Prolonging the integrated stress response enhances CNS remyelination in an inflammatory environment

  1. Yanan Chen
  2. Rejani B Kunjamma
  3. Molly Weiner
  4. Jonah R Chan
  5. Brian Popko  Is a corresponding author
  1. Department of Neurology, Division of Multiple Sclerosis and Neuroimmunology, Northwestern University Feinberg School of Medicine, United States
  2. Weill Institute for Neuroscience, Department of Neurology, University of California, San Francisco, United States
8 figures, 1 table and 1 additional file

Figures

Sephin1 treatment enhances remyelination after inflammatory demyelination.

(A) Cumulative clinical scores of C57BL/6J female mice immunized with MOG35-55/CFA to induce chronic EAE, treated with vehicle (n = 7) and 8 mg/kg Sephin1 (n = 7) from the peak disease. *p<0.05. …

Figure 1—source data 1

Disease scores and axon measurement of late stage of EAE.

https://cdn.elifesciences.org/articles/65469/elife-65469-fig1-data1-v1.xlsx
Figure 2 with 1 supplement
Schematics of GFAP-tTA;TRE-IFN-γ double-transgenic mouse model of cuprizone demyelination/remyelination.

(A) GFAP-tTA mice are mated with TRE-IFN-γ mice to produce double-positive animals. When these mice are maintained on doxycycline (Dox), the expression of the IFN-γ is repressed. When they are …

Figure 2—figure supplement 1
GFAP-tTA;TRE-IFN-γ mice express IFN-γ after release from doxycycline.

Real-time qPCR analyses for detection of mRNA levels of ectopically expressed IFN-γ in the brains of GFAP-tTA;TRE-IFN-γ mice. (A) The expression of IFN-γ in the GFAP-tTA;TRE-IFN-γ/GADD34 KO or WT …

GADD34 deficiency protects remyelinating oligodendrocytes in the presence of IFN-γ.

The corpora callosa of GFAP-tTA;TRE-IFN-γ/GADD34 KO or WT were taken at W0, W5, and W8. (A) Immunofluorescent staining for ASPA (a mature oligodendrocyte marker) and DAPI (nuclei). Scale bar = 100 …

GADD34 deficiency enhances remyelination in the presence of IFN-γ.

The corpora callosa of GFAP-tTA;TRE-IFN-γ/GADD34 KO or WT were harvested for EM processing. (A) Representative EM images of axons in the corpus callosum at W0 and W5. Scale bar = 1 µm. (B) …

Sephin1 protects remyelinating oligodendrocytes in the presence of IFN-γ.

The corpora callosa of GFAP-tTA;TRE-IFN-γ were taken at W0 and W3 prior to any treatment as well as after either vehicle or Sephin1 treatment at W5 and W8. (A) Immunofluorescent staining for ASPA (a …

Figure 5—source data 1

The number of ASAP+ cells in Veh and Seph groups.

https://cdn.elifesciences.org/articles/65469/elife-65469-fig5-data1-v1.xlsx
Sephin1 enhances remyelination in the presence of IFN-γ.

The corpora callosa of GFAP-tTA;TRE-IFN-γ were harvested for EM processing. (A) Representative EM images of axons in the corpus callosum at W5. Scale bar = 1 µm. (B) Representative EM images of …

Figure 6—source data 1

The number of myelinated axons in Veh and Seph groups.

https://cdn.elifesciences.org/articles/65469/elife-65469-fig6-data1-v1.xlsx
Figure 7 with 2 supplements
GADD34 deficiency or Sephin1 does not affect OPC proliferation during remyelination.

(A) Immunofluorescent staining for PDGFRα (an OPC marker), Ki67 and DAPI (nuclei) from the corpus callosum of GFAP-tTA;TRE-IFN-γ/GADD34 KO or WT was taken at W5 and W8. Scale bar = 50 µm. …

Figure 7—source data 1

The number of total OPCs and proliferating OPCs.

https://cdn.elifesciences.org/articles/65469/elife-65469-fig7-data1-v1.xlsx
Figure 7—figure supplement 1
GADD34 deficiency does not affect microglial activation during remyelination in the presence of IFN-γ.

