Trans-segmental imaging in the spinal cord of behaving mice

Shekhtmeyster, Pavel; Duarte, Daniela; Carey, Erin M.; Ngo, Alexander; Gao, Grace; Olmstead, Jack A; Nelson, Nicholas A.; Nimmerjahn, Axel

doi:10.1038/s41587-023-01700-3

Cited by 14 publications

(13 citation statements)

References 24 publications

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“…We employed AQuA2 to analyze translaminar astrocyte calcium activity in the spinal dorsal horn of Tg(GFAP:GCaMP6f) mice. The experimental setup, depicted in Figure 5A, involved securing the mouse on a spherical treadmill and optically recording GCaMP6f activity through a microprism implanted near the lateral edge of the spinal gray matter 8 . During the recordings, a pressure stimulus was applied to the mouse’s proximal tail.…”

Section: Resultsmentioning

confidence: 99%

“…AQuA2 can detect microglial calcium activity, as shown in Figure 6A. These transients were recorded in the superficial dorsal horn of behaving Tg(CX3CR1:GCaMP5g-tdTomato) mice using a wearable macroscope 46 . Both somatic signals and signals with irregular shapes and complex spatiotemporal features were identified.…”

Section: Aqua2 Quantifies Signals Across Biosensors Cell Types Organs...mentioning

confidence: 99%

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Fast, Accurate, and Versatile Data Analysis Platform for the Quantification of Molecular Spatiotemporal Signals

Mi,

Chen,

Duarte

et al. 2024

Preprint

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Optical recording of intricate molecular dynamics is becoming an indispensable technique for biological studies, accelerated by the development of new or improved biosensors and microscopy technology. This creates major computational challenges to extract and quantify biologically meaningful patterns embedded within complex and rich data sources. Here, we introduce Activity Quantification and Analysis (AQuA2), a fast, accurate, and versatile data analysis platform built upon advanced machine learning techniques. AQuA2 decomposes complex live imaging-based datasets into elementary signaling events, allowing accurate and unbiased quantification of molecular activities and identification of consensus functional units. We demonstrate applications of AQuA2 across a range of biosensors (calcium, norepinephrine, ATP, acetylcholine, dopamine), cell types (astrocytes, oligodendrocytes, microglia, neurons), organs (brains and spinal cords), animal models (zebrafish and mouse), and imaging modalities (confocal, two-photon, light sheet). As exemplar findings, we show how AQuA2 identified drug-dependent interactions between neurons and astroglia, and distinct sensorimotor signal propagation patterns in the mouse spinal cord.

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Section: Resultsmentioning

confidence: 99%

Section: Aqua2 Quantifies Signals Across Biosensors Cell Types Organs...mentioning

confidence: 99%

Fast, Accurate, and Versatile Data Analysis Platform for the Quantification of Molecular Spatiotemporal Signals

Mi,

Chen,

Duarte

et al. 2024

Preprint

Self Cite

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show abstract

“…For example, spinal cord flexibility during natural locomotion, exacerbated by breathing, produces motion artifacts in imaging data and hinders quantitative analysis. Not surprisingly, few studies have recorded spinal cord neural activity, long-term in awake animals (Cheng et al, 2016;Sekiguchi et al, 2016;Ju et al, 2022;Shekhtmeyster et al, 2023).…”

Section: Main Textmentioning

confidence: 99%

Long-term optical imaging of the spinal cord in awake, behaving animals

Ahanonu

Crowther

Kania

et al. 2023

Preprint

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Advances in optical imaging approaches and fluorescent biosensors have enabled an understanding of the spatiotemporal and long-term neural dynamics in the brain of awake animals. However, methodological difficulties and the persistence of post-laminectomy fibrosis have greatly limited similar advances in the spinal cord. To overcome these technical obstacles, we combinedin vivoapplication of fluoropolymer membranes that inhibit fibrosis; a redesigned, cost-effective implantable spinal imaging chamber; and improved motion correction methods that together permit imaging of the spinal cord in awake, behaving mice, for months to over a year. We also demonstrate a robust ability to monitor axons, identify a spinal cord somatotopic map, conduct Ca2+imaging of neural dynamics in behaving animals responding to pain-provoking stimuli, and observe persistent microglial changes after nerve injury. The ability to couple neural activity and behavior at the spinal cord level will drive insights not previously possible at a key location for somatosensory transmission to the brain.

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“…Among other behaviorally relevant state changes, astrocytes have been implicated in controlling sleep homeostasis in multiple species (Blum et al, 2021;Clasadonte et al, 2017;Ingiosi and Frank, 2022). Astrocyte calcium levels are thought to be the primary means through which these cells accumulate information about the circadian cycle, a hypothesis supported by pioneering imaging studies in unrestrained mice using head-mounted miniscopes (Ingiosi et al, 2020;Kjaerby et al, 2020;Shekhtmeyster et al, 2021). By implementing our multi-core optical fiber imaging system, we were able to extend this analysis over 24hr imaging periods to resolve longitudinal activity patterns of individual astrocytes within the cerebral cortex, providing direct evidence that calcium activity within individual astrocytes varies prominently with the internal clock of the animals.…”

Section: Astrocytes Exhibit Distinct Calcium Activity Patterns During...mentioning

confidence: 99%

Multicore fiber optic imaging reveals that astrocyte calcium activity in the cerebral cortex is modulated by internal motivational state

Gau

Hsu

Cha

et al. 2023

Preprint

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Astrocytes are a direct target of neuromodulators and can influence neuronal activity on broad spatial and temporal scales through their close proximity to synapses. However, our knowledge about how astrocytes are functionally recruited during different animal behaviors and their diverse effects on the CNS remains limited. To enable measurement of astrocyte activity patterns in vivo during normative behaviors, we developed a high-resolution, long working distance, multi-core fiber optic imaging platform that allows visualization of cortical astrocyte calcium transients through a cranial window in freely moving mice. Using this platform, we defined the spatiotemporal dynamics of astrocytes during diverse behaviors, ranging from circadian fluctuations to novelty exploration, showing that astrocyte activity patterns are more variable and less synchronous than apparent in head-immobilized imaging conditions. Although the activity of astrocytes in visual cortex was highly synchronized during quiescence to arousal transitions, individual astrocytes often exhibited distinct thresholds and activity patterns during explorative behaviors, in accordance with their molecular diversity, allowing temporal sequencing across the astrocyte network. Imaging astrocyte activity during self-initiated behaviors revealed that noradrenergic and cholinergic systems act synergistically to recruit astrocytes during state transitions associated with arousal and attention, which was profoundly modulated by internal state. The distinct activity patterns exhibited by astrocytes in the cerebral cortex may provide a means to vary their neuromodulatory influence in response to different behaviors and internal states.

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Trans-segmental imaging in the spinal cord of behaving mice

Cited by 14 publications

References 24 publications

Fast, Accurate, and Versatile Data Analysis Platform for the Quantification of Molecular Spatiotemporal Signals

Fast, Accurate, and Versatile Data Analysis Platform for the Quantification of Molecular Spatiotemporal Signals

Long-term optical imaging of the spinal cord in awake, behaving animals

Multicore fiber optic imaging reveals that astrocyte calcium activity in the cerebral cortex is modulated by internal motivational state

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