Neurotensin receptor down-regulation induced by dexamethasone and forskolin in rat hypothalamic cultures is mediated by endogenous neurotensin.
Neurotensin (NT) has been shown to be involved in neuroendocrine regulation, and the presence of both the peptide and its receptors has been demonstrated in the hypothalamus. In the present study, we show that hypothalamic neurons in primary cultures express the neurotensin receptor (NTR) and we examined a possible regulation of this receptor by glucocorticoids and activators of adenylate cyclase. In the hypothalamic cultures, 125I-NT bound to a single class of binding sites, presenting a selectivity similar to that observed for the high-affinity NTR previously described in the adult rat brain. Radioautographic studies demonstrated that these 125I-NT binding sites were present on 3% of the neurons. A 48-h treatment with forskolin (fsk) decreased 125I-NT binding by 30%. No effect of dexamethasone (dex) alone was found on that parameter. However, a combined treatment with both agents led to a 40% decrease in 125I-NT binding, corresponding to a reduced number of binding sites, and to a 68% decrease in the amount of NTR mRNA. In parallel, the dex plus forsk treatment increased NT release in the incubation medium. Moreover, the decreases in 125I-NT binding and NTR mRNA induced by this treatment were abolished in the presence of an anti-NT antibody or SR 48692, a non-peptidic antagonist of NTR, suggesting that the down-regulation of NTR observed after dex plus fsk treatment was mediated by the release of endogenous NT. Agonist-induced down-regulation of the NTR in this system was confirmed by the application of an exogenous NT analogue, JMV 449. The present findings indicate that, in hypothalamic cultures, dex and fsk indirectly down-regulate NTR expression via the release of endogenous NT.
Scarcériaux
V
V
INSERM U339, Hôpital Saint-Antoine, Paris, France.
Souazé
F
F
Bachelet
C M
CM
Forgez
P
P
Bourdel
E
E
Martinez
J
J
Rostène
W
W
Pélaprat
D
D
D000704:Analysis of Variance; D000818:Animals; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects; D005938:Glucocorticoids / Q000494:pharmacology*; D007031:Hypothalamus / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D007136:Immunoglobulins / Q000276:immunology; D009474:Neurons / Q000187:drug effects; D009496:Neurotensin / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D018028:Receptors, Neurotensin / Q000032:analysis / Q000187:drug effects* / Q000276:immunology
D005938:Glucocorticoids; D007136:Immunoglobulins; D018028:Receptors, Neurotensin; D005576:Colforsin; D009496:Neurotensin; D003907:Dexamethasone