Optogenetic activation of intracellular adenosine A 2 A receptor signaling in hippocampus is sufficient to trigger CREB phosphorylation and impair memory

Human and animal studies have converged to suggest that caffeine consumption prevents memory deficits in aging and Alzheimer’s disease through the antagonism of adenosine A2A receptors (A2AR). To test if A2AR activation in hippocampus is actually sufficient to impair memory function and to begin elucidating the intracellular pathways operated by A2AR, we have developed a chimeric rhodopsin-A2AR protein (optoA2AR), which retains the extracellular and transmembrane domains of rhodopsin (conferring light responsiveness and eliminating adenosine binding pockets) fused to the intracellular loop of A2AR to confer specific A2AR signaling. The specificity of the optoA2AR signaling was confirmed by light-induced selective enhancement of cAMP and phospho-MAPK (but not cGMP) levels in HEK293 cells, which was abolished by a point mutation at the C-terminal of A2AR. Supporting its physiological relevance, optoA2AR activation and the A2AR agonist CGS21680 produced similar activation of cAMP and phosphoMAPK signaling in HEK293 cells, of pMAPK in nucleus accumbens, of c-Fos/pCREB in hippocampus and similarly enhanced long-term potentiation in hippocampus. Remarkably, optoA2AR activation triggered a preferential phospho-CREB signaling in hippocampus and impaired spatial memory performance while optoA2AR activation in the nucleus accumbens Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms Conflict of Interest: Authors declare no conflict of interest for the work presented in this manuscript. HHS Public Access Author manuscript Mol Psychiatry. Author manuscript; available in PMC 2016 May 01. Published in final edited form as: Mol Psychiatry. 2015 November ; 20(11): 1339–1349. doi:10.1038/mp.2014.182. A uhor M anscript

[1]  B. Yee,et al.  Regulation of Fear Responses by Striatal and Extrastriatal Adenosine A2A Receptors in Forebrain , 2014, Biological Psychiatry.

[2]  R. Cunha,et al.  Adenosine A2A Receptors in Striatal Glutamatergic Terminals and GABAergic Neurons Oppositely Modulate Psychostimulant Action and DARPP-32 Phosphorylation , 2013, PloS one.

[3]  P. Jenner,et al.  Effects of the adenosine A2A antagonist istradefylline on cognitive performance in rats with a 6-OHDA lesion in prefrontal cortex , 2013, Psychopharmacology.

[4]  B. Fredholm,et al.  Adenosine receptors as drug targets — what are the challenges? , 2013, Nature Reviews Drug Discovery.

[5]  R. Wise,et al.  Synaptic and Behavioral Profile of Multiple Glutamatergic Inputs to the Nucleus Accumbens , 2012, Neuron.

[6]  P. Agostinho,et al.  Caffeine Consumption Prevents Diabetes-Induced Memory Impairment and Synaptotoxicity in the Hippocampus of NONcZNO10/LTJ Mice , 2012, PloS one.

[7]  D. J. Cook,et al.  Treatment of stroke with a PSD-95 inhibitor in the gyrencephalic primate brain , 2012, Nature.

[8]  Arthur Christopoulos,et al.  Signalling bias in new drug discovery: detection, quantification and therapeutic impact , 2012, Nature Reviews Drug Discovery.

[9]  J. Feldon,et al.  Selective inactivation of adenosine A(2A) receptors in striatal neurons enhances working memory and reversal learning. , 2011, Learning & memory.

[10]  S. Mundell,et al.  Adenosine receptor desensitization and trafficking. , 2011, Biochimica et biophysica acta.

[11]  M. Freissmuth,et al.  From cradle to twilight: the carboxyl terminus directs the fate of the A(2A)-adenosine receptor. , 2011, Biochimica et biophysica acta.

[12]  Catherine J. Wei,et al.  Normal and abnormal functions of adenosine receptors in the central nervous system revealed by genetic knockout studies. , 2011, Biochimica et biophysica acta.

[13]  Kenji F. Tanaka,et al.  Functional Connectome of the Striatal Medium Spiny Neuron , 2011, The Journal of Neuroscience.

[14]  K. Fuxe,et al.  Characterization of the A2AR-D2R interface: focus on the role of the C-terminal tail and the transmembrane helices. , 2010, Biochemical and biophysical research communications.

[15]  Y. Chern,et al.  The A2A adenosine receptor rescues neuritogenesis impaired by p53 blockage via KIF2A, a kinesin family member , 2010, Developmental neurobiology.

[16]  A. Barco,et al.  CREB's control of intrinsic and synaptic plasticity: implications for CREB-dependent memory models , 2010, Trends in Neurosciences.

[17]  P. Agostinho,et al.  Chronic caffeine consumption prevents memory disturbance in different animal models of memory decline. , 2010, Journal of Alzheimer's disease : JAD.

[18]  P. Marin,et al.  GPCR interacting proteins (GIPs) in the nervous system: Roles in physiology and pathologies. , 2010, Annual review of pharmacology and toxicology.

[19]  D. Souza,et al.  Caffeine and an adenosine A2A receptor antagonist prevent memory impairment and synaptotoxicity in adult rats triggered by a convulsive episode in early life , 2010, Journal of neurochemistry.

[20]  H. Yin,et al.  Genetic Deletion of A2A Adenosine Receptors in the Striatum Selectively Impairs Habit Formation , 2009, The Journal of Neuroscience.

[21]  Xiu Song,et al.  Preferential enhancement of working memory in mice lacking adenosine A2A receptors , 2009, Brain Research.

[22]  A. Köfalvi,et al.  Modification upon aging of the density of presynaptic modulation systems in the hippocampus , 2009, Neurobiology of Aging.

