title: Heat shock protein 60: an endogenous inducer of dopaminergic cell death in Parkinson disease.
abstract: BACKGROUND
  Increasing evidence suggests that inflammation associated with microglial cell activation in the substantia nigra (SN) of patients with Parkinson disease (PD) is not only a consequence of neuronal degeneration, but may actively sustain dopaminergic (DA) cell loss over time. We aimed to study whether the intracellular chaperone heat shock protein 60 (Hsp60) could serve as a signal of CNS injury for activation of microglial cells.
  
  
  METHODS
  Hsp60 mRNA expression in the mesencephalon and the striatum of C57/BL6 mice treated with MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and the Hsp60/TH mRNA ratios in the SN of PD patients and aged-matched subjects were measured. To further investigate a possible link between the neuronal Hsp60 response and PD-related cellular stress, Hsp60 immunoblot analysis and quantification in cell lysates from SH-SY5Y after treatment with 100 μM MPP+ (1-methyl-4-phenylpyridinium) at different time points (6, 12, 24 and 48 hours) compared to control cells were performed. Additional MTT and LDH assay were used. We next addressed the question as to whether Hsp60 influences the survival of TH+ neurons in mesencephalic neuron-glia cultures treated either with MPP+ (1 μM), hHsp60 (10 μg/ml) or a combination of both. Finally, we measured IL-1β, IL-6, TNF-α and NO-release by ELISA in primary microglial cell cultures following treatment with different hHsp60 preparations. Control cultures were exposed to LPS.
  
  
  RESULTS
  In the mesencephalon and striatum of mice treated with MPTP and also in the SN of PD patients, we found that Hsp60 mRNA was up-regulated. MPP+, the active metabolite of MPTP, also caused an increased expression and release of Hsp60 in the human dopaminergic cell line SH-SY5Y. Interestingly, in addition to being toxic to DA neurons in primary mesencephalic cultures, exogenous Hsp60 aggravated the effects of MPP+. Yet, although we demonstrated that Hsp60 specifically binds to microglial cells, it failed to stimulate the production of pro-inflammatory cytokines or NO by these cells.
  
  
  CONCLUSIONS
  Overall, our data suggest that Hsp60 is likely to participate in DA cell death in PD but via a mechanism unrelated to cytokine release.
authors.forename: Carmen
authors.initials: C
authors.lastname: Noelker
authors.forename: Lydie
authors.initials: L
authors.lastname: Morel
authors.forename: Anke
authors.initials: A
authors.lastname: Osterloh
authors.forename: Daniel
authors.initials: D
authors.lastname: Alvarez-Fischer
authors.forename: Thomas
authors.initials: T
authors.lastname: Lescot
authors.forename: Minka
authors.initials: M
authors.lastname: Breloer
authors.forename: Maike
authors.initials: M
authors.lastname: Gold
authors.forename: Wolfgang H
authors.initials: WH
authors.lastname: Oertel
authors.forename: Carmen
authors.initials: C
authors.lastname: Henze
authors.forename: Patrick P
authors.initials: PP
authors.lastname: Michel
authors.forename: Richard C
authors.initials: RC
authors.lastname: Dodel
authors.forename: Lixia
authors.initials: L
authors.lastname: Lu
authors.forename: Etienne C
authors.initials: EC
authors.lastname: Hirsch
authors.affiliation: CR-ICM, INSERM UMR_S1127, Université Pierre et Marie Curie Paris 06 UMR_S1127, CNRS UMR 7225, Groupe Hospitalier Pitié-Salpêtrière, 75013 Paris, France. stephane.hunot@upmc.fr.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Hunot
authors.forename: Andreas
authors.initials: A
authors.lastname: Hartmann
mesh_terms: D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000494:pharmacology; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism*; D003342:Corpus Striatum / Q000473:pathology*; D004195:Disease Models, Animal; D015259:Dopamine Agents / Q000494:pharmacology; D059290:Dopaminergic Neurons / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D020267:MPTP Poisoning / Q000473:pathology*; D008297:Male; D008636:Mesencephalon / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D018834:Chaperonin 60; D015259:Dopamine Agents; D012333:RNA, Messenger; D009569:Nitric Oxide; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D007770:L-Lactate Dehydrogenase; D014446:Tyrosine 3-Monooxygenase