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3767 Publications

Showing 61-70 of 3767 results
07/22/23 | Dissecting Cell Plate Development During Plant Cytokinesis.
Sinclair R, Cox D, Heddleston J, Aaron J, Wait E, Wilkop T, Drakakaki G
Microscopy and Microanalysis. 2023 Jul 22;29(Supplement_1):865. doi: 10.1093/micmic/ozad067.428
07/22/23 | The density of regulatory information is a major determinant of evolutionary constraint on non-coding DNA in Drosophila
Gonzalo Sabarís , Daniela M. Ortíz , Ian Laiker , Ignacio Mayansky , Sujay Naik , Giacomo Cavalli , David L. Stern , Ella Preger-Ben Noon , Nicolás Frankel
bioRxiv. 2023 Jul 22:. doi: 10.1101/2023.07.21.550043

The density and distribution of regulatory information in non-coding DNA of eukaryotic genomes is largely unknown. Evolutionary analyses have estimated that ∼60% of nucleotides in intergenic regions of the D. melanogaster genome is functionally relevant. This estimate is difficult to reconcile with the commonly accepted idea that enhancers are compact regulatory elements that generally encompass less than 1 kilobase of DNA. Here, we approached this issue through a functional dissection of the regulatory region of the gene shavenbaby (svb). Most of the ∼90 kilobases of this large regulatory region is highly conserved in the genus Drosophila, though characterized enhancers occupy a small fraction of this region. By analyzing the regulation of svb in different contexts of Drosophila development, we found that the regulatory architecture that drives svb expression in the abdominal pupal epidermis is organized in a dramatically different way than the information that drives svb expression in the embryonic epidermis. While in the embryonic epidermis svb is activated by compact and dispersed enhancers, svb expression in the pupal epidermis is driven by large regions with enhancer activity, which occupy a great portion of the svb cis-regulatory DNA. We observed that other developmental genes also display a dense distribution of putative regulatory elements in their regulatory regions. Furthermore, we found that a large percentage of conserved non-coding DNA of the Drosophila genome is contained within putative regulatory DNA. These results suggest that part of the evolutionary constraint on non-coding DNA of Drosophila is explained by the density of regulatory information.

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07/22/23 | Towards Generalizable Organelle Segmentation in Volume Electron Microscopy.
Heinrich L, Patton W, Bennett D, Ackerman D, Park G, Bogovic JA, Eckstein N, Petruncio A, Clements J, Pang S, Shan Xu C, Funke J, Korff W, Hess H, Lippincott-Schwartz J, Saalfeld S, Weigel A, CellMap Project Team
Microscopy and Microanalysis. 2023 Jul 22;29(Supplement_1):975. doi: 10.1093/micmic/ozad067.487
07/21/23 | Nonlinear manifolds underlie neural population activity during behaviour.
Fortunato C, Bennasar-Vázquez J, Park J, Chang JC, Miller LE, Dudman JT, Perich MG, Gallego JA
bioRxiv. 2023 Jul 21:. doi: 10.1101/2023.07.18.549575

There is rich variety in the activity of single neurons recorded during behaviour. Yet, these diverse single neuron responses can be well described by relatively few patterns of neural co-modulation. The study of such low-dimensional structure of neural population activity has provided important insights into how the brain generates behaviour. Virtually all of these studies have used linear dimensionality reduction techniques to estimate these population-wide co-modulation patterns, constraining them to a flat "neural manifold". Here, we hypothesised that since neurons have nonlinear responses and make thousands of distributed and recurrent connections that likely amplify such nonlinearities, neural manifolds should be intrinsically nonlinear. Combining neural population recordings from monkey motor cortex, mouse motor cortex, mouse striatum, and human motor cortex, we show that: 1) neural manifolds are intrinsically nonlinear; 2) the degree of their nonlinearity varies across architecturally distinct brain regions; and 3) manifold nonlinearity becomes more evident during complex tasks that require more varied activity patterns. Simulations using recurrent neural network models confirmed the proposed relationship between circuit connectivity and manifold nonlinearity, including the differences across architecturally distinct regions. Thus, neural manifolds underlying the generation of behaviour are inherently nonlinear, and properly accounting for such nonlinearities will be critical as neuroscientists move towards studying numerous brain regions involved in increasingly complex and naturalistic behaviours.

