FURA-2 - (Mar/14/2005 )

Has anybody skills in Fura-2 measurements? We want so mearsure Ca++ in the cytosol of neuronal cells. (We already performed it in endothelial cells). But how to stiumulate Ca++ release from the ER in neuronal cells?

Thank you very much!

Hi there... I've been working with Fura-2-AM for the last 3 years... I think I can help you!!
Ca2+ release is mainly obtained by stimulating the cells with an agonist binding to a GPCR coupled with alpha-q. Muscarinic receptor, CCK, ATP, Ang2, etc... If your cells do not have such receptor, you can induce a calcium leak from the RE using thapsigargin, which blocks the SERCA. The calcium pools the go empty by themselves (not really empty.... just equilibrated with the cytosol)
I suggest you do a quick search on PubMed about this... there's tons of papers treating this subject.

Simon

Hi,

In a previous lab I used Fura-2 to measure Ca++ release in mouse neuronal cultures. We pulsed on glutamate to initiate the release. You should check out Dr. Gregory J. Brewer's work. He is at the Southern Illinois University School of Medicine. I believe they are still working on this study.