Spatio-temporally precise manipulation and read-out of brain circuit function has been one of the longest-standing challenges in neuroscience. The recent explosion in the field of genetically encoded tools to control and measure neuronal activity has greatly facilitated investigation of brain function, ranging from single synapses to large-scale circuits. Both control and readout of neuronal activity can now be achieved over orders of magnitude in space and time, ranging from micrometers to entire brain regions and from milliseconds to days.
This course will provide participants with the opportunity to gain hands-on experience using the latest genetically encoded tools and state-of-the-art equipment for brain circuit investigation. A particular focus will lie on multiplexed manipulations and read-out of brain circuits. Participants will be familiarized with the biophysical principles behind the sensors and actuators, and given training complementary to their background in the technical aspects of experimental approaches.
Hands-on experiments will employ optogenetic and chemogenetic actuators, including excitatory and inhibitory ion channels, pumps, enzymes and G-protein coupled receptors. These actuators will be complemented by genetically encoded indicators of neural activity, including calcium and voltage indicators as well as indicators for neurotransmitters and neuromodulators such as glutamate, dopamine and norepinephrine.
The course will cover a wide range of experimental systems with an emphasis on functional brain circuits in vivo. Finally, participants will be guided through data analysis and conceptual interpretations of their experiments.
Course director & co-directors
- Ofer Yizhar (Weizmann Institute of Science, Israel)
- Michael Lin (Stanford University, USA)
- Simon Wiegert (Center for Molecular Neurobiology Hamburg, Germany)
- Anna Beyeler (Bordeaux Neurocampus, France)
Kris Blanchard (Institut de la vision, France)
Alexander Dieter (ZMNH Germany)
Lief Fenno (University of Stanford, USA)
Nitzan Geva (Weizmann Institute of science, Israel)
Nikolas Karalis (Friedrich Miescher Institute of Basel, Switzerland)
Marie Labouesse (ETH Zurich, Switzerland)
Mathias Mahn (Friedrich Miescher Institute of Basel, Switzerland)
Vasyl Mykytiuk (Max Plank Institute, Germany)
Praneeth Namburi (Institut Magendie, France)
Mauro Pulin (ZMNH, Germany)
Robson Scheffer Teixeira (Max Plank Institute, Germany)
Tristan Schuman (Mount Sinai Institute, New York, USA)
Guilherme Silva (University of Harvard, USA)
Dimitrii Tanese (Institut de la vision, France)
Meytar Zemer (Weizmann Institute of science, Israel)
The following techniques will be covered during the course:
- Implantation of optical fibers, stereotactic injection of viruses in mice.
- Optogenetic stimulation (BIPOLES) and monitoring of pupil size and mouse behavior.
- Imaging of calcium responses, building of a reward delivery system and a head-fixation system for mice either immobilized in a tube or running on a treadmill.
- In vitro exploration of FLARE technique
- 2-photon holographic illumination to achieve single-cell resolved optogenetic activation of presynaptic cells and patch-clamp recording of the post-synaptic neuron
- Concurrent photostimulation and calcium imaging of the presynaptic cells, and voltage imaging on the post-synaptic cell
- Use of miniature head-mounted microscopes (Inscopix).
- Perform population level data analysis on data that was collected across multiple days
- Behavioral assays including the elevated plus maze, the open field test, consumption of water, sucrose, quinine and food, as well as mild foot shocks
- Head-bar implantation, stereotaxic intracerebral virus injection, and craniotomy preparation for long-term recordings
- Head-fixed electrophysiology setup building
- Silicon-probe recordings, in head-fixed mice during behavior, using multi-shank, high-density silicon probes (128-512 channels) & Physiological monitoring
- Opto-tagging and optogenetic manipulations of specific cell-types.
- Open-source hardware and software for the acquisition and processing of the data, including OpenEphys, Bonsai, PulsePal, Cyclops, Arduino, Linux, KiloSort, and MountainSort
- Construct microdrives, mount the silicon probes onto them, make optical fibers of custom length
- Project 1: Optogenetic control of neuromodulation
- Project 2: Optical tool exploration in culture, in slice, and in vivo
- Project 3: Optical tool exploration in culture, in slice, and in vivo
- Project 4: Probing neuronal excitability with Arch-derived voltage sensors
- Project 5: Longitudinal calcium imaging in freely behaving mice using Inscopix system
- Project 6: In vivo imaging of divergent neural populations using dual-color fiber photometry
- Project 7: All optical characterization of eOPN3 mediated terminal inhibition in vivo
- Project 8: Large-scale electrophysiology and optogenetics during head-fixed behavior
- Project 9: In vivo calcium imaging with open-source Miniscopes
- Project 10: Combining in vivo electrophysiology and optogenetics in freely moving mice
- Project 11: All optical interrogation of dopamine circuits in freely moving mice using multiplexfiber photometry and biosensors
- Project 12: All-optical manipulation and read-out of synaptic transmission
Adam Cohen (Harvard University, USA)
Stephane Dieudonne (University of Marseille, France)
Valentina Emiliani (Institut de la vision, France)
Viviana Gradinaru (Caltech applications, USA)
Peter Hegeman (Humboldt University , Germany)
Stefan Herlitze (University of Bochum, Germany
Na Ji (University of Berkeley, USA)
Tom Kash (University of North Carolina, USA)
Sonia Kleinlogel (Bern University, Switzerland)
Tatiana Korotkova (MPI Köln, Germany)
Tommaso Patriarchi (ETH Zurich, Switzerland)
Yaniv Ziv (Weizmann Institute of science, Israel)