Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: Effect of exogenous IL6 on caloric intake and body mass. Wild-type mice were fed on chow or high-fat diet (HFD) and treated intraperitoneally (IP) with exogenous IL6 or saline as depicted in A. Body mass ( B , D ) and caloric intake ( C , E ) were determined in mice fed chow ( B , C ) and HFD ( D , E ). In all experiments, n = 5 mice/group. Data was analyzed using two-way ANOVA with repeated measures and Bonferroni post-test. ** p < 0.01; *** p < 0.001

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: Hypothalamic transcript expression of IL6. In order to identify obese-prone ( H ) and obese-resistant ( L ) mice, we performed the procedure as depicted in A. The expression of hypothalamic IL6 was determined using real-time PCR ( B ). Palmitate was injected in a single intracerebroventricular (ICV) dose and hypothalamus was collected ( C ) for determinations of IL6 ( D ), TNFα ( E ), and IL1β ( F ) transcripts. B , n = 7–8 mice/group; D – F , n = 5 mice/group; * p < 0.05, ** p < 0.01, *** p < 0.001. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Injection, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 effect on hypothalamic cell proliferation and survival. Mice were treated with IL6 and BrdU according to the protocols as depicted in A and D . The mediobasal hypothalamus was prepared for analysis using immunofluorescence and confocal microscopy; cell counting was performed using 20× and 40× magnifications. Representative images are presented in B (proliferation assay) and E (survival assay). Total BrdU countings are presented in C (proliferation assay) and F (survival assay). In both assays, n = 6 mice/group. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Immunofluorescence, Confocal Microscopy, Cell Counting, Proliferation Assay, Clonogenic Cell Survival Assay, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 induces neurogenesis in the hypothalamus. Mice were treated with a single intraperitoneal dose of IL6, according to the protocol as depicted in A . The hypothalamic expressions of Sox6 ( B ) and Sox2 ( C ) were determined using real-time PCR. The transcript expressions of Sox6 ( D ) and doublecortin ( E ) were determined in the hypothalami collected from obese-prone ( H ) and obese-resistant ( L ) mice and treated with a single dose of IL6. F Mice were treated with IL6 and BrdU according to the protocols as depicted in Fig. . The mediobasal hypothalamus was prepared for analysis using immunofluorescence and confocal microscopy; cell counting was performed using 20× and 40× magnifications. Total BrdU/NeuN counting is depicted in G (neurogenesis assay). B , C n = 5 mice/group; D , E n = 8–10 mice/group; F , G , n = 5 mice/group. * p < 0.05, ** p < 0.01. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Real-time Polymerase Chain Reaction, Immunofluorescence, Confocal Microscopy, Cell Counting, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: Analysis of single-cell RNA sequencing data reveal the expression pattern of IL6-receptor encoding genes in the arcuate nucleus of the hypothalamus. A Diffusion-based manifold approximation and projection (dbMAP) embedding of 20,921 single-cell transcriptomes of the arcuate nucleus of the hypothalamus, colored by their annotated cluster of origin. In this embedding, similar cells are mapped tightly, whereas dissimilar cells are mapped apart from each other, rendering clusters of cell types. Cells were annotated by their main cell type as defined in the original study. B (top) Il6ra mRNA expression, key-colored by expression intensity; Il6ra is found mainly in perivascular macrophages/microglia and ependymocytes and also in astrocytes and endothelial cells. B (bottom) Cx3cr1 (a canonical microglial marker) mRNA expression, key-colored by expression intensity. C (top) Il6st mRNA expression, key-colored by expression intensity; Il6st is broadly expressed in the arcuate with some overlapping with the expression pattern of Ntrk2 (bottom), which encodes a neurotrophic receptor. D Violin plot of the mRNA expressions of Il6, Il6ra, Il6st, Cx3cr1, Vim, Ntrk2, and Gfap in the arcuate hypothalamus, per main cell types. Oligodend, oligodendrocyte; NG2/OPC, oligodendrocyte progenitor cells; PVMMicro, perivascular macrophages/microglia; VLMCs, vascular and leptomeningeal cells; ParsTuber, pars tuberalis

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: RNA Sequencing Assay, Expressing, Diffusion-based Assay, Marker

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 deficiency predisposes to obesity. IL6 was determined in the serum of wild-type (WT) and IL6 knockout (KO) mice treated or not with 1.0 ng exogenous IL6 ( A ). Body mass was determined at age 8 weeks ( B ). The expression of transcripts encoding for hypothalamic neurotransmitters ( C , D ) and neurogenesis-related genes ( E , F ) were determined using real-time PCR in hypothalamic samples collected in mice aged 7 days ( C , E ) or 8 weeks ( D , F ). A n = 3–4 mice/group; n = 25 mice/group; C , E n = 5–7 mice/group; D , F n = 9 mice/group. * p < 0.05, *** p < 0.001. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Knock-Out, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Interleukin-6 actions in the hypothalamus protects against obesity and is involved in the regulation of neurogenesis

doi: 10.1186/s12974-021-02242-8

Figure Lengend Snippet: IL6 induce neurogenic factors expression in hypothalamic neuroprogenitor cells (NPCs). Transcripts were determined using real-time PCR in the NPCs of WT and IL6 KO mice after a 5-day differentiation period: first, WT and IL6 KO hypothalamic cells were analyzed in basal conditions ( A ), and then, IL6 KO NPCs were treated with exogenous IL6 in doses depicted ( B ). Representative immunocytochemistry images of 7-day differentiated cells (green, DCX; red, GFAP; blue, DAPI) ( C ) and DCX-positive cell counting ( D ), and representative immunocytochemistry images of 14-day differentiated cells (green, MAP2; red, GFAP; blue, DAPI) ( E ) and MAP2-positive cell counting ( F ). After 10- ( G , I , K – M ) or 14-day ( H and J ) differentiation period, transcripts were determined using real-time PCR in the NPCs of WT and IL6 KO mice treated with exogenous IL6 in doses depicted in the panels (G-M). A , B n = 12 (3–4 well/group, 2 independent experiments); C – F n = 24 (4 well/group, 2 independent experiments); G – M n = 48 (6–9 well/group, 3 independent experiments). * p < 0.05, ** p < 0.01. In bar graphs, results are presented as mean ± standard deviation

Article Snippet: For that, we determined body mass and blood IL-6 levels at baseline conditions and 4 h after treatment with recombinant IL6 (Sigma) (1 ng ip) and hypothalamic gene expression analysis in baseline conditions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunocytochemistry, Cell Counting, Standard Deviation