I have various data on calcium dynamics. Generated graph are look oscillatory. I really want to know what are things i can predict from these graph and analyses it completely. For support i am attaching one graph.
Hi
Might you give me a little bit more info about your protocol. Beside it would be interesting to know what cells are you recording, are they in vitro cultures?, how many days in vitro?...
have a great day
Felipe
quite interesting... I do teh same cultures. So there you have two records. In the top one, I don´t see any spontaneous activity (Calcium spikes), on the other hand, the spontaneous spikes are quite obvious. This depends on the variability of the cells in culture, as far as I understand neurons wich are a great mistery. I´m not quite sure what baclofen does, but whatever you can see an intantaneous increment in the spikes of Calcium, which result to me quite normal.
Something else you would see in the one below, is how the spontaneous spiking decreses over the time... I´m not quite sure why this happen, but I guess it might be due the protocol.. are these experimets done with a perfusion system? what kind of buffer are you using?
Are you recording from a single neuron or from an area of the coverslip? Also, what is the difference between the top and the bottom trace? A bit more info will help us help you.
Both are neuron cell in single culture plate where at 100 time frame baclofen drug was added of 20um.
Dear Sarpras,
GABAb receptoras are predominatly inhibitory and some directly couple to VSCCs via Gi/Go. You would expect baclophen to inhibit spontaneous spiking or other receptors (muscarinic, alpha-adrenergic, phosphatidic acid, etc) induced calcium transients and/or fluctuations. Your first trace seems to indicate that baclophen does not induce Ca fluctuations (as one would expect). The second trace shows spontaneous spiking and the addition of baclophen might have partially inhibited it, but you need: 1- a much longer pre-baclophen recording to assess basal spontaneous activity; 2-statitstics from a large number of cells to prove your point. You might also look at baclophen effects on potassium-induced depolarization and Ca entry/spiking.
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