Pyramidal Neurons - Staining ? (Aug/30/2004 )

Hi there,

I was wondering if anyone knows of a method of staining pyramidal neurons specifically. I need to confirm that I am looking at pyrimidal neurons in the Hippocampus.

I would prefer to do histology to do this however if anyone knows of an antibody that woud be good aswell.

this work will be done on mouse tissue !


Thanks

Find a antibody against a neuronal marker like NeuN, some NMDA receptor subtype, or syaptic protein. Or inject the cell with lucifer yellow or neurobiotin (followed by fluorescent labeled streptavidin) to reveal the neuronal morphology.

Doug,

I found the forum today so i hope you have solved your problem since August. nevertheless, on frozen slices of rat brain, I had very good results with the anti-glutamate transporter EAAC1; The rabbit antibody is available through Alpha diagnostic (San Antonio TX). It works wonderfully at least up to 1:1000. It allows you to identify cells able to transport glutamate. Morphology and location should help you to identify pyramidal cells, since gabaergic interneurons will also label (to a lesser extent though), gaba being synthesized from glutamate . This you can discriminate by doing in parallel a slide with the anti-GAD-67 (careful! the cytoplasmic isoform!) from chemicon. you can also try co-labeling, the anti-GAD is a monoclonal.
Both antibodies will work with PAF fixation+ triton perm.
hope this help

gi