Abstract
Stroke is pathologically associated with oxidative stress, protein damage, and neuronal loss. We previously reported that overexpression of a ubiquitin-like protein, ubiquilin-1 (Ubqln), protects neurons against ischemia-caused brain injury, while knockout of the gene exacerbates cerebral ischemia-caused neuronal damage and delays functional recovery. Although these observations indicate that Ubqln is a potential therapeutic target, transgenic manipulation-caused overexpression of Ubqln occurs before the event of ischemic stroke, and it remains unknown whether delayed Ubqln overexpression in post-ischemic brains within a clinically relevant time frame is still beneficial. To address this question, we generated lentiviruses (LVs) either overexpressing or knocking down mouse Ubqln, and treated post-ischemic stroke mice 6 h following the middle cerebral artery occlusion with the LVs before animal behaviors were evaluated at day 1, 3, 5, and 7. Our data indicate that post-ischemic overexpression of Ubqln significantly promoted functional recovery, whereas post-ischemic downregulation of Ubqln expression delays functional recovery. To further understand the mechanisms underlying how Ubqln functions, we also isolated protein aggregates from the brains of wild-type mice or the mice overexpressing Ubqln following ischemia/reperfusion. Western blot analysis indicates that overexpression of Ubqln significantly reduced the accumulation of protein aggregates. These observations not only suggest that Ubqln is a useful candidate for therapeutic intervention for ischemic stroke but also highlight the significance of proteostasis in functional recovery following stroke.
Abbreviations
- MCAO:
-
Middle cerebral artery occlusion
- I/R:
-
Ischemia/reperfusion
- Ub:
-
Ubiquitin
- UPS:
-
Ubiquitin–proteasome system
- Ubqln:
-
Ubiquilin-1
- OE:
-
Overexpression
- KO:
-
Knockout
- shRNA:
-
Small hairpin ribosomal nucleic acid
- LV:
-
Lentivirus
- GFP:
-
Green fluorescence protein
- mNSS:
-
Modified neurologic severity score
- TX:
-
Triton-X100
- SDS:
-
Sodium dodecyl sulfate
- PAGE:
-
Polyacrylamide gel electrophoresis
References
Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD (2009) A versatile viral system for expression and depletion of proteins in mammalian cells. PLoS ONE 4(8):e6529. doi:10.1371/journal.pone.0006529
Chen J, Zhang C, Jiang H, Li Y, Zhang L, Robin A, Katakowski M, Lu M, Chopp M (2005) Atorvastatin induction of VEGF and BDNF promotes brain plasticity after stroke in mice. J Cereb Blood Flow Metab 25(2):281–290. doi:10.1038/sj.jcbfm.9600034
Chen X, Guo C, Kong J (2012) Oxidative stress in neurodegenerative diseases. Neural Regen Res 7(5):376–385. doi:10.3969/j.issn.1673-5374.2012.05.009
Deglon N, Aebischer P (2002) Lentiviruses as vectors for CNS diseases. Curr Top Microbiol Immunol 261:191–209
Dong G, Callegari EA, Gloeckner CJ, Ueffing M, Wang H (2012) Prothymosin-alpha interacts with mutant huntingtin and suppresses its cytotoxicity in cell culture. J Biol Chem 287(2):1279–1289. doi:10.1074/jbc.M111.294280
Ge P, Luo Y, Liu CL, Hu B (2007) Protein aggregation and proteasome dysfunction after brain ischemia. Stroke 38(12):3230–3236. doi:10.1161/STROKEAHA.107.487108
Huang HF, Guo F, Cao YZ, Shi W, Xia Q (2012) Neuroprotection by manganese superoxide dismutase (MnSOD) mimics: antioxidant effect and oxidative stress regulation in acute experimental stroke. CNS Neurosci Ther 18(10):811–818. doi:10.1111/j.1755-5949.2012.00380.x
Liu Y, Hettinger CL, Zhang D, Rezvani K, Wang X, Wang H (2014a) The proteasome function reporter GFPu accumulates in young brains of the APPswe/PS1dE9 Alzheimer’s disease mouse model. Cell Mol Neurobiol 34(3):315–322. doi:10.1007/s10571-013-0022-9
Liu Y, Lu L, Hettinger CL, Dong G, Zhang D, Rezvani K, Wang X, Wang H (2014b) Ubiquilin-1 protects cells from oxidative stress and ischemic stroke caused tissue injury in mice. J Neurosci 34(8):2813–2821. doi:10.1523/JNEUROSCI.3541-13.2014
Lu L, Wang H (2012) Transient focal cerebral ischemia upregulates immunoproteasomal subunits. Cell Mol Neurobiol 32(6):965–970. doi:10.1007/s10571-012-9854-y
Park IH, Arora N, Huo H, Maherali N, Ahfeldt T, Shimamura A, Lensch MW, Cowan C, Hochedlinger K, Daley GQ (2008) Disease-specific induced pluripotent stem cells. Cell 134(5):877–886. doi:10.1016/j.cell.2008.07.041
Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, Sabatini DM, Chen IS, Hahn WC, Sharp PA, Weinberg RA, Novina CD (2003) Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 9(4):493–501
Wang H, Monteiro MJ (2007) Ubiquilin interacts and enhances the degradation of expanded-polyglutamine proteins. Biochem Biophys Res Commun 360(2):423–427. doi:10.1016/j.bbrc.2007.06.097
Wang H, Yu SW, Koh DW, Lew J, Coombs C, Bowers W, Federoff HJ, Poirier GG, Dawson TM, Dawson VL (2004) Apoptosis-inducing factor substitutes for caspase executioners in NMDA-triggered excitotoxic neuronal death. J Neurosci 24(48):10963–10973
Acknowledgements
This work was supported by the National Institute of Neurological Disorders and Stroke under research Grants R03NS084340 and R01NS088084 (HW).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Rights and permissions
About this article
Cite this article
Liu, Y., Qiao, F. & Wang, H. Enhanced Proteostasis in Post-ischemic Stroke Mouse Brains by Ubiquilin-1 Promotes Functional Recovery. Cell Mol Neurobiol 37, 1325–1329 (2017). https://doi.org/10.1007/s10571-016-0451-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-016-0451-3