Journal: Nature Communications

Article Title: Differential IL18 signaling via IL18 receptor and Na-Cl co-transporter discriminating thermogenesis and glucose metabolism regulation

doi: 10.1038/s41467-022-35256-8

Figure Lengend Snippet: a – d Bodyweight gain, glucose tolerance test (GTT), AUC of GTT, insulin tolerance test (ITT), AUC of ITT (WT: n = 10; Ncc −/− : n = 11, Il18r −/− : n = 11; Il18r −/− Ncc −/− : n = 11) ( a ), EAT, SAT, BAT, and liver weights (WT: n = 9; Ncc −/− : n = 14, Il18r −/− : n = 13; Il18r −/− Ncc −/− : n = 11) ( b ), energy intake (WT: n = 12; Ncc −/− : n = 11, Il18r −/− : n = 10; Il18r −/− Ncc −/− : n = 10) ( c ), and plasma insulin (WT: n = 11; Ncc −/− : n = 10, Il18r −/− : n = 13; Il18r −/− Ncc −/− : n = 8) and leptin (WT: n = 12; Ncc −/− : n = 15, Il18r −/− : n = 10; Il18r −/− Ncc −/− : n = 14) levels ( d ) from HFD-fed WT, Ncc −/− , Il18r −/− , and Il18r −/− Ncc −/− mice for 12 weeks. e – g Mouse metabolic parameters, including oxygen consumption (VO 2 ) ( e ), carbon dioxide production (VCO 2 ) ( f ), and energy expenditure ( g ) and their average values from full day cycle, light cycle, and dark cycle during 48 hrs of monitoring in WT, Ncc −/− , Il18r −/− , and Il18r −/− Ncc −/− mice (WT: n = 5; Ncc −/− : n = 6, Il18r −/− : n = 6; Il18r −/− Ncc −/− : n = 6). h Representative images of hematoxylin and eosin (H&E) staining and adipocyte sizes in EAT, SAT, and BAT from indicated mouse groups (WT: n = 14; Ncc −/− : n = 15, Il18r −/− : n = 9; Il18r −/− Ncc −/− : n = 12); scale bar: 50 μm. i . Plasma IL1β (WT: n = 6; Ncc −/− : n = 4, Il18r −/− : n = 4; Il18r −/− Ncc −/− : n = 7), IL6 (WT: n = 9; Ncc −/− : n = 12, Il18r −/− : n = 7; Il18r −/− Ncc −/− : n = 9), MCP1 (WT: n = 8; Ncc −/− : n = 12, Il18r −/− : n = 13; Il18r −/− Ncc −/− : n = 9), IFN-γ (WT: n = 10; Ncc −/− : n = 16, Il18r −/− : n = 12; Il18r −/− Ncc −/− : n = 14), and TNF-α (WT: n = 6; Ncc −/− : n = 4, Il18r −/− : n = 6; Il18r −/− Ncc −/− : n = 4) levels from indicated mouse groups. j Mac-3 immunostaining of representative EAT sections and quantification from indicated mouse groups (WT: n = 11; Ncc −/− : n = 12, Il18r −/− : n = 17; Il18r −/− Ncc −/− : n = 12); scale: 50 μm. Data are mean ± SEM, two-way ANOVA repeated-measures ( a ) or one-way ANOVA test ( b-j ), followed by LSD post-test. Sample sizes were all biologically independent samples.

Article Snippet: The plasma levels of IL18 (BMS618-3, Invitrogen), insulin (90080, Crystal Chem, Elk Grove Village, IL), leptin (RD291001200R, BioVendor, Asheville, NC), IL1β (88-7013-22), TNF-α (88-7324-22), IL6 (88-7064-88), MCP1 (88-7391-88), IFN-γ (88-7314-88) from Invitrogen were assessed using the mouse ELISA kits according to the manufacturers.

Techniques: Staining, Immunostaining

Journal: Nutrients

Article Title: Stimulation of GHRH Neuron Axon Growth by Leptin and Impact of Nutrition during Suckling in Mice

doi: 10.3390/nu15051077

Figure Lengend Snippet: Primary and secondary antibodies used for immunohistochemistry experiments.

Article Snippet: Plasma leptin concentrations in 10-day-old male mice were determined individually with the Mouse/Rat Leptin (R&D systems) ELISA kit, according to the manufacturer’s instructions.

