Changes in NKCC1 expression are seen in in vivo models of epilepsy and brain tissue from patients with drug-resistant seizures. Is it possible to study such changes using acute hippocampal slice preparations (using western blotting)?
More precisely, if I incubate hippocampal slices in zero Mg2+ or high K+ conditions for 1-3 hours, will that trigger changes in NKCC1 expression already?
I am interested to know how long it would take to regulate NKCC1 expression.
If you are trying to find conformation change use immuno precipitation and mass spec and phosphorylation specific antibodies in immunoblot
From the gene expression perspective, it is possible. But there is no guarantee to demonstrate such a change in acute model. Generally, I think that you need a chronic model like Pilocarpine to see that.
Yes, that's possible. May depend on what and how old your in vivo models were, and what drugs, for how long.. were tried among the drug-resistant patients. Will be nice to hear your results.
Please see my publications in pubmed "Jayakumar AR" (NKCC1 expression, phosphorylated, oxidized and nitrated forms by western blots, JBC 2008; J Hepatol, 2011, J Neurochem 2011 and others).
Dear Arumugam,
Thank you for the interesting references. From your articles, I realize you have studied NKCC1 expression in cultured cells. Do you think it would be different for acute slice preparations? Thank you.
Please read carefully my J Hepatol paper 2011; in MM "Immunoblots of NKCC1" and Figs 2, 3 and 4 (3 and 4 are NKCC1 protein levels in rat brain cortiacl tissues and 2 is the NKCC activity). Please send email to [email protected] if you suppose need more informations or the J Hepatol paper.
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