Dopamine D1-Like Receptor Stimulation Induces CREB, Arc, and BDNF Dynamic Changes in Differentiated SH-SY5Y Cells
Omar B Rivera-Maya1, Christian D Ortiz-Robles1, José R Palacios-Valladares1
1Department of Toxicology, Center for Research and Advanced Studies of the National Polytechnic Institute, Mexico City, Mexico.
Abstract:
The dopamine D1-like receptor is a dopamine (DA) receptor regulating diverse brain functions. Once the dopamine D1-like receptor is activated, it induces activation of the Protein Kinase A (PKA) that phosphorylates the cAMP Response Element-Binding (CREB) transcription factor, which once active elicits the expression of the critical synaptic elements Activity-regulated cytoskeleton-associated (Arc) and the Brain-Derived Neurotrophic Factor (BDNF). The temporality and subcellular localization of proteins impact brain function. However, there is no information about the temporality of CREB activation and Arc and BDNF levels induced through dopamine D1-like receptor activation. In this study, we aimed to assess the specific effect of dopamine D1-like receptor activation on the temporality of CREB-phosphorylation (p-CREBS133) and the spatiotemporal induction of Arc and BDNF. Using SY-SY5Y cells differentiated with Retinoic Acid (RA), the dopamine D1-like receptor activation with a specific agonist transiently increased p-CREBS133 at 30 min of stimulation. It induced two spikes of Arc protein at 15 min and 6 h, forming clusters near the cell membrane. BDNF secretion temporarily increased, reaching a maximum at 6 h, while secretion was lower at 24 h compared to the unstimulated group. Our results provide new insight into the role of dopamine D1-like receptor activation on CREB activation, Arc, and BDNF increase, showing that these effects occur temporally and for Arc in subcellular specific sites. This study highlights the dopaminergic system as a critical regulator of subcellular events relevant to neuron plasticity. Future research should address the study of the implications for brain function and behavior.
Insights
Dopamine D1-like receptor activation influences CREB phosphorylation and the expression of Arc and BDNF proteins over time. This study reveals the temporal dynamics of these key proteins, offering insights into neural plasticity.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- The dopamine D1-like receptor regulates crucial brain functions.
- Activation of this receptor triggers a signaling cascade involving Protein Kinase A (PKA), cAMP Response Element-Binding (CREB), Activity-regulated cytoskeleton-associated (Arc), and Brain-Derived Neurotrophic Factor (BDNF).
- The temporal and spatial aspects of these molecular events are critical for brain function but remain largely uncharacterized following dopamine D1-like receptor activation.
Purpose of the Study:
- To investigate the precise temporality of CREB phosphorylation (p-CREBS133) induced by dopamine D1-like receptor activation.
- To examine the spatiotemporal induction patterns of Arc and BDNF proteins following dopamine D1-like receptor stimulation.
- To elucidate the role of the dopaminergic system in regulating subcellular events relevant to neuronal plasticity.
Main Methods:
- Utilized SY-SY5Y neuroblastoma cells differentiated with Retinoic Acid (RA).
- Administered a specific agonist to activate dopamine D1-like receptors.
- Assessed p-CREBS133 levels at 30 minutes post-stimulation.
- Quantified Arc protein levels and localization at 15 minutes and 6 hours.
- Measured BDNF secretion at 6 hours and 24 hours post-stimulation.
Main Results:
- Dopamine D1-like receptor activation led to a transient increase in p-CREBS133 at 30 minutes.
- Arc protein exhibited two distinct peaks of expression at 15 minutes and 6 hours, with localization near the cell membrane.
- BDNF secretion significantly increased, peaking at 6 hours, and remained elevated but lower at 24 hours compared to unstimulated controls.
Conclusions:
- Dopamine D1-like receptor activation modulates CREB phosphorylation, Arc, and BDNF expression in a time-dependent manner.
- Arc protein induction occurs at specific subcellular locations, suggesting targeted regulation.
- These findings underscore the dopaminergic system's role in controlling temporal and spatial protein dynamics crucial for neuronal plasticity.
