Activity Blockade and GABAA Receptor Blockade Produce Synaptic Scaling through Chloride Accumulation in Embryonic Spinal Motoneurons and Interneurons
Figure 2
Clomeleon labeling in the chick spinal cord provides stable FRET ratios over time.
A) FRET ratios for motoneurons <1 hour after dissection, and >12 hours after dissection at room temperature. For 7 cords (70 cells) there was no statistical difference between the means at each time period (p = 0.1) indicating stability of the signal. Red lines indicate average values. B) Ratios measured over 180 minutes in 27°C Tyrode's solution indicating the stability of the average ratios of 10 cells from 1 cord (black) and for ratios of 5 individual cells (matching colors for B, C, and D). Inset: Average ratios over 180 minutes for 40 cells from 4 cords. C) Subtracted CFP intensity from same 5 cells (avg in black) as in B. D) Subtracted YFP intensity from same 5 cells (avg in black) in B. The stability of the ratios and subtracted CFP and YFP values suggest our imaging methods produce minimal photobleaching.