1. Volume 25, Issue 6, Pages 1390-1399.e6 (June 2017)
Mitochondrial Dynamics Mediated by Mitofusin 1 Is Required for POMC Neuron Glucose-Sensing and Insulin Release Control 
Sara Ramírez, Alicia G. Gómez-Valadés, Marc Schneeberger, Luis Varela, Roberta Haddad-Tóvolli, Jordi Altirriba, Eduard Noguera, Anne Drougard, Álvaro Flores-Martínez, Mónica Imbernón, Iñigo Chivite, Macarena Pozo, Andrés Vidal-Itriago, Ainhoa Garcia, Sara Cervantes, Rosa Gasa, Ruben Nogueiras, Pau Gama-Pérez, Pablo M. Garcia-Roves, David A. Cano, Claude Knauf, Joan-Marc Servitja, Tamas L. Horvath, Ramon Gomis, Antonio Zorzano, Marc Claret 
Cell Metabolism 
Volume 25, Issue 6, Pages e6 (June 2017)
DOI: /j.cmet
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2. Cell Metabolism 2017 25, 1390-1399.e6DOI: (10.1016/j.cmet.2017.05.010)
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3. Figure 1
Intact Mitochondrial Dynamics in POMC Neurons Is Required for Glucose Sensing and Fast to Fed Transition
(A–D) POMC neuron electron microscopy images (A) (bar, 500 nm), mitochondrial density (B), coverage (C) (n = 29–31 neurons/5 mice/genotype), and area (D) (>1,335 mitochondria/5 mice/genotype were analyzed).
(E) POMC neuron mitochondrial aspect ratio (AR) under fed and fasting conditions (n = 60–107 neurons/3–4 mice/genotype/condition). Representative skeletonized images of POMC neuron mitochondrial network are shown.
(F) Volcano plot of hypothalamic gene expression in control and POMCMfn1KO mice. Dashed lines represent the threshold for fold-change (±1.4) and false discovery rate (FDR) (≤0.05). Differentially expressed genes during the fast to fed transition are depicted in red. The number of down and upregulated genes are stated in the upper left and right side, respectively. Unchanged genes are represented in gray. n = 3–4 mice/genotype/condition.
(G) Top enriched functional categories and number of genes differentially expressed.
(H and I) Food intake after i.c.v. vehicle (V) (H and I), glucose (G) (H), or 2-DG (I) administration (n = 5–7/genotype/treatment).
(J) Quantification of C-FOS localization in POMC neurons after vehicle (V) or glucose (G) (n = 2–3/genotype).
All studies have been conducted in control and POMCMfn1KO mice between 10 and 14 weeks of age. Data are expressed as mean ± SEM. ∗p < 0.05; ∗∗∗p < See also Figure S1 and Tables S1 and S2.
Cell Metabolism  , e6DOI: ( /j.cmet )
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4. Figure 2
Defective Glucose Metabolism in POMCMfn1KO Mice Is Caused by Abnormal Insulin Release
(A) Glucose tolerance test (n = 6–11/genotype).
(B) Insulin sensitivity test (n = 7/genotype).
(C) GSIS (n = 6/genotype).
(D) Arginine-stimulated insulin release (n = 5/genotype).
(E) Representative immunofluorescence images displaying glucagon (red) and insulin (green) staining in pancreatic sections. α and β cell mass are shown (n = 3/genotype).
(F) β cell size distribution frequency (n = 3/genotype).
(G) Ex vivo islet GSIS (n = 5/genotype). The average of two independent experiments is shown.
(H) Basal and insulin-induced plasma glucagon levels (n = 7–8/genotype).
(I) Glucagon challenge test (n = 4–5/genotype).
(J and K) Fasting glycemia and HOMA-IR values in mice fed with standard (J) or high-fat diet (K) (n = 5–7/genotype/diet).
(L) Pancreatic islet sympathetic (TH) and parasympathetic (VAChT) nerve density (n = 4–5/genotype).
(M) GSIS after vehicle (V) or Idazoxan (IDA) administration (n = 6/genotype/treatment).
(N) GSIS after vehicle (V) or Carbachol (CAR) administration (n = 6/genotype/treatment).
All studies have been conducted in control and POMCMfn1KO mice between 10 and 14 weeks of age or otherwise stated. Data are expressed as mean ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗∗p < NS: non-significant. See also Figures S2 and S3.
Cell Metabolism  , e6DOI: ( /j.cmet )
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5. Figure 3
Defective GSIS in POMCMfn1KO Mice Is Caused by Enhanced Hypothalamic ROS Production
(A) High-resolution mitochondrial respirometry in ARC-enriched samples from fed (n = 9/genotype) or fasted (n = 4–5/genotype) mice.
(B) Fluorometric ROS production in hypothalamic explants (n = 7–8/genotype/condition).
(C) Amperometric ROS production in hypothalamic explants (n = 4/genotype/condition).
(D) Hypothalamic carbonyl protein content (n = 6–8/genotype).
(E) Neuropeptide hypothalamic content (n = 5–6/genotype).
(F) α-MSH immunoreactivity in the PVN and integrated density (n = 2–3/genotype).
(G) Effects of acute i.c.v. honokiol on GSIS (n = 5–7/genotype/treatment). V, vehicle.
(H) Effects of acute i.c.v. N-acetylcisteine (NaC) on GSIS (n = 5–7/genotype/treatment). V, vehicle.
(I) Hypothalamic expression of ER stress markers measured by qPCR (n = 6–7/genotype).
(J) Effects of acute i.c.v. TUDCA on GSIS (n = 5–6/genotype/treatment). V, vehicle.
All studies have been conducted in control and POMCMfn1KO mice between 10 and 16 weeks of age. Data are expressed as mean ± SEM. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p <
Cell Metabolism  , e6DOI: ( /j.cmet )
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6. Figure 4
Deletion of Opa1 in POMC Neurons Does Not Recapitulate POMCMfn1KO Phenotype
(A) Body weight between 4 and 6 weeks of age (n = 13–20/genotype/age).
(B) Aspect ratio of POMC neuron mitochondria (n = 67–108 neurons/3–4 mice/genotype). Representative skeletonized images of POMC neuron mitochondrial network are shown.
(C and D) Food intake after vehicle (V), glucose (G), or 2-DG administration (n = 8–13/genotype/treatment).
(E) Quantification of C-FOS localization in POMC neurons after vehicle (V) or glucose (G) (n = 4/genotype).
(F) GSIS (n = 6–10/genotype).
(G) Fluorometric ROS production in hypothalamic explants (n = 7–10/genotype/condition).
All studies have been conducted in control and POMCOpa1KO mice between 4 and 6 weeks of age. Data are expressed as mean ± SEM. NS: non-significant. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < See also Figure S4.
Cell Metabolism  , e6DOI: ( /j.cmet )
Copyright © 2017 Elsevier Inc. Terms and Conditions