• Authors: Ramesh G. et al.
  • Year: 2021
  • Journal: Cell Rep 34(11)
  • Applications: in vitro / DNA / jetOPTIMUS, jetPRIME
  • Cell types:
    1. Name: HEK-293
      Description: Human embryonic kidney Fibroblast
      Known as: HEK293, 293
    2. Name: MEF
      Description: Murine embryonic fibroblast cells 
    3. Name: SH-SY5Y
      Description: Human neuroblastoma cells
      Known as:


For Bimolecular fluorescence complementation, HEK cells were transfected using jetOPTIMUS: 1.2x 10^5 HEK STIM1−/−;STIM2−/− cells were seeded in 6 well plates and transfected with a total amount of 2 μg DNA. For Fura2-based Ca2+ Imaging, jetPRIME was used: - MEF cells were seeded on 25mm coverslips 24 hours before transfection with 2μg of plasmids - SH-SY5Y cells were seeded in 35mm cell culture dishes 24 hours before transfection with 2μg of plasmids. Transfected SH-SY5Y were reseeded 4 hours post transfection on 25mm coverslips.


Store-operated Ca2+-entry (SOCE) regulates basal and receptor-triggered Ca2+ signaling with STIM proteins sensing the endoplasmic reticulum (ER) Ca2+ content and triggering Ca2+ entry by gating Orai channels. Although crucial for immune cells, STIM1's role in neuronal Ca2+ homeostasis is controversial. Here, we characterize a splice variant, STIM1B, which shows exclusive neuronal expression and protein content surpassing conventional STIM1 in cerebellum and of significant abundance in other brain regions. STIM1B expression results in a truncated protein with slower kinetics of ER-plasma membrane (PM) cluster formation and ICRAC, as well as reduced inactivation. In primary wild-type neurons, STIM1B is targeted by its spliced-in domain B to presynaptic sites where it converts classic synaptic depression into Ca2+- and Orai-dependent short-term synaptic enhancement (STE) at high-frequency stimulation (HFS). In conjunction with altered STIM1 splicing in human Alzheimer disease, our findings highlight STIM1 splicing as an important regulator of neuronal calcium homeostasis and of synaptic plasticity.