60 results on '"Thuc Nghi Nguyen"'
Search Results
2. Single-cell and single-nucleus RNA-seq uncovers shared and distinct axes of variation in dorsal LGN neurons in mice, non-human primates, and humans
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Trygve E Bakken, Cindy TJ van Velthoven, Vilas Menon, Rebecca D Hodge, Zizhen Yao, Thuc Nghi Nguyen, Lucas T Graybuck, Gregory D Horwitz, Darren Bertagnolli, Jeff Goldy, Anna Marie Yanny, Emma Garren, Sheana Parry, Tamara Casper, Soraya I Shehata, Eliza R Barkan, Aaron Szafer, Boaz P Levi, Nick Dee, Kimberly A Smith, Susan M Sunkin, Amy Bernard, John Phillips, Michael J Hawrylycz, Christof Koch, Gabe J Murphy, Ed Lein, Hongkui Zeng, and Bosiljka Tasic
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maccaca nemestrina ,macaca fascicularis ,single-cell RNA-seq ,species comparison ,lateral geniculate nucleus ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Abundant evidence supports the presence of at least three distinct types of thalamocortical (TC) neurons in the primate dorsal lateral geniculate nucleus (dLGN) of the thalamus, the brain region that conveys visual information from the retina to the primary visual cortex (V1). Different types of TC neurons in mice, humans, and macaques have distinct morphologies, distinct connectivity patterns, and convey different aspects of visual information to the cortex. To investigate the molecular underpinnings of these cell types, and how these relate to differences in dLGN between human, macaque, and mice, we profiled gene expression in single nuclei and cells using RNA-sequencing. These efforts identified four distinct types of TC neurons in the primate dLGN: magnocellular (M) neurons, parvocellular (P) neurons, and two types of koniocellular (K) neurons. Despite extensively documented morphological and physiological differences between M and P neurons, we identified few genes with significant differential expression between transcriptomic cell types corresponding to these two neuronal populations. Likewise, the dominant feature of TC neurons of the adult mouse dLGN is high transcriptomic similarity, with an axis of heterogeneity that aligns with core vs. shell portions of mouse dLGN. Together, these data show that transcriptomic differences between principal cell types in the mature mammalian dLGN are subtle relative to the observed differences in morphology and cortical projection targets. Finally, alignment of transcriptome profiles across species highlights expanded diversity of GABAergic neurons in primate versus mouse dLGN and homologous types of TC neurons in primates that are distinct from TC neurons in mouse.
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- 2021
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3. Distinct Transcriptomic Cell Types and Neural Circuits of the Subiculum and Prosubiculum along the Dorsal-Ventral Axis
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Song-Lin Ding, Zizhen Yao, Karla E. Hirokawa, Thuc Nghi Nguyen, Lucas T. Graybuck, Olivia Fong, Phillip Bohn, Kiet Ngo, Kimberly A. Smith, Christof Koch, John W. Phillips, Ed S. Lein, Julie A. Harris, Bosiljka Tasic, and Hongkui Zeng
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scRNA-seq ,single-cell transcriptomics ,subicular complex ,prosubiculum ,ventral hippocampus ,cell types ,Biology (General) ,QH301-705.5 - Abstract
Summary: Subicular regions play important roles in spatial processing and many cognitive functions, and these are mainly attributed to the subiculum (Sub) rather than the prosubiculum (PS). Using single-cell RNA sequencing, we identify 27 transcriptomic cell types residing in sub-domains of the Sub and PS. Based on in situ expression of reliable transcriptomic markers, the precise boundaries of the Sub and PS are consistently defined along the dorsoventral axis. Using these borders to evaluate Cre-line specificity and tracer injections, we find bona fide Sub projections topographically to structures important for spatial processing and navigation. In contrast, the PS sends its outputs to widespread brain regions crucial for motivation, emotion, reward, stress, anxiety, and fear. The Sub and PS, respectively, dominate dorsal and ventral subicular regions and receive different afferents. These results reveal two molecularly and anatomically distinct circuits centered in the Sub and PS, respectively, providing a consistent explanation for historical data and a clearer foundation for future studies.
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- 2020
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4. Author Correction: Cell segmentation-free inference of cell types from in situ transcriptomics data
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Jeongbin Park, Wonyl Choi, Sebastian Tiesmeyer, Brian Long, Lars E. Borm, Emma Garren, Thuc Nghi Nguyen, Bosiljka Tasic, Simone Codeluppi, Tobias Graf, Matthias Schlesner, Oliver Stegle, Roland Eils, and Naveed Ishaque
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Science - Published
- 2021
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5. Single-nucleus and single-cell transcriptomes compared in matched cortical cell types.
