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Windows R Us Florence Sc – Eros Quarta, Alessandro Scaglione, Jessica Lucchesi, Leonardo Sacconi, Anna Letizia Allegra Mascaro and Francesco Saverio Pavone

1 Department of Physics and Astronomy, University of Florence, Sesto Fiorentino, Florence, 50019, Italy2 European Laboratory for Nonlinear Spectroscopy, Sesto Fiorentino, Florence, 50019, Italy

Windows R Us Florence Sc

Windows R Us Florence Sc

2 European Laboratory of Nonlinear Spectroscopy, Sesto Fiorentino, Florence, 50019, Italy3 National Institute of Optics, National Research Council, Sesto Fiorentino, Florence, 50019, Italy

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2 European Laboratory for Nonlinear Spectroscopy, Sesto Fiorentino, Florence, 50019, Italy 4 Neuroscience Institute, National Research Council, Pisa, 56124, Italy

1 Department of Physics and Astronomy, University of Florence, Sesto Fiorentino, Florence, 50019, Italy2 European Laboratory of Nonlinear Spectroscopy, Sesto Fiorentino, Florence, 50019, Italy 3 National Institute of Optics, National Research Council, Sesto Fiorentino, 50 Italy

The extent to which features relevant to neocortical processing functions are localized or distributed is a long-standing question in systems neuroscience. Coordinated activity across the neocortex has recently been shown to drive complex behavior in rats, but activity in select regions is canonically associated with specific tasks (eg, movements in the case of the motor cortex). Reaching-to-grasp (RtG) movements are known to depend on neocortical motor circuits; However, the overall activity of the neocortex during these movements remains largely unexplored in rats. Here, using wide-field calcium imaging, we characterized these neocortical-scale dynamics in rats of both sexes engaged in RtG function. Beyond motor areas, we demonstrate that several regions such as the visual and retrosplenial cortices increase their activity levels during successful RtGs, and that homogeneous regions across the ipsilateral hemisphere are also involved. Functional connectivity between neocortical regions transiently increases around movement initiation and decreases during movement. Despite this global phenomenon, neural activity levels correlated with kinematic measures of successful RtGs only in sensitive areas. Our results establish that distributed and local neocortical dynamics co-orchestrate the effective control of complex movements.

A statement of importance depends on mammals’ reaching and grasping movements for small-scale interaction with the physical world. In rats, the motor cortex is critical for performing such behavior, but little is known about patterns of activity in neocortical regions. Using meso-level networks as a model of cortical processing, we investigated the hypothesis that regions beyond motor regions may participate in RtG planning and execution, and that a larger network of regions is involved when performing RtGs. Movement kinematics are mainly correlated with neural activity in sensory areas. By demonstrating that distributed and localized neocortical dynamics co-exist for fine movement execution in the mouse neocortex during RtG, we provide unprecedented insight into the neocortical correlates of mammalian motor control.

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How the CNS generates goal-directed movements, such as reach-to-grasp (RtG), is critical to expanding our understanding of the neural computations that underlie motor ability and animal cognition in general (Bayne et al., 2019). RtG is known to depend on neocortical circuitry, and intensive research has provided detailed information on how neural activity in motor areas of the neocortex contributes to movement control (Evarts, 1968; Georgopoulos et al., 1982; Graziano et al., 2002; Churchland et al., 2012). In rodents, medial prefrontal regions such as the secondary motor cortex (also called medial agranular cortex, precentral cortex, or frontal visual field) have been shown to be necessary for movement preparation (N. Li et al., 2016 Berthas and Kwan, 2017; Chen et al., 2017). Nevertheless, how neural networks produce behavior cannot be fully understood by analyzing its components in isolation. Indeed, in addition to patterns of neural activity at the local level, information processing occurring at the corticocortical level is essential for complex behavior (Peters et al., 2014; Allen et al., 2017; Battaglia-Meyer and Caminin, 2019).

However, despite adoption of the mouse to elucidate the neural basis of motor control (Ölveczky, 2011), neocortex-wide dynamics during RtG remain largely unexplored. Advances in optical methods, particularly wide-field fluorescence microscopy, have made it possible to monitor neuronal activity of almost the entire dorsal neocortex in awake and behaving rats (Allegra Mascaro et al., 2019; Montagni et al., 2019; Sancataldot al., 2019; Ren and Komiyama, 2021 ). With this tool, it has recently been shown that neocortex-scale dynamics emerge during learning (Mackino et al., 2017); Nevertheless, the contribution of areas beyond contralateral motor areas and corticocortical interactions during skilled movements remain largely unexplored. Furthermore, while movement kinematics are associated with neural activity in contralateral motor areas, information about this activity can be used to predict several properties of executed movement (Prsa et al., 2017; C. Li et al., 2019), information on the relationship between RtG kinematics and neural activity across the neocortex is still lacking. is rare.