[23]  E. Kandel,et al.  Chronic enhancement of CREB activity in the hippocampus interferes with the retrieval of spatial information. , 2009, Learning & memory.

[24]  Marc Flajolet,et al.  FGF acts as a co-transmitter through adenosine A2A receptor to regulate synaptic plasticity , 2008, Nature Neuroscience.

[25]  C. Müller,et al.  Adenosine A2A receptor blockade prevents memory dysfunction caused by β-amyloid peptides but not by scopolamine or MK-801 , 2008, Experimental Neurology.

[26]  J. Tsien,et al.  A Critical Role of the Adenosine A2A Receptor in Extrastriatal Neurons in Modulating Psychomotor Activity as Revealed by Opposite Phenotypes of Striatum and Forebrain A2A Receptor Knock-Outs , 2008, The Journal of Neuroscience.

[27]  F. Pedata,et al.  Adenosine A2A receptors and brain injury: Broad spectrum of neuroprotection, multifaceted actions and “fine tuning” modulation , 2007, Progress in Neurobiology.

[28]  B. Fredholm,et al.  Aspects of the general biology of adenosine A2A signaling , 2007, Progress in Neurobiology.

[29]  A. Nissinen,et al.  Coffee consumption is inversely associated with cognitive decline in elderly European men: the FINE Study , 2007, European Journal of Clinical Nutrition.

[30]  D. Souza,et al.  Caffeine and adenosine A2a receptor antagonists prevent β-amyloid (25–35)-induced cognitive deficits in mice , 2007, Experimental Neurology.

[31]  L. Zacharia,et al.  Caffeine protects Alzheimer’s mice against cognitive impairment and reduces brain β-amyloid production , 2006, Neuroscience.

[32]  Y. Chern,et al.  Rescue of p53 Blockage by the A2A Adenosine Receptor via a Novel Interacting Protein, Translin-Associated Protein X , 2006, Molecular Pharmacology.

[33]  F. Ciruela,et al.  Presynaptic Control of Striatal Glutamatergic Neurotransmission by Adenosine A1–A2A Receptor Heteromers , 2006, The Journal of Neuroscience.

[34]  Q. Wan,et al.  Disruption of PTEN coupling with 5-HT2C receptors suppresses behavioral responses induced by drugs of abuse , 2006, Nature Medicine.

[35]  K. Deisseroth,et al.  Millisecond-timescale, genetically targeted optical control of neural activity , 2005, Nature Neuroscience.

[36]  R. Prediger,et al.  Caffeine reverses age-related deficits in olfactory discrimination and social recognition memory in rats Involvement of adenosine A1 and A2A receptors , 2005, Neurobiology of Aging.

[37]  F. Lee,et al.  Transactivation of Trk Neurotrophin Receptors by G-Protein-Coupled Receptor Ligands Occurs on Intracellular Membranes , 2004, The Journal of Neuroscience.

[38]  J. Jolles,et al.  The effects of habitual caffeine use on cognitive change: a longitudinal perspective , 2003, Pharmacology Biochemistry and Behavior.

[39]  Yitao Liu,et al.  Treatment of Ischemic Brain Damage by Perturbing NMDA Receptor- PSD-95 Protein Interactions , 2002, Science.

[40]  R. Hébert,et al.  Risk factors for Alzheimer's disease: a prospective analysis from the Canadian Study of Health and Aging. , 2002, American journal of epidemiology.

[41]  R. Cunha,et al.  Parallel modification of adenosine extracellular metabolism and modulatory action in the hippocampus of aged rats , 2001, Journal of neurochemistry.

[42]  J. Jolles,et al.  Habitual caffeine consumption and its relation to memory, attention, planning capacity and psychomotor performance across multiple age groups , 2000, Human psychopharmacology.

[43]  P. Svenningsson,et al.  Distribution, biochemistry and function of striatal adenosine A2A receptors , 1999, Progress in Neurobiology.

[44]  W. Hauber,et al.  Motor depressant effects mediated by dopamine D2 and adenosine A2A receptors in the nucleus accumbens and the caudate-putamen. , 1997, European journal of pharmacology.

[45]  H. Simon,et al.  Extension of a New Two-Trial Memory Task in the Rat: Influence of Environmental Context on Recognition Processes , 1997, Neurobiology of Learning and Memory.

[46]  C. Gerfen,et al.  D1 and D2 dopamine receptor-regulated gene expression of striatonigral and striatopallidal neurons. , 1990, Science.

[47]  R. Cunha,et al.  Caffeine consumption prevents memory impairment, neuronal damage, and adenosine A2A receptors upregulation in the hippocampus of a rat model of sporadic dementia. , 2013, Journal of Alzheimer's disease : JAD.

[48]  L. White,et al.  Coffee intake in midlife and risk of dementia and its neuropathologic correlates. , 2011, Journal of Alzheimer's disease : JAD.

[49]  H. Soininen,et al.  Midlife coffee and tea drinking and the risk of late-life dementia: a population-based CAIDE study. , 2009, Journal of Alzheimer's disease : JAD.

[50]  J. Zezula,et al.  Heterotrimeric G Protein-independent Signaling of a G Protein-coupled Receptor DIRECT BINDING OF ARNO/CYTOHESIN-2 TO THE CARBOXYL TERMINUS OF THE A2A ADENOSINE RECEPTOR IS NECESSARY FOR SUSTAINED ACTIVATION OF THE ERK/MAP KINASE PATHWAY* , 2005 .

[51]  H. Normile,et al.  ROLE OF ADENOSINE A2a RECEPTORS IN THE NUCLEUS ACCUMBENS. , 1994, Progress in neuro-psychopharmacology & biological psychiatry.