Techniques: Immunohistochemistry

Journal: Nutrients

Article Title: Stimulation of GHRH Neuron Axon Growth by Leptin and Impact of Nutrition during Suckling in Mice

doi: 10.3390/nu15051077

Figure Lengend Snippet: Underfeeding during suckling results in lower body weight and plasma leptin levels. ( A ) Increases in litter size from six (normally fed, open bars) to nine (underfed, blue bars) pups per dam are associated with a lower body weight in pups, by the age of seven days ( n = 32 normally fed pups and 41 underfed pups of both sexes). ( B ) Underfeeding during suckling was associated with lower plasma levels of leptin at 10 days of age, as determined by ELISA ( n = 8 per group). ( C ) Illustration (40X magnification obtained with a BX43 Olympus fluorescence microscope equipped with a DP73 CCD camera) of a growing axon from an arcuate nucleus explant cultured in vitro, showing that the GHRH+ axon in green (uppermost image) expresses the leptin receptor in red (middle image). A merged image is shown at the bottom (arrow). Scale bars represent 20 µm. Data are presented as the mean ± SEM. Comparisons were performed in Mann–Whitney tests, with ** p < 0.01 and *** p < 0.001.

Article Snippet: Plasma leptin concentrations in 10-day-old male mice were determined individually with the Mouse/Rat Leptin (R&D systems) ELISA kit, according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Fluorescence, Microscopy, Cell Culture, In Vitro, MANN-WHITNEY

Journal: Nutrients

Article Title: Stimulation of GHRH Neuron Axon Growth by Leptin and Impact of Nutrition during Suckling in Mice

doi: 10.3390/nu15051077

Figure Lengend Snippet: Leptin stimulates axon growth in GHRH neurons in arcuate nucleus explants from normally fed pups. ( A ) Illustrative IHC of AgRP neurons from arcuate nucleus explants micro-dissected from one-week-old normally fed pups, in control conditions (left panels), and after stimulation with leptin (middle panels) and with leptin/IGF-1 (right panels). ( B ) Illustrative images of dual IHC for the axons of total arcuate nucleus neurons labeled with NF (top panels in red) and GHRH neurons labeled with eGFP (bottom panels in green), in the same conditions. Scale bars represent 1000 µm for AgRP+ and NF+ IHC (4X magnification) and 200 µm for GHRH+ IHC (10X magnification), for images from a BX612 Olympus fluorescence microscope equipped with a DP71 CCD camera. ( C ) Quantification of the growth of AgRP axons after 24 h of stimulation with leptin or leptin/IGF-I relative to control conditions ( n = 4 experiments), and ( D ) of the growth of NF (plain bars) and GHRH (dashed bars) axons ( n = 5–7 experiments). Data are presented as the mean ± SEM. Results were compared in a one-way ANOVA with the Newman–Keuls post hoc test (c) or a two-way ANOVA with Bonferroni correction (d), with *: p < 0.05 and **: p < 0.01 vs. control conditions and #: p < 0.05 vs. leptin stimulation.

Article Snippet: Plasma leptin concentrations in 10-day-old male mice were determined individually with the Mouse/Rat Leptin (R&D systems) ELISA kit, according to the manufacturer’s instructions.

Techniques: Control, Labeling, Fluorescence, Microscopy

Journal: Nutrients

Article Title: Stimulation of GHRH Neuron Axon Growth by Leptin and Impact of Nutrition during Suckling in Mice

doi: 10.3390/nu15051077

Figure Lengend Snippet: Signaling pathways involved in the axon growth of arcuate neurons in explants from normally fed pups. ( A ) Illustrative triple IHC of arcuate nucleus explants from the hypothalamus micro-dissected from one-week-old normally fed pups under control conditions (first panel), and following stimulation with leptin (second panel), leptin/NSC (third panel), leptin/LY (fourth panel) and leptin/PD (fifth panel), with NF (top panels in green), GHRH (middle panels in red) and AgRP (bottom panels in blue). Scale bars represent 100 µm for NF+ IHC (4X magnification) and 200 µm for GHRH+ and AgRP+ IHC (10X magnification), on images obtained with an Olympus BX43 fluorescence microscope equipped with a DP73 CCD camera. Quantifications of the growth of ( B ) NF ( n = 5–9 experiments), ( C ) GHRH ( n = 4–10 experiments) and ( D ) AgRP ( n = 5–10 experiments) axons stimulated for 24 h with leptin alone, or in combination with one of the three inhibitors (NSC_33994, LY_294002 or PD_0325901). Data are presented as the mean ± SEM. Results were analyzed by two-way ANOVA with Bonferroni correction, with *: p < 0.05 and ***: p < 0.001 vs. control conditions and ###: p < 0.001 vs. leptin stimulation.

