Sunday, July 23, 2006
♂ Science is bloody tough..
After so many failures with the GAP-43 antibody on thalamus sections, I have made a decision. I am not going to follow the protocols that the Research assistant in my lab has used. I am going to try come up with my own dilutions for my next batch of brains.
If it works... that's awesome... IF they don't... I'm gonna be really pissed... I guess I just have to stick with my VGLUT2 synaptic count results. However, IF the VGLUT2 counts are negative, the GAP-43 results would most certainly be negative. I mean... IF there's no synapses being formed, it's most probably because there's no sprouting at all... hence, no GAP-43. I guess there's nothing happening in the ventrolateral thalamus at all.
On another note, I've looked at a couple of cerebellar sections and voila! GAP-43 positive cells on the deep cerebellar nuclei!! I guess I have to be content with a negative result for my thalamus project(honours). However, if I'm going to go on and do PhD, I will be looking at the deep cerebellar nuclei in more detail. I strongly believe that there's some significant sprouting in the deep cerebellar nuclei. The question is... where do they come from??? Purkinje Cells?? or somewhere else??
However, before I answer those questions... I need to know the best way of cutting these cerebellar sections.. because they are so fragile.. The reason that I do not have that many cerebellar data is because most of my cerebellar sections are wrecked. It would be fun also to look at the cerebellum and count the synapses in them. However, I am not sure what kind of synapses are present in the cerebellum. If I'm gonna count the Purkinje synapses, I may have to use a GABA transporter antibody to mark the inhibitory synapses. If I wanna count other synapses, I probably have to do some research on their identity.
11:44 AM
