Cellular and Molecular Life Sciences

, Volume 70, Issue 17, pp 3211–3227 | Cite as

The unconventional secretion of stress-inducible protein 1 by a heterogeneous population of extracellular vesicles

  • Glaucia N. M. Hajj
  • Camila P. Arantes
  • Marcos Vinicios Salles Dias
  • Martín Roffé
  • Bruno Costa-Silva
  • Marilene H. Lopes
  • Isabel Porto-Carreiro
  • Tatiana Rabachini
  • Flávia R. Lima
  • Flávio H. Beraldo
  • Marco M. A. Prado
  • Rafael Linden
  • Vilma R. Martins
Research Article


The co-chaperone stress-inducible protein 1 (STI1) is released by astrocytes, and has important neurotrophic properties upon binding to prion protein (PrPC). However, STI1 lacks a signal peptide and pharmacological approaches pointed that it does not follow a classical secretion mechanism. Ultracentrifugation, size exclusion chromatography, electron microscopy, vesicle labeling, and particle tracking analysis were used to identify three major types of extracellular vesicles (EVs) released from astrocytes with sizes ranging from 20–50, 100–200, and 300–400 nm. These EVs carry STI1 and present many exosomal markers, even though only a subpopulation had the typical exosomal morphology. The only protein, from those evaluated here, present exclusively in vesicles that have exosomal morphology was PrPC. STI1 partially co-localized with Rab5 and Rab7 in endosomal compartments, and a dominant-negative for vacuolar protein sorting 4A (VPS4A), required for formation of multivesicular bodies (MVBs), impaired EV and STI1 release. Flow cytometry and PK digestion demonstrated that STI1 localized to the outer leaflet of EVs, and its association with EVs greatly increased STI1 activity upon PrPC-dependent neuronal signaling. These results indicate that astrocytes secrete a diverse population of EVs derived from MVBs that contain STI1 and suggest that the interaction between EVs and neuronal surface components enhances STI1–PrPC signaling.


Exosomes Chaperones Prion protein STI1 Extracellular vesicles 



We are very grateful to Dr. Wes Sundquist for the donation of VPS4A plasmids. We thank Leica Microsystems and Ms. Lianne Dale for help in experiments done in a demo Leica GSD unit. This investigation was supported by grants from FAPESP to VRM (2009/14027-2) and GNMH (2012/04370-4), PrioNet-Canada, Canadian Institutes of Health Research (CIHR), Canadian Foundation for Innovation and Ontario Research Fund to MAMP, CNPq and FAPERJ to RL, FAPESP fellowships to CA, MVSD, MR and BS, and a postdoctoral CNPq fellowship to IPC are gratefully acknowledged.

Conflict of interest

The authors declare that there are no conflicts of interest.

Supplementary material

18_2013_1328_MOESM1_ESM.tif (121 kb)
Supplementary Fig. 1 CM collected either from serum free (0 %) astrocytes or from astrocytes cultured in 10 % serum were applied to gel filtration chromatography columns and fractions 1, 2, 5, 6, 11, and 12 were subjected to immunoblotting for STI1 (TIFF 121 kb)
18_2013_1328_MOESM2_ESM.tif (2.5 mb)
Supplementary Fig. 2 a Astrocyte cultures were transfected with GFP and CM was ultracentrifuged for 2 h. Pellets and supernatant were added to primary hippocampal neurons SN-56 cells. Scale bar 10 μm (TIFF 2555 kb) (8.7 mb)
Supplementary movie 1 STI1-GFP was co-transfected in astrocytes with mCherry Rab7 and live imaged in a TIRF microscope. Total time 3 min (MOV 8907 kb)
18_2013_1328_MOESM4_ESM.avi (4 mb)
Supplementary movie 2 STI1-GFP was co-transfected in astrocytes with mCherry Rab5 and live imaged in a TIRF microscope. Total time 5 min (AVI 4065 kb)


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Copyright information

© Springer Basel 2013

Authors and Affiliations

  • Glaucia N. M. Hajj
    • 1
    • 2
  • Camila P. Arantes
    • 3
  • Marcos Vinicios Salles Dias
    • 1
    • 2
  • Martín Roffé
    • 1
    • 2
  • Bruno Costa-Silva
    • 1
    • 2
  • Marilene H. Lopes
    • 4
  • Isabel Porto-Carreiro
    • 5
  • Tatiana Rabachini
    • 6
  • Flávia R. Lima
    • 7
  • Flávio H. Beraldo
    • 8
  • Marco M. A. Prado
    • 8
  • Rafael Linden
    • 5
  • Vilma R. Martins
    • 1
    • 2
  1. 1.International Research CenterA.C. Camargo HospitalSão PauloBrazil
  2. 2.National Institute for Translational Neuroscience and National Institute of OncogenomicsSão PauloBrazil
  3. 3.Department of Biochemistry, Chemistry InstituteUniversity of São PauloSão PauloBrazil
  4. 4.Department of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  5. 5.Instituto de Biofisica Carlos Chagas FilhoFederal University of Rio de JaneiroRio de JaneiroBrazil
  6. 6.Ludwig Institute for Cancer ResearchSão PauloBrazil
  7. 7.Instituto de Ciências BiomédicasFederal University of Rio de JaneiroRio de JaneiroBrazil
  8. 8.Department of Anatomy and Cell Biology and Department of Physiology and Pharmacology, Robarts Research InstituteUniversity of Western OntarioLondonCanada

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