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1866 Janelia Publications

Showing 31-40 of 1866 results
11/27/20 | Super-resolution ophthalmoscopy: Virtually structured detection for resolution improvement in retinal imaging.
Yao X, Lu R, Wang B, Lu Y, Kim T
Experimental Biology and Medicine (Maywood). 2020 Nov 27:1535370220970533. doi: 10.1177/1535370220970533

Quantitative retinal imaging is essential for advanced study and clinical management of eye diseases. However, spatial resolution of retinal imaging has been limited due to available numerical aperture and optical aberration of the ocular optics. Structured illumination microscopy has been established to break the diffraction-limit resolution in conventional light microscopy. However, practical implementation of structured illumination microscopy for ophthalmoscopy of the retina is challenging due to inevitable eye movements that can produce phase artifacts. Recently, we have demonstrated the feasibility of using virtually structured detection as one alternative to structured illumination microscopy for super-resolution imaging. By providing the flexibility of digital compensation of eye movements, the virtually structured detection provides a feasible, phase-artifact-free strategy to achieve super-resolution ophthalmoscopy. In this article, we summarize the technical rationale of virtually structured detection, and its implementations for super-resolution imaging of freshly isolated retinas, intact animals, and awake human subjects.

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11/26/20 | An adaptive optics module for deep tissue multiphoton imaging in vivo
Cristina Rodríguez , Anderson Chen , José A. Rivera , Manuel A. Mohr , Yajie liang , Wenzhi Sun , Daniel E. Milkie , Thomas G. Bifano , Xiaoke Chen , Na Ji
bioRxiv. 2020 Nov 26:. doi: https://doi.org/10.1101/2020.11.25.397968

Understanding complex biological systems requires visualizing structures and processes deep within living organisms. We developed a compact adaptive optics module and incorporated it into two- and three-photon fluorescence microscopes, to measure and correct tissue-induced aberrations. We resolved synaptic structures in deep cortical and subcortical areas of the mouse brain, and demonstrated high-resolution imaging of neuronal structures and somatosensory-evoked calcium responses in the mouse spinal cord at unprecedented depths in vivo.

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11/26/20 | The art of lineage tracing: from worm to human.
Garcia-Marques J, Espinosa-Medina I, Lee T
Progress in Neurobiology. 2020 Nov 26:101966. doi: 10.1016/j.pneurobio.2020.101966

Reconstructing the genealogy of every cell that makes up an organism remains a long-standing challenge in developmental biology. Besides its relevance for understanding the mechanisms underlying normal and pathological development, resolving the lineage origin of cell types will be crucial to create these types on-demand. Multiple strategies have been deployed towards the problem of lineage tracing, ranging from direct observation to sophisticated genetic approaches. Here we discuss the achievements and limitations of past and current technology. Finally, we speculate about the future of lineage tracing and how to reach the next milestones in the field.

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11/25/20 | Neural circuit mechanisms of sexual receptivity in Drosophila females.
Wang K, Wang F, Forknall N, Yang T, Patrick C, Parekh R, Dickson BJ
Nature. 2020 Nov 25:. doi: 10.1038/s41586-020-2972-7

Choosing a mate is one of the most consequential decisions a female will make during her lifetime. A female fly signals her willingness to mate by opening her vaginal plates, allowing a courting male to copulate. Vaginal plate opening (VPO) occurs in response to the male courtship song and is dependent on the mating status of the female. How these exteroceptive (song) and interoceptive (mating status) inputs are integrated to regulate VPO remains unknown. Here we characterize the neural circuitry that implements mating decisions in the brain of female Drosophila melanogaster. We show that VPO is controlled by a pair of female-specific descending neurons (vpoDNs). The vpoDNs receive excitatory input from auditory neurons (vpoENs), which are tuned to specific features of the D. melanogaster song, and from pC1 neurons, which encode the mating status of the female. The song responses of vpoDNs, but not vpoENs, are attenuated upon mating, accounting for the reduced receptivity of mated females. This modulation is mediated by pC1 neurons. The vpoDNs thus directly integrate the external and internal signals that control the mating decisions of Drosophila females.

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11/25/20 | Theoretical principles for illuminating sensorimotor processing with brain-wide neuronal recordings.
Biswas T, Bishop WE, Fitzgerald JE
Current Opinion in Neurobiology. 2020 Nov 25;65:138-145. doi: 10.1016/j.conb.2020.10.021

Modern recording techniques now permit brain-wide sensorimotor circuits to be observed at single neuron resolution in small animals. Extracting theoretical understanding from these recordings requires principles that organize findings and guide future experiments. Here we review theoretical principles that shed light onto brain-wide sensorimotor processing. We begin with an analogy that conceptualizes principles as streetlamps that illuminate the empirical terrain, and we illustrate the analogy by showing how two familiar principles apply in new ways to brain-wide phenomena. We then focus the bulk of the review on describing three more principles that have wide utility for mapping brain-wide neural activity, making testable predictions from highly parameterized mechanistic models, and investigating the computational determinants of neuronal response patterns across the brain.

