Revealing the Landscape of Synaptic Diversity by Cell type- and Synapse-specific...
Revealing the Landscape of Synaptic Diversity by Cell type- and Synapse-specific Proteomics and Transcriptomics
Synapses differ in their shape, size, and function and exhibit ongoing plasticity during the development and modification of neural circuits. The structural diversity of synapses is largely known whereas the molecular diversity of...
Synapses differ in their shape, size, and function and exhibit ongoing plasticity during the development and modification of neural circuits. The structural diversity of synapses is largely known whereas the molecular diversity of synapses is much less well understood. While most synaptic molecules have been identified by immunolabeling and bulk proteomic approaches, the complement of proteins present at individual synapse types, as well as their stoichiometric relationships with other molecules remains unknown. Indeed, our current classification mostly relies on neurotransmitter/receptor phenotypes, leading to broad descriptors like excitatory or inhibitory synapses. Because the function of the synapse, as well as its ability to change, is largely determined by the quality and quantity of molecules that inhabit it, it is essential to understand synaptic molecular diversity.
The aims of this proposal are to determine the proteomes and transcriptomes of genetically-identifiable synaptic populations in different brain areas and to determine the molecular diversity in these same synapse populations using transcriptomic analysis of individual synapses. We will also assess how these synaptic proteomes, transcriptomes and the transcriptomic diversity respond to plasticity.
To study synaptic proteomes and transcriptomes we will use Fluorescence-Activated Synaptosome Sorting to purify different synaptic populations (excitatory, inhibitory, dopaminergic) from different brain areas. We will use the state-of-the-art methods optimized for quantitative transcriptomic and proteomic profiling of very small amounts of sorted synapses and develop a method, SynDrops, for the transcriptomic analysis of individual synapses. We will examine how synaptic proteomes and transcriptomes change during plasticity. These studies will reveal the landscape of synaptic diversity within synapse types and across brain areas and allow the field to probe diseased synapses in the future.ver más
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