title: Effects of stromal interaction molecule 1 or Orai1 overexpression on the associated proteins and permeability of podocytes.
abstract: OBJECTIVE
  The present study was conducted to determine whether two important signalling molecules of store-operated channel (SOC), stromal interaction molecule 1 (STIM1) and Orai1, were involved in glomerular podocyte injury. We explored the effects of STIM1/Orai1 overexpression on podocyte associated proteins and cell permeability.
  
  
  METHODS
  The expression of STIM1 and Orai1 were examined in the renal cortex of adriamycin-induced nephropathy mice by real-time RT-PCR. The recombinant plasmid of STIM1/Orai1, identified by restriction enzyme digestion and PCR, was transfected into MPC5 cells via lipofectamine 2000. The transfecting efficiency was observed by a fluorescence microscope. RT-PCR and Western blotting were used to evaluate the expression levels of STIM1, Orai 1 and some podocyte-associated molecules in the transfected MPC5 cells. In addition, we examined the diffusion of FITC-dextran across the podocyte monolayer to investigate whether STIM1/Orai1 overexpression could affect cell permeability.
  
  
  RESULTS
  We found that the mRNA levels of STIM1 and Orai1 were increased in adriamycin-induced nephropathy mice. STIM1/Orai1 overexpression significantly decreased the expression of podocin and CD2-associated protein (CD2AP), whereas it increased the expression of α-actinin-4. The permeability was significantly increased in the STIM1/Orai1 overexpression group.
  
  
  CONCLUSIONS
  Our findings suggested that STIM1/Orai1 overexpression could affect the cell permeability and the expression of partial podocyte-associated proteins, which may ultimately result in podocyte injury.
authors.affiliation: Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.|Lianyungang Clinical School of Nanjing Medical University, Lianyungang, China.
authors.forename: Li
authors.initials: L
authors.lastname: Miao
authors.affiliation: Lianyungang Clinical School of Nanjing Medical University, Lianyungang, China.
authors.forename: Dongyue
authors.initials: D
authors.lastname: Wei
authors.affiliation: Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Lianyungang Clinical School of Nanjing Medical University, Lianyungang, China.
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Liu
authors.affiliation: Lianyungang Clinical School of Nanjing Medical University, Lianyungang, China.
authors.forename: Siguang
authors.initials: S
authors.lastname: Lu
authors.affiliation: Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.
authors.forename: Aihua
authors.initials: A
authors.lastname: Zhang
authors.affiliation: Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China. smhuang1966@163.com.
authors.forename: Songming
authors.initials: S
authors.lastname: Huang
mesh_terms: D000818:Animals; D002463:Cell Membrane Permeability / Q000502:physiology; D020869:Gene Expression Profiling / Q000379:methods; D007672:Kidney Cortex / Q000378:metabolism; D007674:Kidney Diseases / Q000378:metabolism / Q000473:pathology; D051379:Mice; D023421:Models, Animal; D000071740:ORAI1 Protein / Q000378:metabolism*; D050199:Podocytes / Q000378:metabolism*; D000071737:Stromal Interaction Molecule 1 / Q000378:metabolism*; D014162:Transfection / Q000379:methods
chemical_list: D000071740:ORAI1 Protein; D000071737:Stromal Interaction Molecule 1
keywords: Orai1; STIM1; cell permeability; podocyte-associated proteins