custom fabricated ferrule capped optical fiber  (Thorlabs)

Journal: Cell reports

Article Title: Opioidergic signaling contributes to food-mediated suppression of AgRP neurons

doi: 10.1016/j.celrep.2023.113630

Figure Lengend Snippet: (A) Cartoon depiction of the fluorescent sensor based on the δ-opioid receptor (DOR). (B) Schematic showing injection of the DeltaLight sensor expressing virus into the arcuate nucleus and recording by fiber photometry. (C) Photomicrograph showing ferrule placement (ferrule tip [FT]) over the mediobasal hypothalamus (top) and DeltaLight sensor expression in hypothalamic neurons (bottom right) and a map of FT locations in each mouse (bottom left). Scale bars: 200 μm (top), 30 μm (bottom right). (D and E) Change in DeltaLight sensor activity in response to non-edible object and food presentation to overnight-fasted mice (D) and summarized mean sensor activity in 5-min time bins (E) (n = 6 mice, object vs. chow, paired t test, *p < 0.038). (F and G) Change in DeltaLight and mutant sensor (Ctrl) activity in response to high fat, high sugar (HFHS) presentation in ad libitum -fed mice (F) and summarized mean sensor activity in 5-min bins (G) (n = 4 mice each, *p < 0.037, **p = 0.006, Ctrl vs. DeltaLight, unpaired t test). All data are shown as mean ± SEM.

Article Snippet: For in vivo fiber photometry recording, ferrule capped metal optical fiber (200 μm core diameter, NA = 0.48, Thorlabs) was implanted above the ARC using the same coordinates, except for the dorsal surface, which was ~100–200 mm above the viral injection.

Techniques: Injection, Expressing, Virus, Activity Assay, Mutagenesis

Journal: Cell reports

Article Title: Opioidergic signaling contributes to food-mediated suppression of AgRP neurons

doi: 10.1016/j.celrep.2023.113630

Figure Lengend Snippet: (A) Schematic showing injection of the FLEX-GCaMP7s-expressing virus and ferrule placement over the ARC for fiber photometry recording and micrograph image showing injection and ferrule locations. Scale bar: 150 μm. (B and C) Change in average AgRP neuron activity (B, top) and activity heatmap for individual mice (B, bottom) in response to i.p. injection (vertical dashed line) of saline or DAMGO (1 mg/kg) and summarized mean of AgRP neuron activity in 5-min time bins (C, n = 10 mice, *p < 0.043, a p<0.0084, b p<0.00011, saline vs. DAMGO, paired t test). (D) Schematic showing loose seal recordings from GFP-labeled NPY neurons in the ARC. (E and F) Representative loose seal traces (E) and summary of mean frequency of the recorded neurons (F) before (baseline) or after DAMGO (2 μM) bath application (n = 37–39 neurons, respectively/4 mice each, p < 0.0001, unpaired t test). (G and H) Representative whole-cell current-clamp recording from ARC NPY neurons (G) and resting membrane potential values (H) showing robust hyperpolarization by DAMGO in the presence of synaptic blockers (n = 15 neurons/3 mice, **p < 0.01, ***p < 0.001, paired t test). (I and J) Whole-cell current-clamp recordings from ARC NPY neurons using internal pipette solution with GDPβS instead of GTP with synaptic blockers (n = 11 neurons/3 mice). All data are shown as mean ± SEM.

Article Snippet: For in vivo fiber photometry recording, ferrule capped metal optical fiber (200 μm core diameter, NA = 0.48, Thorlabs) was implanted above the ARC using the same coordinates, except for the dorsal surface, which was ~100–200 mm above the viral injection.

Techniques: Injection, Expressing, Virus, Activity Assay, Saline, Labeling, Membrane, Transferring

Journal: Cell reports

Article Title: Opioidergic signaling contributes to food-mediated suppression of AgRP neurons

