Currently I am working in the area of adult neurogenesis. I am using Golgi-Cox and Scholl's method to analyse dendritic arborization (though its not specific for newborn neurons). Can I use SMI-32 immunohistochemical method to analyze dendritic arborization of the neurons (neurons- including newborn and already exisiting too)? It would be helpful for me if I get any other suggestions.
Hi Rajesh,
I work on adult hippocampal neurogenesis so I can help you a little bit. The best way to analyze dendritic arborization of newborn neurons is to use a retroviral vector that express a fluorescent protein (e.g. GFP) and inject it in the dentate gyrus of anesthetize mice. Use a stereotaxic frame to be precise when injecting your retrovirus. Retroviral vectors will integrate in dividing stem/progenitor cells in the adult dentate gyrus and will be useful as a lineage tracing. You need to decide how long you want to keep your mice before sacrifice them to analyze a particular developmental stage (e.g. 28 days post injection). You can analyze the morphology of the dendritic arborization of newborn neurons easily by staining your brain slices for the fluorescent protein. Once you have the stained brain slices, remember to use an appropiate software (NeuronJ, plugin for ImageJ) to analyze your z-stack files. You can measure total length of dendritic arborization, number of ramifications, etc.
Hope that helped to your research, and remember to have all your AUFs approved from your IACUC committee to perform these experiments ahead of time.
Best,
Ale
Try Lucifer yellow or brown in liposomes and inject: see Thunnissen and Marani, J neurosci Methods. Don't have the article at hand, look in my publication list of research gate.
Check out this article!
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0033114
Describes a method called ImmunoGolgi which combines the Golgi-Cox method with immunofluorescence labeling of new neurons (with BrdU).
Then, you might be able to specifically determine the dendritic arborization of new born neurons.
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