Synaptic connections in the brain are continuously remodeled in response to neuronal activity. This process, known as synaptic plasticity, is widely accepted as the cellular basis for learning and memory, and it is thought to be altered in several cognitive disorders. An important aspect of synaptic plasticity is the regulated transport of neurotransmitter receptors in and out of synapses. In particular, AMPA-type glutamate receptors (AMPA receptors) can be added to or removed from synapses in an activity-dependent manner, leading to long-lasting changes in synaptic function (long-term potentiation and long-term depression). These forms of synaptic plasticity are widely thought to underlie learning and memory.
An important lag in our understanding of receptor trafficking as an underlying mechanism for synaptic plasticity stems from the fact that receptor delivery at synapses has never been observed in living animals in response to physiological brain activity. In fact, progress in this field is currently hindered by the challenge of visualizing neurotransmitter receptor trafficking in real-time in living brain. This proposal is aimed at directly imaging the synaptic insertion of AMPA receptors in response to neuronal activity in the brain in vivo. Our hypothesis is that physiological sensory stimulation will be able to trigger the mobilization and insertion of AMPA receptors from extrasynaptic dendritic compartments into the synaptic membrane at dendritic spines. To this end we will use two-photon laser scanning fluorescence microscopy combined with expression of GFP-tagged AMPA receptors in the somatosensory cortex of living rats.
Using this combination of molecular biology together with cutting-edge fluorescence microscopy, we will determine (i) the basal dynamics of AMPA receptors at dendritic spines in the intact brain, (ii) the regulated delivery of AMPA receptors at synapses in response to sensory stimulation, and (iii) the signaling cascades that mediate the activity-dependent trafficking of AMPA receptors in vivo.
We believe that this molecular imaging approach in intact living brain is essential for the general goal of understanding how individual molecules contribute to normal brain function and to the pathological alterations associated with mental illness.