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Preferential activation by galanin 1-15 fragment of the GalR1 protomer of a GalR1-GalR2 heteroreceptor complex

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10.1016/j.bbrc.2014.08.061
 
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hdl:2117/26506

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Borroto Escuela, Dasiel Óscar
Narvaez, Manuel
Di Palma, Michael
Calvo, Feliciano
Rodríguez, David
Millón, Carmelo
Carlsson, Jens
Agnati, Luigi F.
Garriga Solé, PereMés informacióMés informacióMés informació
Díaz Cabiale, Zaida
Fuxe, Kjell
Document typeArticle
Defense date2014-09-26
Rights accessRestricted access - publisher's policy
All rights reserved. This work is protected by the corresponding intellectual and industrial property rights. Without prejudice to any existing legal exemptions, reproduction, distribution, public communication or transformation of this work are prohibited without permission of the copyright holder
Abstract
The three cloned galanin receptors show a higher affinity for galanin than for galanin N-terminal fragments. Galanin fragment (1-15) binding sites were discovered in the rat Central Nervous System, especially in dorsal hippocampus, indicating a relevant role of galanin fragments in central galanin communication. The hypothesis was introduced that these N-terminal galanin fragment preferring sites are formed through the formation of GalR1-GalR2 heteromers which may play a significant role in mediating galanin fragment (1-15) signaling. In HEK293T cells evidence for the existence of GalR1-GalR2 heteroreceptor complexes were obtained with proximity ligation and BRET2 assays. PLA positive blobs representing GalR1-GalR2 heteroreceptor complexes were also observed in the raphe-hippocampal system. In CRE luciferase reporter gene assays, galanin (1-15) was more potent than galanin (1-29) in inhibiting the forskolin-induced increase of luciferase activity in GalR1-GalR2 transfected cells. The inhibition of CREB by 50 nM of galanin (1-15) and of galanin (1-29) was fully counteracted by the non-selective galanin antagonist M35 and the selective GalR2 antagonist M871. These results suggested that the orthosteric agonist binding site of GalR1 protomer may have an increased affinity for the galanin (115) vs galanin (1-29) which can lead to its demonstrated increase in potency to inhibit CREB vs galanin (1-29). In contrast, in NFAT reporter gene assays galanin (1-29) shows a higher efficacy than galanin (115) in increasing Gq/11 mediated signaling over the GalR2 of these heteroreceptor complexes. This disbalance in the signaling of the GalR1-GalR2 heteroreceptor complexes induced by galanin (1-15) may contribute to depression-like actions since GalR1 agonists produce such effects. (C) 2014 Elsevier Inc. All rights reserved.
CitationBorroto, D. [et al.]. Preferential activation by galanin 1-15 fragment of the GalR1 protomer of a GalR1-GalR2 heteroreceptor complex. "Biochemical and biophysical research communications", 26 Setembre 2014, vol. 452, núm. 3, p. 347-353. 
URIhttp://hdl.handle.net/2117/26506
DOI10.1016/j.bbrc.2014.08.061
ISSN0006-291X
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  • GBMI - Grup de Biotecnologia Molecular i Industrial - Articles de revista [170]
  • Departament d'Enginyeria Química - Articles de revista [1.989]
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