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Trygve E Bakken, Rebecca D Hodge, Jeremy A Miller, Zizhen Yao, Thuc Nghi Nguyen, Brian Aevermann, Eliza Barkan, Darren Bertagnolli, Tamara Casper, Nick Dee, Emma Garren, Jeff Goldy, Lucas T Graybuck, Matthew Kroll, Roger S Lasken, Kanan Lathia, Sheana Parry, Christine Rimorin, Richard H Scheuermann, Nicholas J Schork, Soraya I Shehata, Michael Tieu, John W Phillips, Amy Bernard, Kimberly A Smith, Hongkui Zeng, Ed S Lein, and Bosiljka Tasic
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Medicine ,Science - Abstract
Transcriptomic profiling of complex tissues by single-nucleus RNA-sequencing (snRNA-seq) affords some advantages over single-cell RNA-sequencing (scRNA-seq). snRNA-seq provides less biased cellular coverage, does not appear to suffer cell isolation-based transcriptional artifacts, and can be applied to archived frozen specimens. We used well-matched snRNA-seq and scRNA-seq datasets from mouse visual cortex to compare cell type detection. Although more transcripts are detected in individual whole cells (~11,000 genes) than nuclei (~7,000 genes), we demonstrate that closely related neuronal cell types can be similarly discriminated with both methods if intronic sequences are included in snRNA-seq analysis. We estimate that the nuclear proportion of total cellular mRNA varies from 20% to over 50% for large and small pyramidal neurons, respectively. Together, these results illustrate the high information content of nuclear RNA for characterization of cellular diversity in brain tissues.
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- 2018
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6. Layer-specific chromatin accessibility landscapes reveal regulatory networks in adult mouse visual cortex
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Lucas T Gray, Zizhen Yao, Thuc Nghi Nguyen, Tae Kyung Kim, Hongkui Zeng, and Bosiljka Tasic
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visual cortex ,ATAC-seq ,transcription ,chromatin accessibility ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Mammalian cortex is a laminar structure, with each layer composed of a characteristic set of cell types with different morphological, electrophysiological, and connectional properties. Here, we define chromatin accessibility landscapes of major, layer-specific excitatory classes of neurons, and compare them to each other and to inhibitory cortical neurons using the Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq). We identify a large number of layer-specific accessible sites, and significant association with genes that are expressed in specific cortical layers. Integration of these data with layer-specific transcriptomic profiles and transcription factor binding motifs enabled us to construct a regulatory network revealing potential key layer-specific regulators, including Cux1/2, Foxp2, Nfia, Pou3f2, and Rorb. This dataset is a valuable resource for identifying candidate layer-specific cis-regulatory elements in adult mouse cortex.
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- 2017
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7. A high-resolution transcriptomic and spatial atlas of cell types in the whole mouse brain
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Zizhen Yao, Cindy T. J. van Velthoven, Michael Kunst, Meng Zhang, Delissa McMillen, Changkyu Lee, Won Jung, Jeff Goldy, Aliya Abdelhak, Pamela Baker, Eliza Barkan, Darren Bertagnolli, Jazmin Campos, Daniel Carey, Tamara Casper, Anish Bhaswanth Chakka, Rushil Chakrabarty, Sakshi Chavan, Min Chen, Michael Clark, Jennie Close, Kirsten Crichton, Scott Daniel, Tim Dolbeare, Lauren Ellingwood, James Gee, Alexandra Glandon, Jessica Gloe, Joshua Gould, James Gray, Nathan Guilford, Junitta Guzman, Daniel Hirschstein, Windy Ho, Kelly Jin, Matthew Kroll, Kanan Lathia, Arielle Leon, Brian Long, Zoe Maltzer, Naomi Martin, Rachel McCue, Emma Meyerdierks, Thuc Nghi Nguyen, Trangthanh Pham, Christine Rimorin, Augustin Ruiz, Nadiya Shapovalova, Cliff Slaughterbeck, Josef Sulc, Michael Tieu, Amy Torkelson, Herman Tung, Nasmil Valera Cuevas, Katherine Wadhwani, Katelyn Ward, Boaz Levi, Colin Farrell, Carol L. Thompson, Shoaib Mufti, Chelsea M. Pagan, Lauren Kruse, Nick Dee, Susan M. Sunkin, Luke Esposito, Michael J. Hawrylycz, Jack Waters, Lydia Ng, Kimberly A. Smith, Bosiljka Tasic, Xiaowei Zhuang, and Hongkui Zeng
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Article - Abstract