doi: 10.1016/j.celrep.2023.113630

Figure Lengend Snippet: (A and B) Schematic showing recording from GFP-positive cells of Npy-gfp mice, representative loose seal recording (A) and summary bar graph of firing rates (B) showing the effect of naloxone (NalX; 2 μM) treatment on ARC NPY neurons in sated mice (n = 31–34 neurons/4 mice, p < 0.0001, unpaired t test). (C and D) In vivo fiber photometry recording from AgRP neurons in fasted mice injected with saline, NalX (4 mg/kg), or CTAP (1.5 mg/kg) 15 min before chow presentation (vertical dashed line, C) and summary graph showing average change in activity in 5-min time bins (D) (n = 10 mice, *p < 0.05, **p < 0.0097, ***p < 0.00095; red asterisks, NalX vs. saline; blue asterisks, CTAP vs. saline; paired t test). (E and F) In vivo fiber photometry recording from AgRP neurons in ad-libitum- fed mice injected with saline (from ), satiety cocktail (3 μg/kg CCK + 10 μg/kg amylin + 10 μg/kg PYY) or satiety cocktail with NalX (vertical dashed line, E), and summary graph showing average change in activity in 5-min time bins (F) (n = 9 mice, satiety cocktail vs. satiety cocktail with NalX, paired t test, *p < 0.04, a p = 0.0084). (G and H) In vivo fiber photometry recording from AgRP neurons in ad libitum -fed mice injected with saline, EtOH (15%), or EtOH with NalX (vertical dashed line, G) and summary graph showing average change in activity in 5-min time bins (H) (n = 8 mice; red asterisks, saline vs. EtOH; blue asterisks, saline vs. EtOH with NalX; paired t test, **p < 0.0095, *p < 0.019). (I and J) DAMGO-induced (I) and morphine-induced (J) suppression of dark-onset feeding (*p = 0.033, ***p < 0.0001, paired t test). All data are shown as mean ± SEM.

Article Snippet: For in vivo fiber photometry recording, ferrule capped metal optical fiber (200 μm core diameter, NA = 0.48, Thorlabs) was implanted above the ARC using the same coordinates, except for the dorsal surface, which was ~100–200 mm above the viral injection.

Techniques: In Vivo, Injection, Saline, Activity Assay

Journal: Cell reports

Article Title: Opioidergic signaling contributes to food-mediated suppression of AgRP neurons

doi: 10.1016/j.celrep.2023.113630

Figure Lengend Snippet: (A) Breeding strategy to generate mice with AgRP neuron-specific ablation of MOR. (B and C) Schematic showing recording from GFP-positive cells in cre-dependent GFP-expressing virus-injected Agrp-ires-cre mice and representative loose seal traces showing effect of DAMGO perfusion in WT and MOR-deficient (AgRP-MKO) AgRP neurons’ firing rates (B) and summary bar graph showing quantification (C). WT, 22–26 neurons; AgRP-MKO, 16 neurons each for BL and DAMGO recordings, respectively. BL vs. DAMGO, unpaired t test, ***p < 0.001; ns, not significant). (D) In vivo fiber photometry recording from AgRP neurons in fasted WT and AgRP-MKO mice during object or chow presentation (vertical dashed line, left) and summary graph showing average change in activity in 5-min time bins (right) (n = 9 WT, 10 AgRP-MKO mice; all time points: not significant for WT vs. AgRP-MKO comparisons; unpaired t test). (E) In vivo fiber photometry recording from AgRP neurons in ad libitum -fed WT and AgRP-MKO mice injected with saline or satiety cocktail (vertical dashed line, left) and summary graph showing average change in activity in 5-min time bins (right) (n = 10 mice each, WT vs. AgRP-MKO, unpaired t test, *p = 0.047). (F) In vivo fiber photometry recording from AgRP neurons in ad libitum -fed WT and AgRP-MKO mice during HFHS presentation (vertical dashed line, left) and summary graph showing average change in activity in 5-min time bins (right) (n = 11 WT, 9 AgRP-MKO mice; t, trend; p < 0.07, unpaired t test). (G) In vivo fiber photometry recording from AgRP neurons in ad libitum -fed WT and AgRP-MKO mice injected with saline or morphine (10 mg/kg, vertical dashed line, left) and summary graph showing average change in activity in 5-min time bins (right) (n = 11 WT, 9 AgRP MKO mice; WT morphine vs. AgRP-MKO morphine, unpaired t test, black *p < 0.046; blue marks, WT; red marks, AgRP-MKO; saline vs. morphine paired t tests, *p < 0.043, ap<0.0091, bp<0.001). (H) In vivo fiber photometry recording from AgRP neurons in fasted AgRP-MKO mice during chow presentation (vertical dashed line, left) after i.p. injection of saline and CTAP and summary graph showing average change in activity in 5-min time bins (right) (n = 3 mice, all time points: not significant, paired t test). (I) In vivo fiber photometry recording from AgRP neurons in ad libitum -fed AgRP MKO mice during i.p. injection of satiety cocktail with and without NalX (vertical dashed line, left) and summary graph showing average change in activity in 5-min time bins (right) (n = 3 mice, all time points: not significant, paired t test). All data are shown as mean ± SEM.

Article Snippet: For in vivo fiber photometry recording, ferrule capped metal optical fiber (200 μm core diameter, NA = 0.48, Thorlabs) was implanted above the ARC using the same coordinates, except for the dorsal surface, which was ~100–200 mm above the viral injection.

Techniques: Expressing, Virus, Injection, In Vivo, Activity Assay, Saline