A growing body of data shows that the AMPA subtype of glutamate receptors play a major role in regulating short and long term forms of synaptic plasticity. Furthermore, it is now clear that regulation of plasticity occurs predominantly by regulating the trafficking of AMPA receptor subunits, and their insertion and removal from the synapse. Notably, this trafficking is primarily dependent upon AMPA receptor subunit phosphorylation by 3 major signaling pathways known to be targets for mood stabilizers: the PKC, PKA and MAPK cascades. In view of the growing body of data suggesting that severe mood disorders may be associated with impairments of cellular plasticity, we undertook the present series of studies to determine if two clinically effective, but structurally highly dissimilar antimanic agents, lithium and VPA, regulate synaptic expression of AMPA receptor subunit GluR1. Administration of chronic lithium or valproate (at therapeutically relevant concentrations) reduced rat hippocampal synaptosomal levels of GluR1 after by 40% and 20%, respectively. In cultured hippocampal neurons, both lithium and VPA significantly down regulated the surface expression of GluR1 40% in a dose and time-dependent manner. Surface staining with an anti-N terminal GluR1 antibody confirmed the result. Double-immunostaining of GluR1 and synaptotagmin showed that chronic treatment attenuated the numbers of GluR1 positive synapses of lithium and valproate-treated neurons. However, total protein levels of GluR1, and synaptotagmin remained unchanged after lithium and valproate treatment in vitro and in vivo. Lithium and valproate treatment also attenuated the phosphorylation of a specific PKA site (GluRp845) by 52 and 33% respectively. Sp-cAMP treatment reversed the attenuation of phosphorylation by lithium and valproate and also brought GluR1s back to the surface, suggesting that phosphorylation of GluRp845 is involved in the mechanism of GluR1 surface attenuation. In striking contrast, drugs, such as imipramine, which induce mania, increase the synaptic expression of GluR1 in vivo in hippocampus. ? In order to develop a new potential drug which mimics the effect of mood stabilizers on GluR1 phosphorylation, TAT-peptides (TAT-p845 and TAT-SRC) were designed and synthesized. Tat peptide (YGRKKRRQRRR) is a leading peptide, which enables the functional peptide to pass through the blood brain barrier and cell membrane, allowing it to get into cytosol or synapses of the neurons. A previous study has successfully utilized peptide injection into animals to disrupt the interaction of PSD-95 with NMDA receptors in the brain and to provide a neuroprotective effect on a stroke animal model. TAT-p845 was able to inhibit the phosphorylation of AMPA receptors at its PKA site and down-regulate the surface expression of GluR1 in cultured hippocampal neurons, which is the same effect produced by lithium and valproate. Moreover, this TAT-p845 was able to pass the blood brain barrier and inhibit the phosphorylation of GluR1 in the hippocampus in vivo, which again demonstrated its ability to induce the same effects as lithium and valproate. In addition, reduction of GluR1 phosphorylation at its PKA site by Tat-p845, was sufficient to attenuate synaptic GluR1/2 in hippocampal neurons in vivo. Intra-hippocampal infusion of AMPA-specific inhibitor GYKI54226, GluR1-specific TAT-p845 peptide and GluR1-PDZ-specific TAT-TGL peptide were able to attenuate amphetamine-induced hyperactivity and/or amphetamine-induced conditioned-place preference in the mania animal model. These studies provide novel mechanisms for anti-manic effect through attenuationof AMPA receptor activity and avenues for new drug development for mood disorders. TAT-p845, which attenuates AMPA receptor levels at synapses, may offer exciting possibilities as a new class of medicine with the potential for treatment of: bipolar disorder. ? ? Chronic treatment with the antimanic agents, lithium and valproate resulted in reduced synaptic expression of the AMPA receptor subunit GluR1 in the hippocampus while treatment with an antidepressant (imipramine) enhanced the synaptic expression of GluR1. The anticonvulsants, lamotrigine and riluzole have been