Confocal calcium imaging and electrophysiology in live brain cells for neuroscience research at Université de Sherbrooke

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Our research focuses on the role of brainstem and spinal cells in locomotor control (Dr. Ryczko), the role of G-protein coupled receptors (GPCRs) in pain modulation (Dr. Sarret) and the role of delta opioid receptors (DORs) in the olfactory bulbs (Dr. Gendron). The proposal aims at upgrading two setups for calcium imaging and patch-clamp recordings in live brain preparations. The requested equipment will be the core of a new electrophysiology/calcium imaging platform for neuroscience studies at Université de Sherbrooke (UdeS).******Our current equipment (setup1) comprises an epifluorescent microscope for calcium imaging and an electrophysiology rig for patch-clamp recordings. We propose to upgrade this microscope to confocal technology. This requires a laser light source and a confocal unit with computer and software. With our current epifluorescent microscope, light scattering prevents the acquisition of fluorescent signals from cells located deep in the brain tissue. By removing out-of-focus light, confocal technology will allow us to record easily the activity of brain cells at different depths using fluorescent calcium sensors with exquisite resolution, and to perform precise optogenetic stimulations. This will allow us to identify important mechanisms underlying the recruitment and coordination of brain cell activities. Our patch-clamp recordings will also benefit from this upgrade. We will then target the fluorescent cells of interest with patch micropipettes at tissue depths that we cannot reach with our current system. In addition, we will recycle the main items from setup1 (high speed camera, LED light source, computer and software) to build a second epifluorescent calcium imaging/patch-clamp setup (setup2), which will be ideally suited for recordings in mouse brain slices. ***The requested equipment will double our capacity for such recordings and bring to a new level our NSERC research programs by enabling us to address precise pharmacological and physiological mechanistic questions in neuroscience. By having access to the deep layers of the brain and spinal cord, Dr. Ryczko will decode the principles underlying cell recruitment in the locomotor system of the salamander, an animal that can fully repair its nervous system after a lesion. Dr. Sarret will develop the use of patch-clamp electrophysiology and calcium imaging and this will fasten further his discoveries on the role of GPCRs in pain neural networks. Dr. Gendron will gain access for the first time to the cellular activities evoked by compounds acting on DORs in the olfactory bulbs of mammals. ***The setup will bring to our institution the first comprehensive platform combining confocal microscopy, calcium imaging and patch-clamp electrophysiology, providing hands-on training to the students with state-of-the-art neuroscience techniques.**