I am differentiating Neural stem cells into neurons. I have planned to use Tau and Map2 antibodies in western blotting to differentiate axon and dendrite outgrowth. I would like to know any other experiments which could be used to differentiate the axon or dendrite outgrowth of a neuron other than western blot and ICC.
Hi Karthikeyan,
For ICC, another classic marker is AnkyrinG: it will label the AIS (Axon Initial Segment) and thus clearly identifies the axon.
In electrophysiology we mainly use visual cues to identify the axon: a thin, aspiny process, with a regular diameter. It is often really tortuous and has small varicosities along the way, which are presynaptic boutons.
In comparison, dendrites are much bigger when starting from the soma, they fork regularly and they have spines (in the case of excitatory neurons).
The best way would be to differentiate them functionally, so you prove they do play the role of an axon or dendrite. But to do so you will have to record the cell by patching the soma and either do extracellular recording with another pipette, or do some functional imaging in the boutons and/or dendrites.
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