Article Snippet: Plasma leptin concentrations in 10-day-old male mice were determined individually with the Mouse/Rat Leptin (R&D systems) ELISA kit, according to the manufacturer’s instructions.

Techniques: Protein-Protein interactions, Control, Fluorescence, Microscopy

Journal: Nutrients

Article Title: Stimulation of GHRH Neuron Axon Growth by Leptin and Impact of Nutrition during Suckling in Mice

doi: 10.3390/nu15051077

Figure Lengend Snippet: GHRH neurons from underfed pups are resistant to leptin for the stimulation of axon growth. ( A ) Illustrative IHC of arcuate nucleus explants from the hypothalamus micro-dissected from one-week-old underfed pups in control conditions (left panel), and after stimulation with leptin alone (middle panel), or with leptin/IGF-1 (right panel), with AgRP axons labeled in orange. ( B ) Axons from total arcuate nucleus neurons and GHRH neurons labeled by dual-IHC for neurofilament (NF, in red) and eGFP (in green), respectively. Scale bars represent 1000 µm for AgRP+ and NF+ IHC (4X magnification) and 200 µm for GHRH+ IHC (10X magnification), on images from an Olympus BX612 fluorescence microscope equipped with a DP71 CCD camera. ( C ) Quantification of the growth of AgRP axons stimulated by incubation for 24 h with leptin or leptin/IGF-I, relative to control conditions ( n = 5 experiments), and ( D ) quantification of the growth of NF (plain bars) and GHRH (dashed bars) axons ( n = 6 experiments). Data are presented as the mean ± SEM. Results were compared in a one-way ANOVA with the Newman–Keuls post hoc test (c) or in a two-way ANOVA with Bonferroni correction (d), with *: p < 0.05.

Article Snippet: Plasma leptin concentrations in 10-day-old male mice were determined individually with the Mouse/Rat Leptin (R&D systems) ELISA kit, according to the manufacturer’s instructions.

Techniques: Control, Labeling, Fluorescence, Microscopy, Incubation

Journal: Nutrients

Article Title: Stimulation of GHRH Neuron Axon Growth by Leptin and Impact of Nutrition during Suckling in Mice

doi: 10.3390/nu15051077

Figure Lengend Snippet: Alterations to leptin-stimulated signaling pathways in arcuate nucleus explants from underfed pups. The activation of the three signaling pathways by the exposure of seven-day explants to leptin (+) for 15 min was different in explants from underfed pups (blue bars) and in explants from normally fed pups (open bars). Data are presented as the fold induction of phosphorylated protein/total protein ratios (normalized against actin) for stimulated relative to unstimulated conditions for ( A ) phosphorylated Jak2/Jak2/actin ( n = 5 per group), ( B ) phosphorylated Stat3/Stat3/actin ( n = 5–7 per group), ( C ) phosphorylated-AKT/AKT/actin ( n = 9 per group), ( D ) phosphorylated MEK1/MEK1/actin ( n = 5–6 per group), ( E ) phosphorylated ERK1/ERK1/actin (left panel) and phosphorylated-ERK2/ERK2/actin (right panel; n = 8 per group). Data are presented as the mean ± SEM, with a Mann–Whitney analysis, with *: p < 0.05, **: p < 0.01.

Article Snippet: Plasma leptin concentrations in 10-day-old male mice were determined individually with the Mouse/Rat Leptin (R&D systems) ELISA kit, according to the manufacturer’s instructions.

Techniques: Protein-Protein interactions, Activation Assay, MANN-WHITNEY

Journal: PLoS ONE

Article Title: A Predictive Model of the Dynamics of Body Weight and Food Intake in Rats Submitted to Caloric Restrictions

doi: 10.1371/journal.pone.0100073

Figure Lengend Snippet: Plasma hormones and glucose assays.

Article Snippet: Plasma ghrelin and leptin assays were performed using immunoassays (acylated rat/mouse ghrelin #A05117 and rat/mouse leptin EIA #A05176, Cayman, SpiBio, Montigny le Bretonneux, France) according to the manufacturer’s recommendations.

Techniques: Clinical Proteomics

Journal: PLoS ONE

Article Title: A Predictive Model of the Dynamics of Body Weight and Food Intake in Rats Submitted to Caloric Restrictions

doi: 10.1371/journal.pone.0100073

Figure Lengend Snippet: Model variables.

Article Snippet: Plasma ghrelin and leptin assays were performed using immunoassays (acylated rat/mouse ghrelin #A05117 and rat/mouse leptin EIA #A05176, Cayman, SpiBio, Montigny le Bretonneux, France) according to the manufacturer’s recommendations.

Techniques: Clinical Proteomics