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07/20/23 | Toward scalable reuse of vEM data: OME-Zarr to the rescue.
Rzepka N, Bogovic JA, Moore JA
Methods in Cell Biology. 2023 Jul 20;177:359-387. doi: 10.1016/bs.mcb.2023.01.016

The growing size of EM volumes is a significant barrier to findable, accessible, interoperable, and reusable (FAIR) sharing. Storage, sharing, visualization and processing are challenging for large datasets. Here we discuss a recent development toward the standardized storage of volume electron microscopy (vEM) data which addresses many of the issues that researchers face. The OME-Zarr format splits data into more manageable, performant chunks enabling streaming-based access, and unifies important metadata such as multiresolution pyramid descriptions. The file format is designed for centralized and remote storage (e.g., cloud storage or file system) and is therefore ideal for sharing large data. By coalescing on a common, community-wide format, these benefits will expand as ever more data is made available to the scientific community.

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07/15/23 | Pinpoint: trajectory planning for multi-probe electrophysiology and injections in an interactive web-based 3D environment
Daniel Birman , Kenneth J. Yang , Steven J. West , Bill Karsh , Yoni Browning , the International Brain Laboratory , Joshua H. Siegle , Nicholas A. Steinmetz
bioRxiv. 2023 Jul 15:. doi: 10.1101/2023.07.14.548952

Targeting deep brain structures during electrophysiology and injections requires intensive training and expertise. Even with experience, researchers often can't be certain that a probe is placed precisely in a target location and this complexity scales with the number of simultaneous probes used in an experiment. Here, we present Pinpoint, open-source software that allows for interactive exploration of stereotaxic insertion plans. Once an insertion plan is created, Pinpoint allows users to save these online and share them with collaborators. 3D modeling tools allow users to explore their insertions alongside rig and implant hardware and ensure plans are physically possible. Probes in Pinpoint can be linked to electronic micro-manipulators allowing real-time visualization of current brain region targets alongside neural data. In addition, Pinpoint can control manipulators to automate and parallelize the insertion process. Compared to previously available software, Pinpoint's easy access through web browsers, extensive features, and real-time experiment integration enable more efficient and reproducible recordings.

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07/13/23 | Localization of fixed dipoles at high precision by accounting for sample drift during illumination
Hinterer F, Schneider MC, Hubmer S, López-Martínez M, Ramlau R, Schütz GJ
Applied Physics Letters. 2023 Jul 13;123(2):. doi: 10.1063/5.0137834

Single molecule localization microscopy relies on the precise quantification of the position of single dye emitters in a sample. This precision is improved by the number of photons that can be detected from each molecule. Particularly recording at cryogenic temperatures dramatically reduces photobleaching and would, hence, in principle, allow the user to massively increase the illumination time to several seconds. The downside of long illuminations, however, would be image blur due to inevitable jitter or drift occurring during the illuminations, which deteriorates the localization precision. In this paper, we theoretically demonstrate that a parallel recording of the fiducial marker beads together with a fitting approach accounting for the full drift trajectory allows for largely eliminating drift effects for drift magnitudes of several hundred nanometers per frame. We showcase the method for linear and diffusional drift as well as oscillations, assuming fixed dipole orientations during each illumination.

