Probing Synaptic Signaling with Optogenetic Stimulation and Genetically Encoded Calcium Reporters
- 1 September 2020
- book chapter
- protocol
- Published by Springer Science and Business Media LLC
- Vol. 2191, 109-134
- https://doi.org/10.1007/978-1-0716-0830-2_8
Abstract
Optogenetics provides a powerful approach for investigating neuronal electrophysiology at the scale required for drug discovery applications. Probing synaptic function with high throughput using optogenetics requires robust tools that enable both precise stimulation of and facile readout of synaptic activity. Here we describe two functional assays to achieve this end: (1) a pre-synaptic calcium assay that utilizes the channelrhodopsin, CheRiff, patterned optogenetic stimulus, and the pre-synaptically targeted calcium reporter jRGECO1a to monitor pre-synaptic changes in calcium influx and (2) a synaptic transmission assay in which CheRiff and cytosolic jRGECO1a are expressed in non-overlapping sets of neurons, enabling pre-synaptic stimulation and post-synaptic readout of activity. This chapter describes the methodology and practical considerations for implementation of these two assays.Keywords
This publication has 37 references indexed in Scilit:
- De novo mutations in the gene encoding the synaptic scaffolding proteinSHANK3in patients ascertained for schizophreniaProceedings of the National Academy of Sciences of the United States of America, 2010
- Presynaptic dysfunction in Huntington's diseaseBiochemical Society Transactions, 2010
- Increased Expression of α-Synuclein Reduces Neurotransmitter Release by Inhibiting Synaptic Vesicle Reclustering after EndocytosisNeuron, 2010
- Amyloid-β as a positive endogenous regulator of release probability at hippocampal synapsesNature Neuroscience, 2009
- Analysis of gene expression in two large schizophrenia cohorts identifies multiple changes associated with nerve terminal functionMolecular Psychiatry, 2009
- Genome-wide association for major depressive disorder: a possible role for the presynaptic protein piccoloMolecular Psychiatry, 2008
- Genetic and expression analyses reveal elevated expression of syntaxin 1A ( STX1A) in high functioning autismInternational Journal of Neuropsychopharmacology, 2008
- De novo mutations in the gene encoding STXBP1 (MUNC18-1) cause early infantile epileptic encephalopathyNature Genetics, 2008
- Mutations in the gene encoding the synaptic scaffolding protein SHANK3 are associated with autism spectrum disordersNature Genetics, 2006
- Synaptosomal-associated protein 25 (SNAP-25) and attention deficit hyperactivity disorder (ADHD): evidence of linkage and association in the Irish populationMolecular Psychiatry, 2002