The mammalian brain is composed of millions to billions of cells that are organized into numerous cell types with specific spatial distribution patterns and structural and functional properties. An essential step towards understanding brain function is to obtain a parts list, i.e., a catalog of cell types, of the brain. Here, we report a comprehensive and high-resolution transcriptomic and spatial cell type atlas for the whole adult mouse brain. The cell type atlas was created based on the combination of two single-cell-level, whole-brain-scale datasets: a single- cell RNA-sequencing (scRNA-seq) dataset of ∼7 million cells profiled, and a spatially resolved transcriptomic dataset of ∼4.3 million cells using MERFISH. The atlas is hierarchically organized into five nested levels of classification: 7 divisions, 32 classes, 306 subclasses, 1,045 supertypes and 5,200 clusters. We systematically analyzed the neuronal, non-neuronal, and immature neuronal cell types across the brain and identified a high degree of correspondence between transcriptomic identity and spatial specificity for each cell type. The results reveal unique features of cell type organization in different brain regions, in particular, a dichotomy between the dorsal and ventral parts of the brain: the dorsal part contains relatively fewer yet highly divergent neuronal types, whereas the ventral part contains more numerous neuronal types that are more closely related to each other. We also systematically characterized cell-type specific expression of neurotransmitters, neuropeptides, and transcription factors. The study uncovered extraordinary diversity and heterogeneity in neurotransmitter and neuropeptide expression and co-expression patterns in different cell types across the brain, suggesting they mediate a myriad of modes of intercellular communications. Finally, we found that transcription factors are major determinants of cell type classification in the adult mouse brain and identified a combinatorial transcription factor code that defines cell types across all parts of the brain. The whole-mouse-brain transcriptomic and spatial cell type atlas establishes a benchmark reference atlas and a foundational resource for deep and integrative investigations of cell type and circuit function, development, and evolution of the mammalian brain.
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- 2023
8. Dense functional and molecular readout of a circuit hub in sensory cortex
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Condylis C, Bosiljka Tasic, Shenqin Yao, Thuc Nghi Nguyen, Chen Jl, Bistrong K, Manjrekar N, Hongkui Zeng, Zizhen Yao, and Abed Ghanbari
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Cell type ,Sensory processing ,medicine.medical_treatment ,Population ,Gene Expression ,Mice, Transgenic ,Sensory system ,Biology ,Somatosensory system ,Article ,Mice ,Calcium imaging ,medicine ,Animals ,Sensory cortex ,education ,Neurons ,education.field_of_study ,Neocortex ,Multidisciplinary ,Behavior, Animal ,Genes, fos ,Neural Inhibition ,Somatosensory Cortex ,Mice, Inbred C57BL ,Memory, Short-Term ,medicine.anatomical_structure ,Touch ,Vibrissae ,Calcium ,Nerve Net ,Transcriptome ,Neuroscience - Abstract
INTRODUCTION: The diversity of cell types is a defining feature of the neuronal circuitry that makes up the areas and layers of the mammalian cortex. At a molecular level, the extent of this diversity is now better appreciated through recent efforts to census all potential cortical cell types through single-cell transcriptional profiling. Cortical populations can be hierarchically subdivided into multiple putative transcriptomic cell classes, subclasses, and types. This new catalog of neuronal subclasses and subtypes opens up new questions and avenues of investigation for how these cell types are collectively organized into circuits that function to process information and adapt to changes in experience. RATIONALE: We investigated the function of newly identified cell types in layers 2 or 3 (L2/3) of the primary somatosensory cortex, a region that integrates bottom-up sensory information with top-down internal representations. Current in vivo methods primarily allow cell types to be investigated one at a time and have limited ability to label cell types defined by combinations of expressed genes. To densely survey these cell types and investigate how they interact during task behavior, we developed a platform, Comprehensive Readout of Activity and Cell Type Markers (CRACK), that combines population calcium imaging with subsequent multiplexed fluorescent in situ hybridization. Multiplexed labeling of mRNA transcripts is critical to deciphering the identity of cell types defined by combinatorial patterns of gene expression. RESULTS: We profiled the functional responses of three excitatory cell types and eight inhibitory subclasses in L2/3 as mice performed a whisker-based tactile working memory task. Task-related properties of both excitatory and inhibitory neurons continue to differentiate as they are segregated into increasingly discrete molecular types. Our analysis revealed that the excitatory cell type, L2/3 intratelencephalic Baz1a (Baz1a), functions as a highly active detector of tactile features. Simultaneous imaging across identified cell types enabled measurements of functional connectivity between subpopulations. Functional connectivity analysis indicated that Baz1a neurons orchestrate local network activity patterns. We found that Baz1a neurons show strong functional connections with dendrite-targeting, somatostatin-expressing (Sst) inhibitory neurons. Trans-monosynaptic viral tracing confirmed that Baz1a neurons preferentially synapse onto Sst neurons. Baz1a neurons also show enrichment of select plasticity-related, immediate early genes, including Fos. To determine whether the expression pattern of immediate early genes is a stable property of Baz1a neurons and how this relates to neuronal plasticity, we tracked Fos expression and neuronal activity in mice subjected to whisker deprivation. We found that Baz1a neurons homeostatically adapt to sensory deprivation while stably maintaining Fos expression. CONCLUSION: These results demonstrate that Baz1a neurons are a component of a molecularly defined circuit motif that is capable of recruiting local circuits for sensory processing when salient features are encountered during behavior. This cell type also functions to preserve sensory representations during ongoing and altered sensory experience. This builds on our knowledge for how local circuits in somatosensory cortex are implemented to negotiate bottom-up and top-down information. The ability to map functional and transcriptional relationships across neuronal populations provides insight into how the organizing principles of the cortex give rise to the computations it performs.
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- 2022
9. Author Correction: Cell segmentation-free inference of cell types from in situ transcriptomics data
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Naveed Ishaque, Tobias Graf, Lars E. Borm, Sebastian Tiesmeyer, Matthias Schlesner, Bosiljka Tasic, Oliver Stegle, Roland Eils, Wonyl Choi, Simone Codeluppi, Emma Garren, Brian Long, Thuc Nghi Nguyen, and Jeongbin Park
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In situ ,Transcriptome ,Cell type ,Multidisciplinary ,Computer science ,Science ,General Physics and Astronomy ,Cell segmentation ,Inference ,General Chemistry ,Computational biology ,General Biochemistry, Genetics and Molecular Biology - Published
- 2021
10. Author response: Single-cell and single-nucleus RNA-seq uncovers shared and distinct axes of variation in dorsal LGN neurons in mice, non-human primates, and humans
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Amy Bernard, Aaron Szafer, Nick Dee, Michael Hawrylycz, Susan M. Sunkin, Ed S. Lein, Rebecca D. Hodge, Soraya I. Shehata, John W. Phillips, Gregory D. Horwitz, Emma Garren, Jeff Goldy, Christof Koch, Eliza Barkan, Zizhen Yao, Thuc Nghi Nguyen, Kimberly A. Smith, Sheana Parry, Lucas T. Graybuck, Anna Marie Yanny, Tamara Casper, Darren Bertagnolli, Hongkui Zeng, Bosiljka Tasic, Cindy T. J. van Velthoven, Boaz P. Levi, Trygve E. Bakken, Vilas Menon, and Gabe J. Murphy
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Dorsum ,Variation (linguistics) ,medicine.anatomical_structure ,Cell ,medicine ,RNA-Seq ,Biology ,Nucleus ,Cell biology - Published
- 2021
11. Characterization of a fiber-coupled EvenField illumination system for fluorescence microscopy
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Kristen Turner, Kyla Berry, Robert Serafin, Mike Taormina, Melissa Gorham, Zoe Maltzer, Thuc Nghi Nguyen, and Philip R Nicovich
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Multi-mode optical fiber ,Materials science ,Speckle reduction ,business.industry ,ComputingMethodologies_IMAGEPROCESSINGANDCOMPUTERVISION ,Speckle noise ,Atomic and Molecular Physics, and Optics ,Characterization (materials science) ,Reduction (complexity) ,Speckle pattern ,Optics ,Homogeneous ,Fluorescence microscope ,Image sensor ,A fibers ,business - Abstract
Fluorescence microscopy benefits from spatially and temporally homogeneous illumination with illumination area matched to the shape and size of the camera sensor. Fiber-coupled illumination schemes have the added benefit of straightforward and robust alignment and ease of installation compared to free-space coupled illumination. Commercial and open-source fiber-coupled, homogenized illumination schemes have recently become available to the public; however, there have been no published comparisons of speckle reduction schemes to date. We characterize three different multimode fibers in combination with two laser speckle reduction devices and compare spatial and temporal profiles to a commercial unit. This work yields a new design, the EvenField Illuminator, which is freely available along for researchers to integrate into their own imaging systems.
