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. 2021 Jan 5;12:573714. doi: 10.3389/fnsyn.2020.573714

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Copyright © 2021 Kushnireva, Korkotian and Segal.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Motility of STIM1 puncta along a dendrite and their association with a transient rise of [Ca²⁺]_i. (A) Three images of a dendritic segment containing one stable STIM1 puncta, and a mobile one moving from left to right. (B) Fluo2 sensor shows calcium transients. Panels (B1–3) show calcium transient in the area, marked with an arrowhead. The current location of the mobile STIM1 punctum is marked with a red contour. The second red contour corresponds to the stable STIM1 punctum. (C) When punctum is present at the red location on the right a calcium transient can be detected. (D) In an adjacent location, neither puncta nor calcium transient is seen. Averaged data, n = 10 regions from different cells and different cultures. Three time points are taken: right before the entrance of puncta into the selected region (before) in the presence of puncta within the region of interest (during) and right after the puncta left the region (after). Simultaneously, measurements from a randomly selected nearby region were made (nearby). STIM1 for these time points (E, red). Nearby region (E, gray). Calcium for these time points (F, dark green). Calcium measurements from a nearby region (F, light green). (G) Time course of change in [Ca²⁺]_i fluorescence at the STIM1 puncta (top, green) and the STIM1 fluorescence (red, low trace) of 10 dendrites. Note that STIM1 fluorescence precedes the rise of [Ca²⁺]_i and is sustained after the calcium change subsided. (H) A fast synapse-evoked [Ca²⁺]_i rise that is not associated with a change in STIM fluorescence. Note the much faster time course of change in the synaptic event compared to the change seen above. (I) Movement rates (μm/50 s of three sizes of STIM1 puncta (large: >2 μm, medium: 1–2 μm and small: <1 μm), at three age groups, 10, 15 and 20 DIV. Small puncta move faster than medium and large ones at all age groups (p < 0.05). The faster movement rate is observed at 15 DIV compared to 10 and 20 DIVs for both medium and small-sized STIM1 puncta (medium: n = 27, 19 and 17 for 10, 15 and 20 DIV, respectively, F = 3.58, p < 0.05; small: n = 27, 14 and 8, F = 3.02, p < 0.05). *Significant, 0.05 > p > 0.01; **very significant, 0.01 > p > 0.001; n.s., not significant, p ≥ 0.05.