If you're using NeuN as a marker you should be aware of its nature and limitations.

Firstly, it's actually the DNA splicing regulator Fox-3 (Kim, Adelstein & Kawamoto, J Biol Chem. 2009;284(45):31052-61. doi: 10.1074/jbc.M109.052969) and NeuN antibodies cross-react with the synaptic protein synapsin I.

Secondly, NeuN is specific to neurons, but not all neurons express NeuN; specifically, it is not expressed by cerebellar Purkinje neurons, olfactory bulb mitral cells (Mullen, Buck & Smith, Development. 1992;116(1):201-11. http://www.ncbi.nlm.nih.gov/pubmed/1483388), sympathetic chain ganglion neurons, cortical Cajal-Retzius neurons, nor neurons of the inferior olive and dentate nuclei (Sarnat, Nochlin & Born, Brain Dev. 1998;20(2):88-94. http://www.ncbi.nlm.nih.gov/pubmed/9545178). It's a very useful marker but like any tool, you need to know how to use it.

NeuN is a specific marker for neurons. Indeed NeuN refers to Neuronal nuclear protein.

Hi Eduardo,

I completely agree with Ruben and Baptiste, in normal conditions NeuN is a marker expressed in neurons and not in other cells like glia. Be aware that the expression of neuronal and glial markers can be dysregulated under pathological conditions.

Best,

Ale

If you're using NeuN as a marker you should be aware of its nature and limitations.

Firstly, it's actually the DNA splicing regulator Fox-3 (Kim, Adelstein & Kawamoto, J Biol Chem. 2009;284(45):31052-61. doi: 10.1074/jbc.M109.052969) and NeuN antibodies cross-react with the synaptic protein synapsin I.

Secondly, NeuN is specific to neurons, but not all neurons express NeuN; specifically, it is not expressed by cerebellar Purkinje neurons, olfactory bulb mitral cells (Mullen, Buck & Smith, Development. 1992;116(1):201-11. http://www.ncbi.nlm.nih.gov/pubmed/1483388), sympathetic chain ganglion neurons, cortical Cajal-Retzius neurons, nor neurons of the inferior olive and dentate nuclei (Sarnat, Nochlin & Born, Brain Dev. 1998;20(2):88-94. http://www.ncbi.nlm.nih.gov/pubmed/9545178). It's a very useful marker but like any tool, you need to know how to use it.

Although this is gene expression and not protein levels, I have some preliminary results about expression of NeuN in different types of brain cells (attached). Based on mouse data, NeuN expression is not neuron specific ( it is also expressed in oligodendrocytes) and definitely is not expressed in many neuron types (e.g. dopaminergic neurons, golgi, basket cells, some GABAergic neurons and as previously mentioned  purkinje cells.

Here's a way to see gene expression across many cell types in brain. The data here used is the same as L. Toker's, and compiled from different datasets. It is limited to genes that are on Affymetrix Mouse Expression 430A Array (GPL339)

neuroexpresso.org

neuroexpresso.org

A combination of at least 2 to 3 different staining of neuronal markers need to be always used to confirm the immunohistochemical analysis of neurons in addition to Cresyl violet (Nissel Body) staining. One can use any combination from the list below: 

 b-tubulin III , microtubule-associated protein-2 (MAP-2), neuronal nuclei (NeuN),

neurofilaments (NF) , NF [PAN, clone: DA2/FNP7/RmdO20.11 (PAN)] , and neuron-specific enolase (NSE).

Hi Dr.Lillalch Toker,

I find your plot really interesting. Did you publish it? Can you be more precise about how you performed the experiments?  How did you measure Rbfox3 and Eno2 expression? What are the values of the axis?

Thanks a lot !

Sergio

Hi Sergio,

Thanks for your interest. The data is coming from a curated and normalized collection of cell type specific expression datasets (coming from microarray experiments from different labs). The Y axis represents log2 expression value (just to get an idea, the maximal signal you can get is 14, an in general, 6 is roughly the noise level). You can access all the data through neuroexpresso.org. We just submitted the manuscript, but there is already a version of it on bioRxive: http://biorxiv.org/content/early/2016/11/26/089219

Let me know if you have questions! :)

Lilah

Oh sweet!! Thanks a lot !!

S.

For specification to neuronal cell NeuN is really good one. Researchers have now identified NeuN as Fox-3, a protein involved in regulation of mRNA splicing. Because Fox-3 is expressed exclusively by neurons, suggesting role in regulating neural cell differentiation and nervous system development rather than other cells like astrocytes, microglia

 this is the link  for the paper:

Identification of Neuronal Nuclei (NeuN) as Fox-3, a New Member of the Fox-1 Gene Family of Splicing Factors*

Kee K. Kim, Robert S. Adelstein, and Sachiyo Kawamoto

NeuN was not able to stain pseudorabies virus infected neurons.

Does anyone know if cells in the arcuate nucleus of the hypothalamus are supposed to express NeuN? I see robust labelling in the ventromedial hypothalamus, but very little expression in the arcuate. I would like to know if this is the expected expression pattern. Thank you!