Two-Photon Imaging of Neural Activity in Awake, Head-Restrained Mice

Wienisch, Martin; Blauvelt, David; Sato, Tomokazu; Murthy, Venkatesh

doi:10.1007/7657_2011_18

Cited by 12 publications

(10 citation statements)

References 33 publications

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“…Mice were trained to tolerate the head-fixed setup (Wienisch et al, 2012), but odors were not associated with a task or reward. Anesthetized mice displayed slow and regular respiration patterns accompanied by individually discernible fluorescence changes (Figure 1C).…”

Section: Resultsmentioning

confidence: 99%

“…After thorough cleaning and drying of the skull, a thin layer of cyanoacrylic adhesive (Vetbond, 3M) was applied to the skull and surrounding skin. Finally, a custom cut titanium headplate (Wienisch et al, 2012) was attached using acrylic cement (Jet Repair, Lang Dental). After the acrylic adhesive hardened (approximately 20 min), the sniff cannula implantation surgery was started.…”

Section: Methodsmentioning

confidence: 99%

“…The center of the wheel was pierced with a 2 mm diameter steel rod acting as an axle, and this axle was mounted to posts damped by soft springs. This method offered the animals' forward and backward freedom of motion while preventing uncontrolled lateral motion that a styrofoam ball may have (Wienisch et al, 2012). Training was started 48 h after headplate implantation surgery.…”

Section: Methodsmentioning

confidence: 99%

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Distinct spatiotemporal activity in principal neurons of the mouse olfactory bulb in anesthetized and awake states

Blauvelt¹,

Sato²,

Wienisch³

et al. 2013

Front. Neural Circuits

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The acquisition of olfactory information and its early processing in mammals are modulated by brain states through sniffing behavior and neural feedback. We imaged the spatiotemporal pattern of odor-evoked activity in a population of output neurons (mitral/tufted cells, MTCs) in the olfactory bulb (OB) of head-restrained mice expressing a genetically-encoded calcium indicator. The temporal dynamics of MTC population activity were relatively simple in anesthetized animals, but were highly variable in awake animals. However, the apparently irregular activity in awake animals could be predicted well using sniff timing measured externally, or inferred through fluctuations in the global responses of MTC population even without explicit knowledge of sniff times. The overall spatial pattern of activity was conserved across states, but odor responses had a diffuse spatial component in anesthetized mice that was less prominent during wakefulness. Multi-photon microscopy indicated that MTC lateral dendrites were the likely source of spatially disperse responses in the anesthetized animal. Our data demonstrate that the temporal and spatial dynamics of MTCs can be significantly modulated by behavioral state, and that the ensemble activity of MTCs can provide information about sniff timing to downstream circuits to help decode odor responses.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Distinct spatiotemporal activity in principal neurons of the mouse olfactory bulb in anesthetized and awake states

Blauvelt¹,

Sato²,

Wienisch³

et al. 2013

Front. Neural Circuits

Self Cite

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show abstract

“…For unanesthetized experiments, a titanium headplate, 37.3×5.2×0.9 mm (L×W×H) weighing 0.5 g, was first affixed to the skull under anesthesia (see Wienisch et al 2011). The dorsal surface of the skull was exposed, a thin layer of cyanoacrylate (VetBond, WPI, Inc.) applied, and the headplate affixed to the dorsal aspect of the skull near bregma with acrylic bonding material (C&B Metabond).…”

Section: Animal Preparationmentioning

confidence: 99%

Sound-Evoked Olivocochlear Activation in Unanesthetized Mice

Chambers

Hancock

Maison

et al. 2011

JARO

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Genetic tools available for the mouse make it a powerful model to study the modulation of cochlear function by descending control systems. Suppression of distortion product otoacoustic emission (DPOAE) amplitude by contralateral acoustic stimulation (CAS) provides a robust tool for noninvasively monitoring the strength of descending modulation, yet investigations in mice have been performed infrequently and only under anesthesia, a condition likely to reduce olivocochlear activation. Here, we characterize the contralateral olivocochlear reflex in the alert, unanesthetized mouse. Head-fixed mice were restrained between two closed acoustic systems, while an artifact rejection protocol minimized contamination from self-generated sounds and movements. In mice anesthetized with pentobarbital, ketamine or urethane, CAS at 80 dB SPL evoked, on average, a G1-dB change in DPOAE amplitude. In contrast, the mean CASinduced DPOAE suppression in unanesthetized mice was nearly 8 dB. Experiments in mice with targeted deletion of the α9 subunit of the nicotinic acetylcholine receptor confirmed the contribution of the medial olivocochlear efferents to this phenomenon. These findings demonstrate the utility of the CAS assay in the unanesthetized mouse and highlight the adverse effects of anesthesia when probing the functional status of descending control pathways within the auditory system.

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“…When combined with surgical preparations [5][6][7] in which a single procedure allows optical access to the brain for weeks to months, these microscopy approaches have been used to study dynamic processes in the brain in healthy and diseased states [8][9][10][11] . In addition, protocols have been developed 12,13 that provide for in vivo imaging in awake (i.e., non anesthetized) animals, allowing for cellular-resolution functional imaging during behavioral assays. These protocols have been used for comparisons of correlated neuronal activity 14 , astrocyte calcium signaling 15 in anesthetized and awake animals, the identification of task-specific neuronal clusters 16 , and the ability of neurons to discriminate object location upon whisker stimulation 17 .…”

Section: Introductionmentioning

confidence: 99%

A Procedure for Implanting a Spinal Chamber for Longitudinal <em>In Vivo </em>Imaging of the Mouse Spinal Cord

Farrar

Schaffer

2014

JoVE

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Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

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Two-Photon Imaging of Neural Activity in Awake, Head-Restrained Mice

Cited by 12 publications

References 33 publications

Distinct spatiotemporal activity in principal neurons of the mouse olfactory bulb in anesthetized and awake states

Distinct spatiotemporal activity in principal neurons of the mouse olfactory bulb in anesthetized and awake states

Sound-Evoked Olivocochlear Activation in Unanesthetized Mice

A Procedure for Implanting a Spinal Chamber for Longitudinal <em>In Vivo </em>Imaging of the Mouse Spinal Cord

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