title: Nuclear diacylglycerol lipase-α in rat brain cortical neurons: evidence of 2-arachidonoylglycerol production in concert with phospholipase C-β activity.
abstract: In this report, we describe the localization of diacylglycerol lipase-α (DAGLα) in nuclei from adult cortical neurons, as assessed by double-immunofluorescence staining of rat brain cortical sections and purified intact nuclei and by western blot analysis of subnuclear fractions. Double-labeling assays using the anti-DAGLα antibody and NeuN combined with Hoechst staining showed that only nuclei of neuronal origin were DAGLα positive. At high resolution, DAGLα-signal displayed a punctate pattern in nuclear subdomains poor in Hoechst's chromatin and lamin B1 staining. In contrast, SC-35- and NeuN-signals (markers of the nuclear speckles) showed a high overlap with DAGLα within specific subdomains of the nuclear matrix. Among the members of the phospholipase C-β (PLCβ) family, PLCβ1, PLCβ2, and PLCβ4 exhibited the same distribution with respect to chromatin, lamin B1, SC-35, and NeuN as that described for DAGLα. Furthermore, by quantifying the basal levels of 2-arachidonoylglycerol (2-AG) by liquid chromatography and mass spectrometry (LC-MS), and by characterizing the pharmacology of its accumulation, we describe the presence of a mechanism for 2-AG production, and its PLCβ/DAGLα-dependent biosynthesis in isolated nuclei. These results extend our knowledge about subcellular distribution of neuronal DAGLα, providing biochemical grounds to hypothesize a role for 2-AG locally produced within the neuronal nucleus.
authors.affiliation: Departamento de Neurociencias, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Gontzal
authors.initials: G
authors.lastname: García del Caño
authors.affiliation: Departamento de Neurociencias, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Xabier
authors.initials: X
authors.lastname: Aretxabala
authors.affiliation: Departamento de Farmacología, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Imanol
authors.initials: I
authors.lastname: González-Burguera
authors.affiliation: Departamento de Farmacología, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Mario
authors.initials: M
authors.lastname: Montaña
authors.affiliation: Departamento de Farmacología, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Maider
authors.initials: M
authors.lastname: López de Jesús
authors.affiliation: Departamento de Farmacología, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: Barrondo
authors.affiliation: Departamento de Química Analítica, Facultad de Farmacia, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Ramón J
authors.initials: RJ
authors.lastname: Barrio
authors.affiliation: Servicio General de Análisis, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Sampedro
authors.affiliation: Departamento de Química Analítica, Facultad de Farmacia, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: M Arantzazu
authors.initials: MA
authors.lastname: Goicolea
authors.affiliation: Departamento de Farmacología, Universidad del País Vasco/Euskal Herriko Unibertsitatea (UPV/EHU), 01006 Vitoria-Gasteiz (Araba), Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Sallés
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000096:biosynthesis*; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism*; D002853:Chromatography, Liquid; D063388:Endocannabinoids / Q000096:biosynthesis*; D005455:Fluorescent Antibody Technique; D005989:Glycerides / Q000096:biosynthesis*; D008071:Lipoprotein Lipase / Q000378:metabolism*; D008297:Male; D009474:Neurons / Q000378:metabolism*; D054799:Phospholipase C beta / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013003:Somatosensory Cortex / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D005989:Glycerides; C094503:glyceryl 2-arachidonate; D008071:Lipoprotein Lipase; D054799:Phospholipase C beta
keywords: 2-AG production; DAGLα; PLCβ1; PLCβ2; PLCβ4; neuronal nuclei