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Science 9 February 2007:
Vol. 315. no. 5813, p. 766
DOI: 10.1126/science.1135047
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Technical Comments

Comment on "Obestatin, a Peptide Encoded by the Ghrelin Gene, Opposes Ghrelin's Effects on Food Intake"

N. Chartrel1*, R. Alvear-Perez2*, J. Leprince1, X. Iturrioz2, A. Reaux-Le Goazigo2, V. Audinot3, P. Chomarat3, F. Coge3, O. Nosjean3, M. Rodriguez3, J. P. Galizzi3, J. A. Boutin3{dagger}, H. Vaudry1{dagger} and C. Llorens-Cortes2{dagger}

1 Institut National de la Santé et de la Recherche Médicale (INSERM), U413, Laboratory of Cellular and Molecular Neuroendocrinology, and European Institute for Peptide Research (IFRMP 23), University of Rouen, 76821 Mont-Saint-Aignan, France.
2 INSERM, U691, and Collège de France, 75005 Paris, France.
3 Institut de Recherches Servier (IdRS), Centre de Recherches de Croissy, 125 Chemin de la Ronde, 78290 Croissy-sur-Seine, France.


Figure 1 Fig. 1. Binding of obestatin on GPR39-transfected CHO cell membranes and effects of obestatin on cAMP production in CHO cells stably or transiently expressing GPR39. (A) Membranes from transfected CHO cells or from pituitary cells were tested for their ability to bind I125-obestatin using the protocol described by Zhang et al. (1): (1) naive CHO cells, (2) flag-tagged GPR39-expressing CHO cells, (3) EGFP-tagged GPR39-expressing CHO cells, and (4) pituitary cell membranes. Experiments were performed in triplicate and reproduced independently twice. Black bars, total binding; open bars, 10–6 M obestatin. (B to D) CHO cells stably expressing either the EGFP-tagged (B) or the Flag-tagged (C) GPR39, or CHO cells transiently transfected with the human untagged GPR39 (D) were incubated for 1 or 16 hours in the absence or presence of 10–6 or 10–5 M obestatin before cAMP production assessment. [View Larger Version of this Image (38K GIF file)]
 

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