Lactate receptor HCAR1 regulates neurogenesis and microglia activation after neonatal hypoxia-ischemia
- Department of Microbiology, Oslo University Hospital and University of Oslo, Norway
- Division of Physiology, Institute of Basic Medical Sciences, University of Oslo, Norway
- Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Norway
- The Brain and Muscle Energy Group, Institute of Oral Biology, Faculty of Dentistry, University of Oslo, Norway
- Center for Healthy Aging, Department of Neuroscience and Pharmacology, Faculty of Health Sciences, University of Copenhagen, Denmark
Figures
Figure 1
HCAR1 does not protect the brain from acute tissue damage following cerebral HI, but promotes brain tissue regeneration.
(A) Representative images of TTC-stained brain sections from WT and HCAR1 KO mice 24 hr after HI. TTC turns red upon reacting with mitochondrial respiratory enzymes in viable tissue, whereas …
Figure 2 with 1 supplement
HCAR1 regulates neural progenitor cell proliferation.
A-I Neurosphere formation from HCAR1 KO and WT cells. (A–B) Images of neurospheres from WT (A) and HCAR1 KO (B) mice. (C–F) Fluorescence images from WT (C, E) and HCAR1 KO (D, F) dissociated …
Figure 2—figure supplement 1
Density data from subventricular-intermediate zone (SVZ/IZ) and quantification proliferated cells and neuroblasts in the ventricular-subventricular zone (VZ/SVZ) after HI.
(A) Density of BrdU + cells within the SVZ/IZ. WT contra vs ipsi, p<0.001, df 11, n=6. KO contra vs ipsi, p=0.95, df 11, n=7. (B) SVZ/IZ density of DCX + cells. WT contra vs ipsi, p<0.001, df 15, …
Figure 3 with 1 supplement
HCAR1 KO mice have deficient activation and proliferation of microglia after HI.
(A-D) Confocal images from the peri-infarct zone (b, d, indicated as square in cartoon) and corresponding contralateral area (A, C) of coronal mouse brain sections from WT (A–B) and KO (C–D) …
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Figure 3—source code 1
BrdU WEKA.
Algorithms used for WEKA segmentation for BrdU-staining.
- https://cdn.elifesciences.org/articles/76451/elife-76451-fig3-code1-v1.zip
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Figure 3—source code 2
DAPI WEKA.
Algorithms used for WEKA segmentation for DAPI-staining.
- https://cdn.elifesciences.org/articles/76451/elife-76451-fig3-code2-v1.zip
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Figure 3—source code 3
IBA1 WEKA.
Algorithms used for WEKA segmentation for IBA1-staining.
- https://cdn.elifesciences.org/articles/76451/elife-76451-fig3-code3-v1.zip
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Figure 3—source code 4
Script Microglia analysis.
Script for analysis of immunostaining of microglia.
- https://cdn.elifesciences.org/articles/76451/elife-76451-fig3-code4-v1.zip
Figure 3—figure supplement 1
Density measurements of microglia in the peri-infarct zone.
(A) Density of microglia (IBA1 + cells/mm2) in the peri-infarct zone. WT contra vs ipsi p=0.002, df 16, n=8. KO contra vs ipsi p=0.07, df 16, n=10. (B) Density of proliferating microglia (i.e. cells …
Figure 4 with 1 supplement
Astrocyte and oligodendrocyte proliferation after HI.
(A-D) Confocal images showing immunolabelling of GFAP +Astrocytes (green) and proliferated BrdU + cells (magenta) in striatal peri-infarct area from contralateral (control, ctrl) and ipsilateral …
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Figure 4—source code 1
Script GFAP analysis.
Script for analysis of immunostaining of GFAP + cells.
- https://cdn.elifesciences.org/articles/76451/elife-76451-fig4-code1-v1.zip
Figure 4—figure supplement 1
Density measurements of GFAP, Olig2, and APC in the peri-infarct zone.
(A) Density of peri-infarct zone (striatal) GFAP +astrocytes. WT contra vs ipsi, p<0.001, df 18, n=12. KO contra vs ipsi, p=0.002, df 18, n=8. (B) Proliferated astrocytes. WT contra vs ipsi, …
Figure 5 with 4 supplements
HCAR1 regulates transcriptional response to ischemia including cell cycle and complement pathway.
(A) PCA plot of transcriptome data from subventricular zone tissue samples from the ipsilateral (HI-damaged) and contralateral (control, ctrl) hemisphere in HCAR1 KO and WT mice. Each point …
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Figure 5—source data 1
Raw images of western blots in Figure 5.
- https://cdn.elifesciences.org/articles/76451/elife-76451-fig5-data1-v1.zip
Figure 5—figure supplement 1
Gene set enrichment analysis (GSEA) of subventricular zone from HCAR1 KO ipsi versus WT ipsi.
The 20 most up- or downregulated pathways are included.
Figure 5—figure supplement 2
Gene set enrichment analysis (GSEA) of subventricular zone from WT ipsi (HI) versus WT contra (control).
The 20 most up- or downregulated pathways are included.
Figure 5—figure supplement 3
Gene set enrichment analysis (GSEA) of subventricular zone from HCAR1 KO contra versus WT contra.
The 20 most up- or downregulated pathways are included. Note that only one pathway has FDR ≤0.05.
Figure 5—figure supplement 4
Gene set enrichment analysis (GSEA) of subventricular zone from HCAR1 KO ipsi (HI) versus HCAR1 KO contra (control).
The 20 most up- or downregulated pathways are included.
Figure 6
Proposed model for the role of HCAR1 in neonatal HI.
During and after HI, the extracellular concentration of lactate ([lac]o) is elevated. Top panel: In WT mice, the elevated lactate causes HCAR1 activation, which induces transcription of genes …
Additional files
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Supplementary file 1
Number of differentially expressed genes (DEGs) between the different experimental groups.
Thresholds ->logFC ≥ 0.5 (UP DEGs) and ≤–0.5 (DOWN DEGs); all of them significant at adjusted P-value <0.05. SVZ, subventricular zone; hc, hippocampus.
- https://cdn.elifesciences.org/articles/76451/elife-76451-supp1-v1.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/76451/elife-76451-transrepform1-v1.pdf
