Expression data from NPY Y1R-deficient osteoblastic cells
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE114469 | GEO | 2018/05/16
REPOSITORIES: GEO
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Project description:Neuropeptide Y (NPY) is a peptide widely distributed throughout the body that is involved in various physiological processes, including the regulation of feeding behavior and energy homeostasis and, as recently reported, the regulation of bone homeostasis. The Y1 receptors have been identified as major contributors to this latter biological effect. 5 - Carbamimidamido- 2 - (2,2 - diphenylacetamido) – N - [(4- hydroxyphenyl)methyl] pentanamide (BIBP 3226) is a selective NPY Y1 receptor antagonist with recognized potential as therapeutic agent in bone regeneration and its quantification in cellular internalization assays was aimed in the present work. Using high-performance liquid chromatography coupled to triple quadrupole-tandem mass spectrometry, BIBP 3226 determination was attained with a reversed phase column, isocratic elution, detection in positive ionization mode and data acquisition in selected reaction monitoring mode. Calibration curves were linear for concentrations ranging from 0.25 to 30 ng mL-1 with LOD and LOQ values as low as 0.1 and 0.3 pg in cell extracts and 16 and 48 pg in supernatant culture media, respectively. BIBP 3226 was successfully quantified in cell extracts and supernatants obtained from internalization assays using bone marrow cells (Wt C57/Bl6 mice) and also breast cancer cells (MCF7), which overexpress the NPY Y1 receptor. Therefore, the determined amount of BIBP 3226 was related to the levels of NPY Y1 receptor, demonstrating the method suitability to measure NPY Y1 receptor expression indirectly as an alternative to antibody-based fluorescence microscopy.
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Project description:Expression data from NPY Y1R-deficient osteoblastic cells
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Project description:Neuropeptide Y (NPY) is an endogenous modulator of neuronal activity by regulating GABA and glutamate release. Previously, we found that estradiol modulates NPY expression in the hippocampal dentate gyrus. Here we investigated which estrogen receptor type activation is required for the NPY expression. Further, we determined effects of estrogen receptor activation on NPY release. Finally, we determined the contribution of estrogen-mediated remodeling of the GABAergic and glutamatergic network in relation to changes in coupling with NPY in ovariectomized rats. We found that activation of either estrogen receptor type increases NPY expression as well as NPY release in the dentate gyrus. We also found that compared to OVX rats, estrogen replacement increases the likeness of synergistic/antagonistic coupling between the NPY and GABAergic synapse genes while the glutamatergic synapse genes are less likely coupled with NPY. The data together suggest that estrogen plays a critical role in regulation of activity of the NPY system and its coupling to GABAergic and glutamatergic synapses in female rat dentate gyrus. Two-conditions (E = beta-estradiol replacement vs O = oil) experiment. Biological replicates: 4E, 4O.
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Project description:Neuropeptide Y (NPY) is an endogenous modulator of neuronal activity by regulating GABA and glutamate release. Previously, we found that estradiol modulates NPY expression in the hippocampal dentate gyrus. Here we investigated which estrogen receptor type activation is required for the NPY expression. Further, we determined effects of estrogen receptor activation on NPY release. Finally, we determined the contribution of estrogen-mediated remodeling of the GABAergic and glutamatergic network in relation to changes in coupling with NPY in ovariectomized rats. We found that activation of either estrogen receptor type increases NPY expression as well as NPY release in the dentate gyrus. We also found that compared to OVX rats, estrogen replacement increases the likeness of synergistic/antagonistic coupling between the NPY and GABAergic synapse genes while the glutamatergic synapse genes are less likely coupled with NPY. The data together suggest that estrogen plays a critical role in regulation of activity of the NPY system and its coupling to GABAergic and glutamatergic synapses in female rat dentate gyrus.
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