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11/24/20 | Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging.
Qian Y, Cosio DM, Piatkevich KD, Aufmkolk S, Su W, Celiker OT, Schohl A, Murdock MH, Aggarwal A, Chang Y, Wiseman PW, Ruthazer ES, Boyden ES, Campbell RE
PLoS Biology. 2020 Nov 24;18(11):e3000965. doi: 10.1371/journal.pbio.3000965

Near-infrared (NIR) genetically encoded calcium ion (Ca2+) indicators (GECIs) can provide advantages over visible wavelength fluorescent GECIs in terms of reduced phototoxicity, minimal spectral cross talk with visible light excitable optogenetic tools and fluorescent probes, and decreased scattering and absorption in mammalian tissues. Our previously reported NIR GECI, NIR-GECO1, has these advantages but also has several disadvantages including lower brightness and limited fluorescence response compared to state-of-the-art visible wavelength GECIs, when used for imaging of neuronal activity. Here, we report 2 improved NIR GECI variants, designated NIR-GECO2 and NIR-GECO2G, derived from NIR-GECO1. We characterized the performance of the new NIR GECIs in cultured cells, acute mouse brain slices, and Caenorhabditis elegans and Xenopus laevis in vivo. Our results demonstrate that NIR-GECO2 and NIR-GECO2G provide substantial improvements over NIR-GECO1 for imaging of neuronal Ca2+ dynamics.

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11/18/20 | Spatial readout of visual looming in the central brain of Drosophila.
Morimoto MM, Nern A, Zhao A, Rogers EM, Wong A, Isaacson MD, Bock D, Rubin GM, Reiser MB
eLife. 2020 Nov 18;9:. doi: 10.7554/eLife.57685

Visual systems can exploit spatial correlations in the visual scene by using retinotopy. However, retinotopy is often lost, such as when visual pathways are integrated with other sensory modalities. How is spatial information processed outside of strictly visual brain areas? Here, we focused on visual looming responsive LC6 cells in , a population whose dendrites collectively cover the visual field, but whose axons form a single glomerulus-a structure without obvious retinotopic organization-in the central brain. We identified multiple cell types downstream of LC6 in the glomerulus and found that they more strongly respond to looming in different portions of the visual field, unexpectedly preserving spatial information. Through EM reconstruction of all LC6 synaptic inputs to the glomerulus, we found that LC6 and downstream cell types form circuits within the glomerulus that enable spatial readout of visual features and contralateral suppression-mechanisms that transform visual information for behavioral control.

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11/17/20 | Multi-regional circuits underlying visually guided decision-making in Drosophila.
Cheong H, Siwanowicz I, Card GM
Current Opinion in Neurobiology. 2020 Nov 17;65:77-87. doi: 10.1016/j.conb.2020.10.010

Visually guided decision-making requires integration of information from distributed brain areas, necessitating a brain-wide approach to examine its neural mechanisms. New tools in Drosophila melanogaster enable circuits spanning the brain to be charted with single cell-type resolution. Here, we highlight recent advances uncovering the computations and circuits that transform and integrate visual information across the brain to make behavioral choices. Visual information flows from the optic lobes to three primary central brain regions: a sensorimotor mapping area and two 'higher' centers for memory or spatial orientation. Rapid decision-making during predator evasion emerges from the spike timing dynamics in parallel sensorimotor cascades. Goal-directed decisions may occur through memory, navigation and valence processing in the central complex and mushroom bodies.

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12/01/20 | Linking axon morphology to gene expression: a strategy for neuronal cell-type classification.
Winnubst J, Spruston N, Harris JA
Current Opinion in Neurobiology. 2020 Dec 01;65:70-76. doi: 10.1016/j.conb.2020.10.006

To study how the brain drives cognition and behavior we need to understand its cellular composition. Advances in single-cell transcriptomics have revolutionized our ability to characterize neuronal diversity. To arrive at meaningful descriptions of cell types, however, gene expression must be linked to structural and functional properties. Axonal projection patterns are an appropriate measure, as they are diverse, change only gradually over time, and they influence and constrain circuit function. Here, we consider how efforts to map transcriptional and morphological diversity in the mouse brain could be linked to generate a modern taxonomy of the mouse brain.

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11/11/20 | Optimized Vivid-derived Magnets photodimerizers for subcellular optogenetics in mammalian cells.
Benedetti L, Marvin JS, Falahati H, Guillén-Samander A, Looger LL, De Camilli P
Elife. 2020 Nov 11;9:. doi: 10.7554/eLife.63230

Light-inducible dimerization protein modules enable precise temporal and spatial control of biological processes in non-invasive fashion. Among them, Magnets are small modules engineered from the photoreceptor Vivid by orthogonalizing the homodimerization interface into complementary heterodimers. Both Magnets components, which are well-tolerated as protein fusion partners, are photoreceptors requiring simultaneous photoactivation to interact, enabling high spatiotemporal confinement of dimerization with a single-excitation wavelength. However, Magnets require concatemerization for efficient responses and cell preincubation at 28C to be functional. Here we overcome these limitations by engineering an optimized Magnets pair requiring neither concatemerization nor low temperature preincubation. We validated these 'enhanced' Magnets (eMags) by using them to rapidly and reversibly recruit proteins to subcellular organelles, to induce organelle contacts, and to reconstitute OSBP-VAP ER-Golgi tethering implicated in phosphatidylinositol-4-phosphate transport and metabolism. eMags represent a very effective tool to optogenetically manipulate physiological processes over whole cells or in small subcellular volumes.

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