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07/10/23 | Input density tunes Kenyon cell sensory responses in the Drosophila mushroom body.
Ahmed M, Rajagopalan AE, Pan Y, Li Y, Williams DL, Pedersen EA, Thakral M, Previero A, Close KC, Christoforou CP, Cai D, Turner GC, Clowney EJ
Current Biology. 2023 Jul 10;33(13):2742-2760.e12. doi: 10.1016/j.cub.2023.05.064

The ability to discriminate sensory stimuli with overlapping features is thought to arise in brain structures called expansion layers, where neurons carrying information about sensory features make combinatorial connections onto a much larger set of cells. For 50 years, expansion coding has been a prime topic of theoretical neuroscience, which seeks to explain how quantitative parameters of the expansion circuit influence sensory sensitivity, discrimination, and generalization. Here, we investigate the developmental events that produce the quantitative parameters of the arthropod expansion layer, called the mushroom body. Using Drosophila melanogaster as a model, we employ genetic and chemical tools to engineer changes to circuit development. These allow us to produce living animals with hypothesis-driven variations on natural expansion layer wiring parameters. We then test the functional and behavioral consequences. By altering the number of expansion layer neurons (Kenyon cells) and their dendritic complexity, we find that input density, but not cell number, tunes neuronal odor selectivity. Simple odor discrimination behavior is maintained when the Kenyon cell number is reduced and augmented by Kenyon cell number expansion. Animals with increased input density to each Kenyon cell show increased overlap in Kenyon cell odor responses and become worse at odor discrimination tasks.

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07/07/23 | Volume electron microscopy.
Peddie CJ, Genoud C, Kreshuk A, Meechan K, Micheva KD, Narayan K, Pape C, Parton RG, Schieber NL, Schwab Y, Titze B, Verkade P, Aubrey A, Collinson LM
Nature Reviews: Methods Primers. 2022 Jul 07;2:51. doi: 10.1038/s43586-022-00131-9

Life exists in three dimensions, but until the turn of the century most electron microscopy methods provided only 2D image data. Recently, electron microscopy techniques capable of delving deep into the structure of cells and tissues have emerged, collectively called volume electron microscopy (vEM). Developments in vEM have been dubbed a quiet revolution as the field evolved from established transmission and scanning electron microscopy techniques, so early publications largely focused on the bioscience applications rather than the underlying technological breakthroughs. However, with an explosion in the uptake of vEM across the biosciences and fast-paced advances in volume, resolution, throughput and ease of use, it is timely to introduce the field to new audiences. In this Primer, we introduce the different vEM imaging modalities, the specialized sample processing and image analysis pipelines that accompany each modality and the types of information revealed in the data. We showcase key applications in the biosciences where vEM has helped make breakthrough discoveries and consider limitations and future directions. We aim to show new users how vEM can support discovery science in their own research fields and inspire broader uptake of the technology, finally allowing its full adoption into mainstream biological imaging.

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07/05/23 | A rise-to-threshold process for a relative-value decision.
Vijayan V, Wang F, Wang K, Chakravorty A, Adachi A, Akhlaghpour H, Dickson BJ, Maimon G
Nature. 2023 Jul 05;619(7970):563-571. doi: 10.1038/s41586-023-06271-6

Whereas progress has been made in the identification of neural signals related to rapid, cued decisions, less is known about how brains guide and terminate more ethologically relevant decisions in which an animal's own behaviour governs the options experienced over minutes. Drosophila search for many seconds to minutes for egg-laying sites with high relative value and have neurons, called oviDNs, whose activity fulfills necessity and sufficiency criteria for initiating the egg-deposition motor programme. Here we show that oviDNs express a calcium signal that (1) dips when an egg is internally prepared (ovulated), (2) drifts up and down over seconds to minutes-in a manner influenced by the relative value of substrates-as a fly determines whether to lay an egg and (3) reaches a consistent peak level just before the abdomen bend for egg deposition. This signal is apparent in the cell bodies of oviDNs in the brain and it probably reflects a behaviourally relevant rise-to-threshold process in the ventral nerve cord, where the synaptic terminals of oviDNs are located and where their output can influence behaviour. We provide perturbational evidence that the egg-deposition motor programme is initiated once this process hits a threshold and that subthreshold variation in this process regulates the time spent considering options and, ultimately, the choice taken. Finally, we identify a small recurrent circuit that feeds into oviDNs and show that activity in each of its constituent cell types is required for laying an egg. These results argue that a rise-to-threshold process regulates a relative-value, self-paced decision and provide initial insight into the underlying circuit mechanism for building this process.

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