Journal: Frontiers in cell and developmental biology

Article Title: Increased adipose tissue lymphatic vessel density inhibits thermogenesis through elevated neurotensin levels.

doi: 10.3389/fcell.2023.1100788

Figure Lengend Snippet: FIGURE 2 Lymphangiogenesis in AdipoVD mice result in increased neurotensin production. (A) Neurotensin and neurotensin receptor relative mRNA expression in BAT of AdipoVD mice compared to Controls. (B) Neurotensin and neurotensin receptor relative mRNA expression in SQAT of AdipoVD mice compared to Controls. (C) ELISA quantification of neurotensin concentrations from BAT normalized to tissue protein levels. (D) ELISA quantification of neurotensin concentrations from SQAT normalized to tissue protein levels. (E) Immunofluorescence of neurotensin (green; Nts) and lymphatics (red; LYVE-1) in the BAT depot from Control and AdipoVD mice on HFD. (A–E) n = 10 control/n = 7 AdipoVD. Bars = 50 µm.

Article Snippet: Neurotensin protein levels from tissue lysates were measured by Neurotensin EIA kit (RayBiotech) according to manufacturer’s protocols and normalized to the total protein concentration.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in cell and developmental biology

Article Title: Increased adipose tissue lymphatic vessel density inhibits thermogenesis through elevated neurotensin levels.

doi: 10.3389/fcell.2023.1100788

Figure Lengend Snippet: FIGURE 3 Temperature dysfunction in AdipoVD mice as a result of increased neurotensin levels. (A) Body temperature of Control and AdipoVD mice during acute under cold exposure at 4°C. (B) Browning and thermogenesis-associated gene expression in AdipoVD BAT relative to Controls under acute cold exposure. (C) Immunofluorescence of potential thermogenesis (green; UCP-1) and lymphatic (red; LYVE-1) from BAT depot of Control and AdipoVD following cold exposure. (C) Immunofluorescence of UCP-1 (green) and lymphatics (red; LYVE-1) from BAT depot of Control and AdipoVD following cold exposure. (D) Browning and thermogenesis-associated gene expression in AdipoVD SQAT relative to Controls following cold exposure. (E) Immunofluorescence of potential thermogenesis (green; UCP-1) and lymphatic (red; LYVE-1) from SQAT depot of Control and AdipoVD following cold exposure. (A–E) n = 10 control/ n = 7 AdipoVD. Blue = DAPI. Bars = 100 µm.

Article Snippet: Neurotensin protein levels from tissue lysates were measured by Neurotensin EIA kit (RayBiotech) according to manufacturer’s protocols and normalized to the total protein concentration.

Techniques: Control, Gene Expression

Journal: Frontiers in cell and developmental biology

Article Title: Increased adipose tissue lymphatic vessel density inhibits thermogenesis through elevated neurotensin levels.

doi: 10.3389/fcell.2023.1100788

Figure Lengend Snippet: FIGURE 4 Inhibition of neurotensin activity ameliorate temperature dysfunction in AdipoVD mice. (A) Body temperature of Control and AdipoVD mice during acute under cold exposure at 4°C after neurotensin receptor inhibition. (B) Browning and thermogenesis-associated gene expression in AdipoVD BAT relative to Controls under acute cold exposure after neurotensin receptor inhibition. (C) Immunofluorescence of UCP-1 (green) and lymphatics (red; LYVE-1) from BAT depot of Control and AdipoVD following cold exposure. (D) Browning and thermogenesis-associated gene expression in AdipoVD SQAT relative to Controls following cold exposure and neurotensin receptor inhibition. (E) Immunofluorescence of UCP-1 (green) and lymphatics (red; LYVE-1) from SQAT depot of Control and AdipoVD following cold exposure and neurotensin receptor inhibition. (A–E) n = 10 control/n = 7 AdipoVD. Blue = DAPI. Bars = 100 µm.

Article Snippet: Neurotensin protein levels from tissue lysates were measured by Neurotensin EIA kit (RayBiotech) according to manufacturer’s protocols and normalized to the total protein concentration.

Techniques: Inhibition, Activity Assay, Control, Gene Expression

Journal: Molecular psychiatry

Article Title: Ankyrin-G isoform imbalance and interneuronopathy link epilepsy and bipolar disorder