demonstrated to have efficacy in the depressive phase of bipolar disorder. We therefore sought to determine the role of these anticonvulsants, compared to that of the predominantly antimanic anticonvulsant valproate, on AMPA receptor localization. We found that the agents with a predominantly antidepressant profile, namely lamotrigine and riluzole, significantly enhanced the surface expression of GluR1 and GluR2 in a time- and dose-dependent-manner in cultured hippocampal neurons. By contrast, the predominantly antimanic agent, valproate, significantly reduced surface expression of GluR1 and GluR2. Concomitant with the GluR1 and GluR2 changes, the peak value of depolarized membrane potential evoked by AMPA was significantly higher in lamotrigine and riluzole treated neurons, supporting the surface receptor changes. Phosphorylation of GluR1 at the PKA site (S845) was enhanced in both lamotrigine- and riluzole- treated hippocampal neurons, but reduced in valproate treated neurons. In addition, lamotrigine and riluzole, as well as the traditional antidepressant imipramine, also increased GluR1 phosphorylation at GluR1 (S845) in the hippocampus after chronic in vivo treatment. Our findings suggest that regulation of GluR1/2 surface levels and function may be responsible for the different clinical profile of anticonvulsants (antimanic or antidepressant), and may suggest avenues for the development of novel therapeutics for these illnesses.In addition, we found that glutamate receptors are targets of Protein Kinase C (PKC), which plays an important role in the pathophysiology and treatment of mania. Considerable biochemical evidence suggests that the protein kinase C (PKC) signaling cascade may be a convergent point for the actions of anti-manic agents, and that excessive PKC activation can disrupt prefrontal cortical regulation of thinking and behavior. Currently, however, brain protein targets of PKCs anti-manic effects remains unclear. Here we showed that PKC activity was enhanced in the prefrontal cortex brain region of animals treated with the psychostimulant amphetamine and the antidepressant imipramine. Phosphorylation of myristoylated alanine-rich C kinase substrate (MARCKS), a marker of PKC activity, was increased in the prefrontal cortex of psychstimulant amphetamine treated animals, as well as in sleep-deprived animals (another animal model of mania), but decreased in lithium-treated animals. Antidepressant imipramine, which shows promanic property on bipolar patients, also enhanced pMARCKS in prefrontal cortex in vivo. We further explored the functional targets of PKC in mania-associated behaviors. Neurogranin is a brain-specific, postsynaptically located PKC substrate. PKC phosphorylation of neurogranin was robustly increased by pro-manic manipulations and decreased by anti-manic agent. PKC phosphorylation of the NMDA receptor site NR1S896 and the AMPA receptor site GluR1T840 was also enhanced in the prefrontal cortex of animals treated with antidepressant imipramine, as well as behaviorally sleep-deprived, in striking contrast to the reduced activity seen in lithium-treated animals. These results suggest that PKC may play an important role in regulating NMDA and AMPA receptor functions. The biochemical profile of the PKC pathway thus encompasses both pro- and anti-manic effects on behavior. These results suggest that PKC modulators or their intracellular targets may ultimately represent novel avenues for the development of new therapeutics for mood disorders.
| Catapano, Lisa A; Manji, Husseini K (2008) Kinases as drug targets in the treatment of bipolar disorder. Drug Discov Today 13:295-302 |
| Catapano, Lisa A; Manji, Husseini K (2007) G protein-coupled receptors in major psychiatric disorders. Biochim Biophys Acta 1768:976-93 |
| Du, Jing; Suzuki, Katsuji; Wei, Yanling et al. (2007) The anticonvulsants lamotrigine, riluzole, and valproate differentially regulate AMPA receptor membrane localization: relationship to clinical effects in mood disorders. Neuropsychopharmacology 32:793-802 |
| DU, Jing; Quiroz, Jorge; Yuan, Peixiong et al. (2004) Bipolar disorder: involvement of signaling cascades and AMPA receptor trafficking at synapses. Neuron Glia Biol 1:231-43 |