Our results indicate that global cortical activation emerges during successful RtGs, with functional connectivity (FC) in regions surrounding movement onset, but movement kinematics correlates only with activity in a portion of the sensory region.

Windows R Us Florence Sc

All experimental procedures were authorized by the Italian Ministry of Health (authorization 127-2018-PR). A total of 9 mice (3-11 months of age) were used in this study. Six (3 male, 3 female) GCaMP mice (C57BL/6J-Tg(Thy1-

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Mice (Tg(Thy1-EGFP) MJrs, stock #007788) were used as controls to account for potential hemodynamic artifacts. Animals were kept on a 12 h light/dark cycle

Water Mice undergoing behavioral training were restricted to 80%–90% of their original body weight by restricting food intake to 2–3 g/d. Animals are monitored and weighed daily and feed ratios are increased if necessary. Veterinary staff monitored cattle twice a week.

The day before surgery, mice received a subcutaneous injection of enrofloxacin (10 mg/kg) to prevent infection and then every other day for 3 days. On the day of surgery, animals were anesthetized with isoflurane (3% induction, 1.5% maintenance) and placed on a stereotaxic machine (Kopf Instruments). Absence of tail reflex and toe pinch reflex were tested to confirm that the rat was properly anesthetized. Throughout the operation, the temperature was kept constant at 37°C by an electric heating pad controlled by a rectal temperature sensor (Stolting). To prevent corneal dehydration, ophthalmic gel is applied to the eyes. The scalp was first scrubbed with ethanol and betadine, then a depilatory cream was applied to remove hair, followed by a few drops (2% lidocaine hydrochloride) as an analgesic. A circular piece of scalp was removed and anatomical reference points (bregma and λ) were colored with a permanent marker. Two semicircular cover glasses, one for each hemisphere, were bonded to the skull above the cortex using transparent dental cement. A custom-made headpost was placed ∼0.5 mm back from λ. At the end of surgery, subcutaneous injection of analgesics and anti-inflammatory (carprofen, 5 mg/kg) was administered to facilitate recovery. For this purpose, 200 ml of Ringer’s lactate solution of 0.9% saline was administered to each mouse at the end of surgery. Mice were allowed to recover at least 1 week after surgery. During this period, the animals received

) consisted of a stage (size 300 × 300 × 20 mm, Thorlabs) and walls to form an enclosure and fixed on a laboratory socket (MLJ050/M, Thorlabs) and was inspired by the work of Guo et al. (2015) The enclosure consisted of a turntable attached to a servo motor (MX-28AT) equipped with a non-contact magnetic rotary encoder (MX-28AT) and controlled by a controller board (ArbotiX-M. Robocontroller, Trouser Robotics) to ensure the stable position of the pellets, as well as a base, a perch, a loudspeaker to provide an auditory signal to the animals. The servo is controlled by a temporary program. Chocolate lozenges (10 mg, 5TUL Purified 10 mg Lozenges, Catalog 1811529, TestDiet) were used to induce RtG movements, thus acting as stimuli. The interstimulus interval was 6 to 20 s. The speaker is housed in a custom-made sensory isolating closed imaging recording chamber, which is kept dark and insulated with acoustic foam to further reduce ambient sounds (RS components and woods).

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Behavioral data were collected at 200 fps by a high-speed camera (CM3-U3-13YC-CS, Chameleon3, FLIR) with 2.8-8mm varifocal lenses (LENS-30F2-V80CS, Fujinon ) placed on the left side of the animal. Fixed head. A 470 nm LED illuminating the scene was used as a light source to create a high contrast image for processing and quantification. Movies were recorded using Fly Capture (FLIR) software.

Imaging was performed through the intact skull using a custom-made microscope. The microscope consisted of back-to-back 50 mm f/1.2 camera lenses (Nikon) separated by an FF495-D 03-50.8-D dichroic mirror (Semrac) mounted on a 60 mm cube (ThorLabs). to provoke

Indicator, a 470 nm light source (LED, M470L3, Thorlabs) was applied to an objective (TL2X-SAP 2× super apochromatic microscope objective, 0.61 NA, 0.6.3 NA, LABS LABS 5, 0.6.3mm 5, ) and the fluorescence signal was filtered by a bandpass filter (525 /50 selected by Semrock) and collected by a CMOS camera (ORCA-Flash 4.0 V2, Hamamatsu). Images were acquired at 25 Hz with a resolution of 512 × 512 pixels with a FOV of approximately 12 × 12 mm (16-bit depth) by HCImage Live software (Hamamatsu). Thus the microscope allows an FOV that embraces the entire dorsal view of the rat neocortex. To reduce unnecessary light scatter on the mouse

Windows R Us Florence Sc

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