Synapses are central to neuronal signaling and prime targets for drug treatments of neurological and mental disorders. Norepinephrine (NE) regulates attention and alertness. The ? adrenergic receptor ( ? AR) is emerging as the prevalent postsynaptic NE effector at glutamatergic synapses, where it interacts with AMPAR, NMDAR and the postsynaptic L-type Ca2+ channel Cav1.2. These complexes also contain Gs, adenylyl cyclases (ACs) and PKA, the downstream effectors of ? AR, for what appears to be highly localized signaling (within 100 nm) by cAMP (e.g., our work in Science 293, 98;Science 293, 2205;EMBO J 29, 482). Such spatial restriction would explain specific regulation of certain targets of the ? AR - Gs - AC - cAMP - PKA cascade and especially of AMPAR, NMDAR and Cav1.2. This project takes advantage of unique features of glutamatergic postsynaptic sites, which are formed by dendritic spines. AMPAR, NMDAR and Cav1.2 are localized at spine heads by a protein meshwork, the postsynaptic density (PSD), which is small (~300 nm) and can be isolated biochemically.
Aim 1 is to test on a molecular level the hypothesis that specific acute or genetic disruption of the ? AR-AMPAR/NMDAR association affects ? AR-induced phosphorylation of these receptors but not of Cav1.2 that is co-localized within the very same PSDs (PSDs will be immunoprecipitated with antibodies against AMPAR, NMDAR or Cav1.2 for subsequent phospho-analysis of all 3 channels). The ? AR- Cav1.2 binding will be disrupted to test the reverse.
Aim 2 will functionally monitor by high resolution Ca2+ imaging ? AR-stimulated Ca2+ influx through NMDAR and Cav1.2 within same spines with the hypothesis that disrupting ? AR - NMDAR binding will only inhibit ? AR-stimulated Ca2+ influx through NMDAR but not Cav1.2 ? AR (and vice versa).
Aim 3 is to test on a systemic level whether ? AR binding to glutamate receptors, to Cav1.2, or both are important for regulation of a form of LTP induced by a tetanus of 5 Hz (endogenous theta rhythm) for 180 s that requires stimulation of the ? AR and Cav1.2 activity. This work will define unexplored fundamental molecular mechanisms of how NE regulates postsynaptic functions. It will thereby create a framework for understanding neurological diseases such as Alzheimer's disease, which is at least in part due to dysregulation of Cav1.2 and NMDAR by ? AR signaling, and stroke induced neuronal damage, which is at least in part due to upregulation of Ca2+ permeable AMPAR, which in turn are targeted to postsynaptic sites by ? AR signaling. NE signaling is also relevant for PTSD and depression. The postsynaptic assembly of specific signaling components that control PKA-mediated phosphorylation of AMPAR, NMDAR and Cav1.2 constitutes a potentially effective and specific target for drugs that disrupt some of these interactions while not affecting others. Finally, this work will address the question of how localized cAMP signaling can be, which might be <100 nm given the small size of postsynaptic sites. Because ? ARs also associate with Cav1.2 in heart, smooth muscle and pancreas, spatially restricted cAMP signaling is of wide interest beyond its role in the brain.

Public Health Relevance

This project is to investigate the role of physical interactions between proteins that mediate signaling by norepinephrine at synapses, the contact points between neurons where they transmit their signals, typically via glutamate. Aberrant functioning of norepinephrine signaling, of glutamate receptors, and of the Ca2+ channel Cav1.2 are implicated in mental and neurological diseases such as posttraumatic stress disorder, autism, depression, and Alzheimer's disease. Defining new molecular aspects of NE signaling will identify important new drug targets for treatment of these diseases.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Research Project (R01)
Project #
Application #
Study Section
Synapses, Cytoskeleton and Trafficking Study Section (SYN)
Program Officer
Stewart, Randall R
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of California Davis
Schools of Medicine
United States
Zip Code
Nieves-CintrĂ³n, Madeline; Syed, Arsalan U; Buonarati, Olivia R et al. (2017) Impaired BKCa channel function in native vascular smooth muscle from humans with type 2 diabetes. Sci Rep 7:14058
Nystoriak, Matthew A; Nieves-CintrĂ³n, Madeline; Patriarchi, Tommaso et al. (2017) Ser1928 phosphorylation by PKA stimulates the L-type Ca2+ channel CaV1.2 and vasoconstriction during acute hyperglycemia and diabetes. Sci Signal 10:
Qian, Hai; Patriarchi, Tommaso; Price, Jennifer L et al. (2017) Phosphorylation of Ser1928 mediates the enhanced activity of the L-type Ca2+ channel Cav1.2 by the ?2-adrenergic receptor in neurons. Sci Signal 10:
Hell, Johannes W; Navedo, Manuel F; VanHook, Annalisa M (2017) Science Signaling Podcast for 24 January 2017: Tissue-specific regulation of L-type calcium channels. Sci Signal 10:
Tseng, Pang-Yen; Henderson, Peter B; Hergarden, Anne C et al. (2017) ?-Actinin Promotes Surface Localization and Current Density of the Ca2+ Channel CaV1.2 by Binding to the IQ Region of the ?1 Subunit. Biochemistry 56:3669-3681
Patriarchi, Tommaso; Qian, Hai; Di Biase, Valentina et al. (2016) Phosphorylation of Cav1.2 on S1928 uncouples the L-type Ca2+ channel from the ?2 adrenergic receptor. EMBO J 35:1330-45
Chenaux, George; Matt, Lucas; Hill, Travis C et al. (2016) Loss of SynDIG1 Reduces Excitatory Synapse Maturation But Not Formation In Vivo. eNeuro 3:
Patriarchi, Tommaso; Amabile, Sonia; Frullanti, Elisa et al. (2016) Imbalance of excitatory/inhibitory synaptic protein expression in iPSC-derived neurons from FOXG1(+/-) patients and in foxg1(+/-) mice. Eur J Hum Genet 24:871-80
Barcomb, Kelsey; Hell, Johannes W; Benke, Tim A et al. (2016) The CaMKII/GluN2B Protein Interaction Maintains Synaptic Strength. J Biol Chem 291:16082-9
Livide, Gabriella; Patriarchi, Tommaso; Amenduni, Mariangela et al. (2015) GluD1 is a common altered player in neuronal differentiation from both MECP2-mutated and CDKL5-mutated iPS cells. Eur J Hum Genet 23:195-201

Showing the most recent 10 out of 16 publications