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Abstract

The brain hypothalamus contains certain secreted molecules that are important in regulating feeding behaviour1,2,3. Here we show that nesfatin, corresponding to NEFA/nucleobindin2 (NUCB2), a secreted protein of unknown function, is expressed in the appetite-control hypothalamic nuclei in rats. Intracerebroventricular (i.c.v.) injection of NUCB2 reduces feeding. Rat cerebrospinal fluid contains nesfatin-1, an amino-terminal fragment derived from NUCB2, and its expression is decreased in the hypothalamic paraventricular nucleus under starved conditions. I.c.v. injection of nesfatin-1 decreases food intake in a dose-dependent manner, whereas injection of an antibody neutralizing nesfatin-1 stimulates appetite. In contrast, i.c.v. injection of other possible fragments processed from NUCB2 does not promote satiety, and conversion of NUCB2 to nesfatin-1 is necessary to induce feeding suppression. Chronic i.c.v. injection of nesfatin-1 reduces body weight, whereas rats gain body weight after chronic i.c.v. injection of antisense morpholino oligonucleotide against the gene encoding NUCB2. Nesfatin-1-induced anorexia occurs in Zucker rats with a leptin receptor mutation, and an anti-nesfatin-1 antibody does not block leptin-induced anorexia. In contrast, central injection of α-melanocyte-stimulating hormone elevates NUCB2 gene expression in the paraventricular nucleus, and satiety by nesfatin-1 is abolished by an antagonist of the melanocortin-3/4 receptor. We identify nesfatin-1 as a satiety molecule that is associated with melanocortin signalling in the hypothalamus.

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Acknowledgements

We thank M. Taguchi, M. Yoshiba and E. Kada for technical assistance, and K. Yoshikawa and M. Kojima for supplying mouse NUCB2 cDNA and developing ELISA, respectively. This work was supported in part by grants-in-aid from the Ministry of Health, Labor and Welfare of Japan (to M. Mori). Author Contributions S.O. and H.S. contributed equally to this work.

Author information

Affiliations

  1. Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, Showa-machi, Maebashi 371-8511, Japan

    • Shinsuke Oh-I
    • , Hiroyuki Shimizu
    • , Tetsurou Satoh
    • , Shuichi Okada
    • , Toshihiro Imaki
    • , Koushi Hashimoto
    • , Takafumi Tsuchiya
    • , Tsuyoshi Monden
    • , Kazuhiko Horiguchi
    • , Masanobu Yamada
    •  & Masatomo Mori
  2. Department of Regulation Biology, Saitama University, Shimo-okubo, Saitama 378-8570, Japan

    • Sachika Adachi
    •  & Kinji Inoue
  3. Pharmaceutical Discovery Research Laboratory, Teijin Pharma Limited, Asahigaoka, Hino 191-8512, Japan

    • Hiroshi Eguchi
    •  & Masanori Yamamoto

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Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

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Correspondence to Masatomo Mori.

Supplementary information

PDF files

  1. 1.

    Supplementary Figure 1

    Northern blot of cell lysates using a 32P-labelled NUCB2 cDNA probe (bp 402– 967).

  2. 2.

    Supplementary Figure 2

    Effects of troglitazone (TGZ) on body weight in rats.

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    Supplementary Figure 3

    Western blot of rat hypothalamic extracts using an affinity-purified NUCB2 Ab-L (1:1,600 dilution).

  4. 4.

    Supplementary Figure 4

    Food intake in rats after icv injection of an anti-NUCB2 Ab-L.

  5. 5.

    Supplementary Figure 5

    Putative processing sites of the rat NUCB2/nesfatin and Western blot using affinity-purified nesfatin Ab24 and Ab301.

  6. 6.

    Supplementary Figure 6

    Elution profiles of rat hypothalamic extracts (a) and cerebrospinal fluid extracts (b) on HPLC.

  7. 7.

    Supplementary Figure 7

    Western blot of fractionated samples of cerebrospinal fluid extracts on HPLC using an affinity-purified nesfatin Ab24.

  8. 8.

    Supplementary Figure 8

    Immunohistochemical analysis of the rat hypothalamic Arc and PVN using a nesfatin Ab24.

  9. 9.

    Supplementary Figure 9

    Changes in the fat weights after continuous icv injection of nesfatin-1.

  10. 10.

    Supplementary Figure 10

    Hypothalamic NUCB2 concentrations (estimated by Western blot) 10 days after chronic icv injection of missense and antisense MON (daily 40 μg).

  11. 11.

    Supplementary Figure 11

    Effects of alpha-MSH on NUCB2 gene expression in the hypothalamic PVN.

  12. 12.

    Supplementary Figure 12

    Effects of nesfatin-1 on expression of POMC, AgRP, NPY and CRH genes in the hypothalamic Arc and PVN.

  13. 13.

    Supplementary Figure 13

    Nesfatin-1-induced cAMP formation and calcium influx in cells expressing mouse MC3R or MC4R

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    Supplementary Notes

    This file contains the Supplementary Methods and Supplementary Figure Legends.

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https://doi.org/10.1038/nature05162

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