 Limit the Number of Enteric Neurons But Promote Development of a TrkC-Expressing Neurotrophin-3-Dependent Subset

Number and Distribution of Intraganglionic Laminar Endings in the Mouse Esophagus as Demonstrated with Two Different Immunohistochemical Markers

Microwave Staining of Enteric Neurons Using Cuprolinic Blue (Quinolinic Phthalocyanin) Combined with Enzyme Histochemistry and Peroxidase Immunohistochemistry

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