We have robust staining for a certain ion channel that is located in the neuropil and wish to know whether its axonal or dendritic. Hoping to use double immunostaining to find out.
Hi Susan,
a reviewer has asked why i had not used MAP2 to confirm the neurites i was looking at were dendrites. Below is the whole answer i gave them, with in bold some interesting references.
hope that helps.
"In the early phase of neuronal differentiation, one neurite sprout is selected to become the axon based on its spatial relationship to the Golgi and centrosome. Subsequently, the remaining neurites take up a dendritic identity. As a result, the axon is most often, but not always, the longest neurite. To unambiguously identify the axon, we assess several other well characterized axon-specific parameters described by Craig and Banker (1994). These are routinely used and are well accepted within the field (Lanoue et al, 2013). We identify the axon according to the following parameters:
i. the longest neurite
ii. the caliber of the axon adjacent to the cell body is smaller than that of the dendrite,
iii. the diameter of the dendrite decreases at each branch point, whereas the axonal diameter remains constant,
iv. the angle between the primary neurite and the branch is smaller in dendrites compared to that in axons.
Therefore, it is not necessary to stain for MAP2 (for a review of this topic see Cáceres et al, 2012). Throughout this study we analyzed only those neurons for which the axon could be unambiguously identified.
Note: MAP2 is present in the axon initial segment, but is not found in the axon past this point (Yap et al, 2016, Wang et al, 2016)."
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