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. 2022 Mar 23;11:e76993. doi: 10.7554/eLife.76993

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© 2022, Landau et al

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Figure 1. — (A) Maximum z-projection of Alexa 594 fluorescence showing the full apical dendritic morphology of a cortical L2/3 pyramidal cell. Two sites corresponding to the dendritic regions with low (blue) and high (black) ΔCa_AP are indicated. (B) Frame scan (left) showing high ΔCa_AP branch. Dotted yellow line indicates the orientation of the line scan used to acquire the data in the kymograph (right) of Fluo-5F fluorescence in the spine and neighboring dendrite evoked by a bAP. Star indicates glutamate uncaging location. White vertical lines indicate ROIs for spine and dendrite. Inset: bAP waveform recorded at the soma. (C) Average calcium-dependent fluorescence transients in a high ΔCa_AP dendritic spine (left) and parent dendrite (right) evoked by 1 bAP, an uEPSP, and pairing of an uEPSP with 1 bAP (bAP evoked 5ms after the uEPSP). Inset: average somatic whole-cell recording of the uEPSPs. In this and all presentations of imaging data, the red shaded area of the spine and dendritic schematic indicates the region from which fluorescence was measured. (D) As in B for the low ΔCa_AP branch. (E) As in C for the low ΔCa_AP branch. Note the difference in ΔCa_AP (black) between the spines in B-C and D-E. bAP-evoked calcium influx and bAP-dependent amplification are decoupled in L2/3 pyramidal cells.

Figure 1—figure supplement 1. — (A) Maximum z-projection of Alexa 594 fluorescence showing the full apical dendritic morphology of a cortical L2/3 pyramidal cell. Two sites corresponding to the dendritic regions with low (blue) and high (black) ΔCa_AP are indicated. (B) Frame scan (left) showing high ΔCa_AP branch. Dotted yellow line indicates the orientation of the line scan used to acquire the data in the kymograph (right) of Fluo-5F fluorescence in the spine and neighboring dendrite evoked by a bAP. Star indicates glutamate uncaging location. White vertical lines indicate ROIs for spine and dendrite. Inset: bAP waveform recorded at the soma. (C) Average calcium-dependent fluorescence transients in a high ΔCa_AP dendritic spine (left) and parent dendrite (right) evoked by 1 bAP, an uEPSP, and pairing of an uEPSP with 1 bAP (bAP evoked 5ms after the uEPSP). Inset: average somatic whole-cell recording of the uEPSPs. In this and all presentations of imaging data, the red shaded area of the spine and dendritic schematic indicates the region from which fluorescence was measured. (D) As in B for the low ΔCa_AP branch. (E) As in C for the low ΔCa_AP branch. Note the difference in ΔCa_AP (black) between the spines in B-C and D-E. bAP-evoked calcium influx and bAP-dependent amplification are decoupled in L2/3 pyramidal cells.