doi: 10.1038/mp.2016.233

Figure Lengend Snippet: NT3-AnkG localizes Nav channels to the AIS of PV-interneurons. ( a ) ANK3 gene map labeled with BD-associated SNPs (*) and a BD-protective SNP(*). Alternative exons encoding NT1 (exon 1a and 1a′), NT2 (exon 1e) and NT3 (exon 1b) are labeled. ( b ) Brain AnkG protein isoforms generated by alternative first exon use (NT2 and NT3) and alternative splicing of the large exon (190, 270 and 480 kD). ( c–j ) NT2-AnkG and NT3-AnkG have distinct cell type specific expression in the hippocampus. ( c, e ) Low magnification: CA regions appear to contain only NT2-AnkG, while the dentate gyrus (DG) labels for both NT2- and NT3-AnkG. ( d, f ) Higher magnification (boxed regions from a, c): DG cells express NT2-AnkG always and NT3 variably. ( f, i ) High magnification: CA3 shows labeling of PV interneuron (PV; white) AISs by only NT3 (arrows). CA3 pyramidal cells (PC) express only NT2-AnkG. ( h, j ) High magnification (boxed regions from g , i ): CA3 PVs express only NT3 and PCs only NT2 ( k–l ) Heterozygous Ank3 exon 1b deletion proportionally reduces AnkG and Na V channel labeling at AISs of CA1 PV interneurons. ( k ) CA1 images from Ank3-1b KO/+ and WT mice, acquired under identical conditions. ( l ) Fluorescence intensity at PV AISs is significantly reduced for PanAnkG (p=0.0009), NT3-AnkG (p<0.0001) and PanNa V (p=0.0049) in Ank3-1b KO/+ compared WT. Scales: c, e : 100 μm; d, f, h, j : 10 μm; g, i : 50 μm; d, f subpanels: 5μm; h, j subpanels: 2 μm, k : 10 μm.

Article Snippet: Isoform-specific rabbit antibodies against portions of the NT1-AnkG, NT2-AnkG and NT3-AnkG peptides were raised (YenZym, Inc., S. San Francisco, CA) and affinity-purified using methods described previously( ).

Techniques: Labeling, Generated, Alternative Splicing, Expressing, Fluorescence

Journal: Molecular psychiatry

Article Title: Ankyrin-G isoform imbalance and interneuronopathy link epilepsy and bipolar disorder

doi: 10.1038/mp.2016.233

Figure Lengend Snippet: Cell type specific NT2-AnkG and NT3- AnkG expression is conserved in mouse and human cortex. ( a–f ) Mouse anterior cingulate cortex: ( a–b ) Widespread NT3 ( a : green) and NT2 ( b : green) labeling of neurons in area Cg2/24b. ( c, e ) High magnification: exclusive NT3 expression neocortical PV interneurons at AISs. ( d ) PCs show varied NT3 expression: weak NT3 labeling (◀), moderate NT3 labeling (◀◀)and strong NT3 labeling (◀◀◀). ( f ) PCs show varied NT2 expression: lack NT2 labeling (◀), moderate NT2 labeling ◀◀) and strong NT2 labeling (◀◀◀). ( g–l ) NT2 and NT3 expression pattern is conserved in human. ( g, j ) Human PV interneurons (frontal lateral neocortex) exclusively express NT3. Arrowheads in j mark a PV interneuron AIS lacking NT2 staining. ( h–i, k–l ) Pyramidal neurons variably express the two isoforms. Scales: a, b: 100 μm; c–f, g–l : 10 μm.

Article Snippet: Isoform-specific rabbit antibodies against portions of the NT1-AnkG, NT2-AnkG and NT3-AnkG peptides were raised (YenZym, Inc., S. San Francisco, CA) and affinity-purified using methods described previously( ).

Techniques: Expressing, Labeling, Staining

Journal: Molecular psychiatry

Article Title: Ankyrin-G isoform imbalance and interneuronopathy link epilepsy and bipolar disorder

doi: 10.1038/mp.2016.233

Figure Lengend Snippet: PV-interneurons in the basolateral amygdala (bla) express only NT3-AnkG. ( a ) BLA section labeled by the three indicated antibodies and DAPI. PV interneuron AISs (green arrows) appear yellow due to colabeling by NT3-AnkG (green) and all-AnkG (red) antibodies. Both strongly and weakly PV labeled interneurons (yellow boxes) express NT3. ( b–c ) Higher magnification views of two strongly NT3 labeled boxed cells from a exhibit strong ( b ) and light ( c ) PV labeling. ( d ) Two neighboring non-PV neurons: the upper AIS (single arrowhead) lacks detectable NT3 staining; the lower (double arrowheads) shows light NT3 labeling. Upper panel: merge; middle: NT2; lower: NT3. ( e ) AISs of PV interneurons (red arrows) are not labeled by NT2; both strongly and weakly PV labeled interneurons (yellow boxes) lack NT2. ( f–g ) Higher magnification views of two boxed cells from e, strongly ( f ) and lightly ( g ) PV labeled. Neither AIS stains for NT2. h, Two neighboring non-PV neurons: the upper AIS (single arrowhead) has light NT2 staining; the lower (double arrowheads) shows strong NT2 labeling. Upper panel: merge; middle: NT2; lower: NT3. Scales: a, e: 25 μm; b–d, g–h: 5 μm.

Article Snippet: Isoform-specific rabbit antibodies against portions of the NT1-AnkG, NT2-AnkG and NT3-AnkG peptides were raised (YenZym, Inc., S. San Francisco, CA) and affinity-purified using methods described previously( ).

Techniques: Labeling, Staining