calcium increases - (Jul/01/2006 )

I do not know if somone is familiar with calcium responses. I am using a CHO-K1 cell line transfected with several independent GPCRs to study calcium increases after agonist stimulation.
I observe what I consider "normal" responses at only 14 sec form agonist stimulation, but using a different GPCR, I see other kind of stimulatory responses, this time very delayed at about 40 of 50 sec from stimulation. Do someone kows about which could be the mechanism implied in that late response? I would be very pleased.
Thanks in advance.

Hi,
Sorry for not having a response for u but rather a counterquestion
I am also trying to study calcium levels in fibroblasts
what method do u use? is it confocal?
what is gpcr and what agonist u are using?

could it be that u are not using the right agonist for ur gpcr?

Hi,
Sorry for not having a response for u but rather a counterquestion
I am also trying to study calcium levels in fibroblasts
what method do u use? is it confocal?
what is gpcr and what agonist u are using?

could it be that u are not using the right agonist for ur gpcr?



Hello!
I am working with somatostatin receptors and using SRIF-14 and CST-17 as agonists.
We employ a widefield Nikon microscope and Fura-2 as probe and it usually works very good.
What are you going to use?

I was using calcium green and tried both FACS and confocal to try measure steady state levels of calcium. I was advised to measure calcium responses instead steady states levels and I discarded FACS analysis for this as i believe its not appropriate. My cells are transduced fibroblasts with calcium related hormones (we still dont know for sure)

tertu