Experience-dependent alterations in inhibition using high-throughput and input-specific fluorescence synapse imaging

• Autorzy: Kuljis D.A. , Matsushita M.T. , Park E. , Telmer C.A. , Bruchez M.P. , Barth A.L.
• Języki publikacji: EN

Abstrakty

EN

Fluorescence-based synapse detection enables dense, high-throughput and multichannel analysis of synapse properties and connections in brain tissue. Using fluorogen activating proteins (FAPs) or YFP coupled to a neuroligin tether, we have developed genetically-encoded reagents for fluorescence-labeling of post-synaptic sites. Sparse viral expression of YFP-post or FAP-post in mouse somatosensory (barrel) cortex enables compartment-specific quantitation of synapses across the surface of an individual neuron, as well as cell-type specific presynaptic input assignment. High-resolution, 3D confocal stacks were used for semi-automated, high-throughput assignment of YFP-post and FAP-post synaptic puncta across specific neuron cell-types. Using transgenic mice where specific subtypes of presynaptic inhibitory neurons were fluorescently-labeled with YFP, far red-fluorescence of FAP-post synaptic puncta and dTomato-filled postsynaptic pyramidal cells could be aligned to specific presynaptic partners using tricolor colocalization. Fluorescent post-synaptic puncta properties were evaluated to generate metrics reflecting synapse location, size, shape, and fluorescence intensity that could be used to differentiate synapses from different inhibitory sources. We used these quantitative metrics to evaluate changes in inhibitory inputs after sensory association training for pyramidal neurons in barrel cortex. Genetically-encoded fluorescence-based synaptic labeling reagents provide a powerful approach to enable high-throughput and automated analysis of synapse organization in brain tissue across development, learning, and disease states.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2017
• Tom: 77
• Numer: Suppl.1
• Opis fizyczny: p.37

Twórcy

autor

Kuljis D.A.

• Biological Sciences Department, Carnegie Mellon University, Pittsburgh, USA
• Neurobiology Department, University of Pittsburgh, Pittsburgh, USA
• Center for the Neural Basis of Cognition, University of Pittsburgh, Pittsburgh, USA

autor

Matsushita M.T.

• Biological Sciences Department, Carnegie Mellon University, Pittsburgh, USA

autor

Park E.

• Biological Sciences Department, Carnegie Mellon University, Pittsburgh, USA

autor

Telmer C.A.

• Department of Chemistry, Carnegie Mellon University, Pittsburgh, USA
• Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, USA

autor

Bruchez M.P.

• Biological Sciences Department, Carnegie Mellon University, Pittsburgh, USA
• Department of Chemistry, Carnegie Mellon University, Pittsburgh, USA
• Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, USA

autor

Barth A.L.

• Biological Sciences Department, Carnegie Mellon University, Pittsburgh, USA
• Center for the Neural Basis of Cognition, University of Pittsburgh, Pittsburgh, USA