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Journal of Histochemistry and Cytochemistry
Volume 52 (3): 335-346, 2004
Copyright ©The Histochemical Society, Inc.

NT Agonist Regulates Expression of Nuclear High-affinity Neurotensin Receptors

Mireille Toy–Miou–Leong, Claude-Marie Bachelet, Didier Pélaprat, William Rostène and Patricia Forgez

INSERM U482, Hôpital Saint-Antoine (MT–M–L,PF); Service d'Imagerie Cellulaire, IFR70, Hôpital de la Pitié Salpêtrière (C-MB); EA 3512 Faculté de Médecine Xavier Bichat (DP); and INSERM EMI 0350, Hôpital Saint-Antoine (WR), France

Correspondence to: Dr. P. Forgez, INSERM U482, Hôpital Saint-Antoine, 184 rue du Faubourg Saint-Antoine 75571, Paris Cedex 12, France. E-mail: forgez{at}st-antoine.inserm.fr

Neurotensin (NT) exerts multiple functions in the central nervous system and peripheral tissues. Its actions are mainly mediated by a high-affinity G-protein-coupled receptor, the NT-1 receptor. In this study we demonstrated a nuclear NT binding site in different cellular models. We first noted that a large percentage of NT-1 receptor cell body immunoreactivity was located in the nuclear soma and nuclear envelope of rat substantia nigra, a brain area rich in NT-containing axon terminals. The NT-1 receptor was also visualized in purified nuclei from CHO cells stably transfected with NT-1 receptor coupled to the enhanced green fluorescence protein by immunocytochemistry. We observed that both the nuclear envelope and the nuclear soma were labeled, and the labeling intensity significantly increased after NT agonist treatment. These results suggested that NT-1 receptors, present in both the nuclear soma and the nuclear envelope, can be modulated by the ligand. Lastly, [125I]-NT binding experiments performed on isolated nuclei from a human lung cancer cell line endogenously expressing NT-1 receptor and NT, LNM35, revealed the existence of nuclear Gpp(NHp)-sensitive binding sites. These binding sites markedly decreased when cells were chronically treated with an NT-1 receptor antagonist, SR 48692. Taken together, these data suggest that the agonist regulates the expression of nuclear NT-1 receptors. (J Histochem Cytochem 52:335–345, 2004)

Key Words: neurotensin • NT-1 receptor • G-protein-coupled receptor • nuclear envelope • nuclear soma • agonist treatment


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