I am thinking on using NeuN? Any comments, suggestions?
Hello, I think both Tuj1 and NeuN could be good. Many papers us them. On the other hand, what is the system you are going to analyze? Tissue? Cells? from where? Some tissues have more specific markers that label popolations of cells rigth after they are born from progenitors. I can say that in the neocortex Tbr1 is a good marker as it labels the first born neurons.
Immature, newborn neurons/neuroblasts can be stained with DCX, although I suggest Neun for ease of counting. Neun stains embryonic tissue well . It stains the nucleus. There is a similar question on this board that has many fine answers.
Hi thanks for your answers. I am aiming to study newly born neurons and their phenotype specification in the hypothalamic area in tissue slices.
Will you use Brdu,Ki67 or another marker to stain newborn cells?if so, double labeling with DCX or PSA-NCAM will reveal newborn neuroblasts.Labeling with NeuN will stain immature and mature neurons as well whereas calbindin is only expressed from fully matured neurons.If you do not intend to use Brdu or Ki67, there are papers showing that DCX alone can be used as a marker of newborn neurons.
I am intending to use Ki67/PCNA in combination with a neuronal marker (DCX,Hu or NeuN). Could I appply the same technique for animals at different developmental stages? I am working with animals from birthday to adulthood.
Lots of very good suggestions above. Tuj1 (which labels B3 tubulin as I recall), DCX, and NeuN are all widely expressed by neurons and get turned on early after fate commitment so they can be extremely useful markers for cells committing to a neuronal fate. However, it is important to remember that Tuj1, DCX and NeuN may not be expressed by all types of neurons in a given region of the CNS. As an example, there are populations of cells within the retina that do not express B3 tubulin, DCX, or NeuN, which would lead to an undercounting of neurons if any one of those markers was used alone. NeuN also may not be expressed at uniform levels within a brain region, which can make imaging and segmenting an image to quantify cells a bit more of a challenge as well. For example, in our hands at least, cells in the dentate gyrus of the hippocampus show much more intense NeuN labeling that cells in CA1, CA2, or CA3. We've observed this in several other areas of the brain, too.
It might be worthwhile to dig into the available literature on the hypothalamus to see if anyone has identified a ubiquitous neuronal marker specifically for the area you are interested in. Another thing to consider might be to perform double or triple labeling with multiple, widely-expressed neuronal markers to maximize the chance that you won't miss any of these cells in your analyses.
Neuron-specific Class III β-tubulin (TuJ1) is present in newly generated immature postmitotic neurons..so TuJ1 would be best marker for newborn neurons where as NeuN and MAP-2 is for different developmental stages.
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