I am trying to characterise GABA developmental switch in iPSC derived neurons using Ca2+ imaging. However, as I add GABA agonist, I observe the spontaneous activity of several neurons being abolished, and addition of gabazine in those neurons brings up the transients again.
I am only able to find fewer neurons in culture that show Ca2+ elevation upon GABA addition.
Any idea why this could be happening?
It appears that GABA in inhibitory in the majority of your cultures and you are inhibiting network activity. Adding a GABA agonist likely inhibits excitatory neuron activity which leads to the decrease in spontaneous activity (which is driven by excitatory network activity), and the addition of gabazine removes this inhibition and allows the excitatory network activity to return. Have you tried glutamate receptor antagonists and seen the same response?
Aaron G Roseberry Hi, thanks for replying!
No, I haven't yet tried glutamate receptor antagonists. I do plan to carry out these experiments again with AP-5+CNQX and look at the calcium profile.
I found one explanation from Anthony van de Paul 1995, reporting similar findings in mouse hypothalamic cultures.
One of the possible explanation is that chloride transporters (NKCC1, KCC2) are differentially expressed within the same culture.
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