The corpus callosum of GFAP-tTA;TRE-IFN-γ/GADD34 KO or WT was taken at W0, W5 and W8. (A) Immunofluorescent staining for IBA1 (a microglia marker), MBP (a myelin marker) and DAPI. Scale bar = 50 µm. …

Figure 7—figure supplement 2
Sephin1 treatment does not affect microglial activation during remyelination in the presence of IFN-γ.

The corpus callosum of GFAP-tTA;TRE-IFN-γ was taken at W0, W5, and W8 with vehicle or Sephin1 treatment. (A) Immunofluorescent staining for IBA1, MBP, and DAPI. Scale bar = 50 µm. (B) Quantification …

Combined treatment of Sephin1 and BZA accelerates remyelination.

GFAP-tTA;TRE-IFN-γ mice were released from Dox at W0 and given cuprizone chow for 5 weeks. Treatment with vehicle, Sephin1, BZA or combined BZA and Sephin1 was started at W3, and the corpus callosum …

Figure 8—source data 1

The number of ASAP+ cells, myelinated axons and g-ratio of various treatments.

https://cdn.elifesciences.org/articles/65469/elife-65469-fig8-data1-v1.xlsx

Tables

Key resources table
Reagent type
(species) or
resource
DesignationSource or
reference
IdentifiersAdditional
information
Strain, strain background (M. musculus)C57Bl/6JJackson labRRID:IMSR_JAX:000664
Strain, strain background (M. musculus)GFAP-tTA miceLin et al., 2004RRID:IMSR_JAX:005964
Strain, strain background (M. musculus)TRE-IFN-γ miceLin et al., 2004RRID:IMSR_JAX:009344
Strain, strain background (M. musculus)Ppp1r15a-/-(GADD34 KO) miceGift from David RonRRID:MGI:3040935
Chemical compound, drugdoxycyclineSigma-AldrichCat # D9891
Chemical compound, drug0.2% cuprizoneEnvigoCat # TD.160049
Chemical compound,
drug
Sephin1ApexbioCat # A8708
Chemical compound, drugbazedoxifene acetateSigma-AldrichCat # PZ0018
Chemical compound, drugMOG 35-55peptideGenemed synthesisCat # MOG3555-P-5
Chemical compound, drugpertussis toxinList Biological LaboratoriesCat # 179
Sequence-based reagentGapdh-fThis paperqPCR primerTGTGTCCGTCGTGGATCTGA
Sequence-based reagentGapdh-rThis paperqPCR primerTTGCTGTTGAAGTCGCAGGAG
Sequence-based reagentIfng-fThis paperqPCR primerGATATCTGGAGGAACTGGCAAAA
Sequence-based reagentIfng-rThis paperqPCR primerCTTCAAAGAGTCTGAGGTAGAAAGAGATAAT
AntibodyAnti-MBP (mouse monoclonal)AbcamCat # ab24567
RRID:AB_448144
(1:700)
AntibodyAnti-ASPA (rabbit polyclonal)GenetexCat # GTX113389
RRID:AB_2036283
(1:500)
AntibodyAnti-Ki67 (rabbit polyclonal)AbcamCat # AB15580
RRID:AB_443209
(1:100)
AntibodyAnti-PDGFR-alpha (mouse monoclonal)BD BiosciencesCat # 558774
RRID:AB_397117
(1:100)
AntibodyAnti-Iba1 (rabbit polyclonal)Wako Pure ChemicalCat # 019–19741
RRID:AB_839504
(1:500)
Software, algorithmImageJNational Institutes of HealthRRID:SCR_003070
Software, algorithmPrism 6.0GraphpadRRID:SCR_002798

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