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These results suggest that a smaller fraction of entering calcium ions is captured by the endogenous buffer than in dendrites, which will most likely have a strong impact on the free calcium concentration reached during an AP.
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Next, we tested the pH-hypothesis independent of the use of artificial buffers by modifying the endogenous buffer system.
We used the pH-nose of the Ca2+-current in bipolar cells and cones to determine whether the inhibition of the endogenous buffer system by application of benzolamide was effective.
Added buffer capacity κD, through loading with the calcium dye, was calculated for the various OGB-1 concentrations using equation 3. The inverse of Δ[Ca2+]AP was plotted against κD in figure 2D in order determine the endogenous buffer capacity κE by back extrapolation to the horizontal axis crossing using the linear relation between (Δ[Ca2+]AP)−1 and κD in equation 4 [12] [12].
The buffer capacity experiments indicated that the endogenous buffer capacity in spines is about 3-fold lower than in dendrites.
First we varied the dissociation constant KD (0.5 20 µM) and the total buffer concentration Btot (25 1000 µM) for the endogenous buffer (Figure 5A C).
The observed buffer capacity of 19 for neocortical spines was very similar to the endogenous buffer capacity of ∼20 found in hippocampal spines and small dendrites [8].
The endogenous buffer concentration in dendrites was 660 µM, whereas in spines it was 210 µM (Table 1).
"... Using computer simulations, we study the magnitude of synaptic facilitation (SF) that can be achieved by a buffer saturation mechanism (BSM), and explore its dependence on the endogenous buffering properties.
We assumed that 10 µM of the total endogenous buffer concentration was calmodulin [26] both in spines and dendrites.
The flux rates were much higher than the rates of [Ca2+]i change shown in Fig. 2E because the model includes the effects of strong Ca2+ binding in the cytoplasm by endogenous buffers and fluorescent indicators.
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