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- 2021
12. Classification of electrophysiological and morphological neuron types in the mouse visual cortex
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David Sandman, Brian Lee, Michael Hawrylycz, Sara Kebede, Tom Egdorf, David Reid, Rob Young, Nivretta Thatra, Stefan Mihalas, David Feng, John W. Phillips, Rebecca de Frates, DiJon Hill, Cliff Slaughterbeck, Samuel R Josephsen, Tamara Casper, Xiaoxiao Liu, Hanchuan Peng, Peter Chong, Colin Farrell, Zhi Zhou, Sheana Parry, Jed Perkins, Brian Long, Susan M. Sunkin, Matthew Kroll, Krissy Brouner, Melissa Gorham, Aaron Szafer, Wayne Wakeman, Hong Gu, Marissa Garwood, Daniel Park, Kristen Hadley, Michael S. Fisher, Lydia Potekhina, Ed Lein, Alice Mukora, Hongkui Zeng, Nick Dee, Aaron Oldre, Lindsay Ng, Thomas Braun, Grace Williams, Tracy Lemon, Julie A. Harris, Medea McGraw, Nadezhda Dotson, Philip R. Nicovich, Amanda Gary, Rusty Mann, Alex M. Henry, Caroline Habel, Samuel Dingman, Katherine E. Link, Nathalie Gaudreault, Gilberto J. Soler-Llavina, Thuc Nghi Nguyen, Nicole Blesie, Bosiljka Tasic, Lydia Ng, Christine Cuhaciyan, Tim Jarsky, Keith B. Godfrey, Costas A. Anastassiou, Kirsten Crichton, Josef Sulc, Martin Schroedter, Dan Castelli, Miranda Robertson, Amy Bernard, Lisa Kim, Songlin Ding, Alyse Doperalski, Nathan W. Gouwens, Herman Tung, Tsega Desta, Corinne Teeter, James Harrington, Jonathan T. Ting, Kris Bickley, Anton Arkhipov, Kiet Ngo, Changkyu Lee, Jim Berg, Agata Budzillo, Emma Garren, Tanya L. Daigle, Christof Koch, Rachel A. Dalley, Eliza Barkan, Staci A. Sorensen, Gabe J. Murphy, Shiella Caldejon, and Naz Taskin
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0301 basic medicine ,Genetically modified mouse ,Cell type ,Patch-Clamp Techniques ,Databases, Factual ,Action Potentials ,Datasets as Topic ,Mice, Transgenic ,Biology ,Article ,Neuron types ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Genes, Reporter ,Biocytin ,medicine ,Animals ,Cell shape ,Cell Shape ,Visual Cortex ,Neurons ,General Neuroscience ,Laboratory mouse ,Electrophysiology ,030104 developmental biology ,Visual cortex ,medicine.anatomical_structure ,chemistry ,Transcriptome ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Understanding the diversity of cell types in the brain has been an enduring challenge and requires detailed characterization of individual neurons in multiple dimensions. To systematically profile morpho-electric properties of mammalian neurons, we established a single-cell characterization pipeline using standardized patch-clamp recordings in brain slices and biocytin-based neuronal reconstructions. We built a publicly accessible online database, the Allen Cell Types Database, to display these datasets. Intrinsic physiological properties were measured from 1,938 neurons from the adult laboratory mouse visual cortex, morphological properties were measured from 461 reconstructed neurons, and 452 neurons had both measurements available. Quantitative features were used to classify neurons into distinct types using unsupervised methods. We established a taxonomy of morphologically and electrophysiologically defined cell types for this region of the cortex, with 17 electrophysiological types, 38 morphological types and 46 morpho-electric types. There was good correspondence with previously defined transcriptomic cell types and subclasses using the same transgenic mouse lines.
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- 2019
13. Distinct descending motor cortex pathways and their roles in movement
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Vilas Menon, Johan Winnubst, Lucas T. Graybuck, Hongkui Zeng, