NPY-Y1R signalling plays a prominent role in the regulation of several behavioural and physiological functions including feeding behaviour and energy balance, sexual hormone secretion, stress response, emotional behaviour, neuronal excitability and ethanol drinking. Y1R activation has been shown to mobilize intracellular Ca2+ and to inhibit cAMP synthesis in many tissues. Intracellular increase in these second messengers leads to the stimulation of Ca2+CaM kinase (CaMK) and of protein kinase A, respectively that, in turn, activate transcription factors resulting in stimulation or inhibition of target genes. Previous studies demonstrated that the stimulation of Y1R increases cAMP responsive element binding protein (CREB) phosphorylation and induces the expression of cAMP responsive element (CRE) containing genes, including the Y1R gene, via mobilization of Ca2+ and activation of CaMKs. We now investigated the regional distribution and co-localization of Y1R and alpha-CaMKII kinase by using CRE indicator mice. Mice were obtained by breeding a transgenic mouse line expressing a alpha-CaMKII promoter-driven tTA and the Cre reporter line RosaR26, harbouring a LacZ gene silenced with a floxed transcriptional terminator sequence. In these mice, the region specific Cre-mediated removal of the transcriptional terminator sequence activates LacZ gene expression in alpha-CaMKII expressing cells. Immunoreactive staining of Y1R positive neurons and histochemical staining of beta-galactosidase revealed that Y1R and alpha-CaMKII are co-localized in the same areas of the forebrain, with markedly intense labelling for both genes in the cerebral cortex, CA1, CA3, and DG subfields of the hippocampus, striatum and amygdaloid complex. Double labelling for Y1R and LacZ on the same brain slice showed that in all regions Y1R positive neurons co-express X-gal staining. These data provide further evidence that Y1R-activated intracellular responses may involve regulation alpha-CaMKII activity.
Comparative analysis of brain distribution of NPY-Y1 receptor and alpha-CamKII kinase in a Cre indicator mouse
OBERTO, Alessandra;BERTOCCHI, Ilaria;EVA, Carola Eugenia
2006-01-01
Abstract
NPY-Y1R signalling plays a prominent role in the regulation of several behavioural and physiological functions including feeding behaviour and energy balance, sexual hormone secretion, stress response, emotional behaviour, neuronal excitability and ethanol drinking. Y1R activation has been shown to mobilize intracellular Ca2+ and to inhibit cAMP synthesis in many tissues. Intracellular increase in these second messengers leads to the stimulation of Ca2+CaM kinase (CaMK) and of protein kinase A, respectively that, in turn, activate transcription factors resulting in stimulation or inhibition of target genes. Previous studies demonstrated that the stimulation of Y1R increases cAMP responsive element binding protein (CREB) phosphorylation and induces the expression of cAMP responsive element (CRE) containing genes, including the Y1R gene, via mobilization of Ca2+ and activation of CaMKs. We now investigated the regional distribution and co-localization of Y1R and alpha-CaMKII kinase by using CRE indicator mice. Mice were obtained by breeding a transgenic mouse line expressing a alpha-CaMKII promoter-driven tTA and the Cre reporter line RosaR26, harbouring a LacZ gene silenced with a floxed transcriptional terminator sequence. In these mice, the region specific Cre-mediated removal of the transcriptional terminator sequence activates LacZ gene expression in alpha-CaMKII expressing cells. Immunoreactive staining of Y1R positive neurons and histochemical staining of beta-galactosidase revealed that Y1R and alpha-CaMKII are co-localized in the same areas of the forebrain, with markedly intense labelling for both genes in the cerebral cortex, CA1, CA3, and DG subfields of the hippocampus, striatum and amygdaloid complex. Double labelling for Y1R and LacZ on the same brain slice showed that in all regions Y1R positive neurons co-express X-gal staining. These data provide further evidence that Y1R-activated intracellular responses may involve regulation alpha-CaMKII activity.
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