TRPP2 and STIM1 form a microdomain to regulate store-operated Ca2+ entry and blood vessel tone
| Autor: | Jie Li, Jizheng Guo, Bing Shen, Muyao Zhou, Jiexia Chen, Xiaoqiang Yao, Ren Zhao, Juan Du |
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| Rok vydání: | 2019 |
| Předmět: |
inorganic chemicals
Small interfering RNA Vascular smooth muscle Thapsigargin lcsh:Medicine Biochemistry 03 medical and health sciences chemistry.chemical_compound Vascular smooth muscle cells lcsh:QH573-671 education Polycystin-2 Molecular Biology 0303 health sciences education.field_of_study lcsh:Cytology lcsh:R 030302 biochemistry & molecular biology Stromal interaction molecule 1 STIM1 Cell Biology Transfection Inositol trisphosphate receptor Cell biology Polycystin 2 chemistry Store-operated Ca2+ entry Blood vessel tone Adenosine triphosphate |
| Zdroj: | Cell Communication and Signaling, Vol 18, Iss 1, Pp 1-16 (2020) |
| DOI: | 10.21203/rs.2.18317/v1 |
| Popis: | Background Polycystin-2 (TRPP2) is a Ca2+ permeable nonselective cationic channel essential for maintaining physiological function in live cells. Stromal interaction molecule 1 (STIM1) is an important Ca2+ sensor in store-operated Ca2+ entry (SOCE). Both TRPP2 and STIM1 are expressed in endoplasmic reticular membrane and participate in Ca2+ signaling, suggesting a physical interaction and functional synergism. Methods We performed co-localization, co-immunoprecipitation, and fluorescence resonance energy transfer assay to identify the interactions of TRPP2 and STIM1 in transfected HEK293 cells and native vascular smooth muscle cells (VSMCs). The function of the TRPP2-STIM1 complex in thapsigargin (TG) or adenosine triphosphate (ATP)-induced SOCE was explored using specific small interfering RNA (siRNA). Further, we created TRPP2 conditional knockout (CKO) mouse to investigate the functional role of TRPP2 in agonist-induced vessel contraction. Results TRPP2 and STIM1 form a complex in transfected HEK293 cells and native VSMCs. Genetic manipulations with TRPP2 siRNA, dominant negative TRPP2 or STIM1 siRNA significantly suppressed ATP and TG-induced intracellular Ca2+ release and SOCE in HEK293 cells. Inositol triphosphate receptor inhibitor 2-aminoethyl diphenylborinate (2APB) abolished ATP-induced Ca2+ release and SOCE in HEK293 cells. In addition, TRPP2 and STIM1 knockdown significantly inhibited ATP- and TG-induced STIM1 puncta formation and SOCE in VSMCs. Importantly, knockdown of TRPP2 and STIM1 or conditional knockout TRPP2 markedly suppressed agonist-induced mouse aorta contraction. Conclusions Our data indicate that TRPP2 and STIM1 are physically associated and form a functional complex to regulate agonist-induced intracellular Ca2+ mobilization, SOCE and blood vessel tone. |
| Databáze: | OpenAIRE |
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