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Prenatal Alcohol Exposure Leads to Enhanced Serine 9 Phosphorylation of Glycogen Synthase Kinase-3β (GSK-3β) in the Hippocampal Dentate Gyrus of Adult Mouse.
Alcoholism, Clinical and Experimental Research 2017 November
BACKGROUND: The goal of this study was to evaluate the expression and serine 9 phosphorylation of glycogen synthase kinase (GSK-3β) within the adult hippocampal dentate gyrus (DG) in a preclinical mouse model of fetal alcohol spectrum disorders. GSK-3β is a multifunctional kinase that modulates many hippocampal processes affected by gestational alcohol, including synaptic plasticity and adult neurogenesis. GSK-3β is a constitutively active kinase that is negatively regulated by phosphorylation at the serine 9 residue.
METHODS: We utilized a well-characterized limited access "drinking-in-the-dark" paradigm of prenatal alcohol exposure (PAE) and measured p(Ser9)GSK-3β and total GSK-3β within adult DG by Western blot analysis. In addition, we evaluated the expression pattern of both p(Ser9)GSK-3β and total GSK-3β within the adult hippocampal dentate of PAE and control mice using high-resolution confocal microscopy.
RESULTS: Our findings demonstrate a marked 2.0-fold elevation of p(Ser9)GSK-3β in PAE mice, concomitant with a more moderate 36% increase in total GSK-3β. This resulted in an approximate 63% increase in the p(Ser9)GSK-3β/GSK-3β ratio. Immunostaining revealed robust GSK-3β expression within Cornu Ammonis (CA) pyramidal neurons, hilar mossy cells, and a subset of GABAergic interneurons, with low levels of expression within hippocampal progenitors and dentate granule cells.
CONCLUSIONS: These findings suggest that PAE may lead to a long-term disruption of GSK-3β signaling within the DG, and implicate mossy cells, GABAergic interneurons, and CA primary neurons as major targets of this dysregulation.
METHODS: We utilized a well-characterized limited access "drinking-in-the-dark" paradigm of prenatal alcohol exposure (PAE) and measured p(Ser9)GSK-3β and total GSK-3β within adult DG by Western blot analysis. In addition, we evaluated the expression pattern of both p(Ser9)GSK-3β and total GSK-3β within the adult hippocampal dentate of PAE and control mice using high-resolution confocal microscopy.
RESULTS: Our findings demonstrate a marked 2.0-fold elevation of p(Ser9)GSK-3β in PAE mice, concomitant with a more moderate 36% increase in total GSK-3β. This resulted in an approximate 63% increase in the p(Ser9)GSK-3β/GSK-3β ratio. Immunostaining revealed robust GSK-3β expression within Cornu Ammonis (CA) pyramidal neurons, hilar mossy cells, and a subset of GABAergic interneurons, with low levels of expression within hippocampal progenitors and dentate granule cells.
CONCLUSIONS: These findings suggest that PAE may lead to a long-term disruption of GSK-3β signaling within the DG, and implicate mossy cells, GABAergic interneurons, and CA primary neurons as major targets of this dysregulation.
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