Fig. 1. Identification of the components of SOCE machinery in neurons and SGC units of rat sympathetic ganglia.
(A, a) H&E staining to demonstrate the close localization of SGCs around the large soma of an SCG neuron. (A, b) Kir4.1-positive SGCs (arrows) enveloping SCG neurons (n). (A, c-d) DIC and immunofluorescent images of a neuron-SGC unit in the culture. SGCs showed Kir4.1-IR (arrow). DAPI was used to stain the nuclei of neuron and SGC. Scale bars = 10 μM. (B) Relative mRNA expressions of Orai and STIM, the SOCE components in sympathetic ganglia. The number of experiments is indicated in parentheses. (C) Representative immunofluorescent images of Orai1 (green) and STIM1 (red) distribution in a sympathetic neuron-SGC unit. DAPI was used to stain the nuclei of both neuron and SGC. Confocal images of the cells were captured before and after a 10-min treatment with 30 μM CPA. ER Ca2+ depletion with CPA induced the mobilization of STIM1-Orai1 aggregates, resulting in the assembly of prominent puncta formation in the cell surfaces of a neuron-SGC unit. Scale bars = 10 μM. (D, E). Summary of the number of Orai1 puncta in neuron-SGC units. Vehicle DMSO-treated (VEH) and CPA-treated (CPA) groups were compared (n = 54–60 cells from 5 wells per group). The data are presented as means ± SEM. Unpaired t-test, ***p < 0.001. SOCE, store-operated calcium entry; SGC, satellite glial cell; SCG, superior cervical ganglia; DIC, differential interference contrast; DAPI, 4,6-diamino-2-phenylindole dihydrochloride; STIM, stromal interaction molecule; CPA, cyclopiazonic acid; ER, endoplasmic reticulum; DMSO, dimethyl sulfoxide.
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