Aren’t the enteric neurons cholinergic?

They are cholinergic which means ChAT and nNOS antibodies could be an option instead? If so, why to choose ChAT /nNOS combination instead of HuC/HuD or PGP 9.5? I have lack of practical experience in enteric neuroscience area so it would be really helpful if you could explain to question of why.

Thanks in advance

The combination of antibodies for HuD/C and GFAP seems to work well for mouse enteric nervous system.

See: Cell Mol Gastroenterol Hepatol. 2018 May 29;6(3):321-344.

Thank you Menachem Hanani, this article gonna be helpful for sure.

It also really depends on what you want to see. If you are more interested in nerve fibers and neuron processes you might want to try PGP9.5 or have a look at beta III tubulin, which is neuron specific as well. This antibody is mainly used in enteric neuroscience, as well as HuC/D. HuC/D is very useful if you want to image neuronal cell bodies. ChAT and nNOS antibodies can be used if you want to show that different neuronal subtypes are present in the ENS. Then you could also stain for calbindin, VIP, calretinin, etc. But for a general overview of neurons and glial cells I would use HuC/D or tub bIII, and S100b or GFAP (try different combinations to see what works best for you). Sox10 will show you both neuronal precursors as well as (mature) enteric glial cells. Hope this helps!

Thank you Simone Schonkeren, explains all the questions in my mind. And it will be helpful for my work, no doubt!

In response to a previous comment: Many enteric neurons are indeed cholinergic, but not all of them are cholinergic (see for example Anat Rec. 1998;251:185-99). Therefore, labeling cholinergic neurons is not suitable for staining the entire population of enteric neurons.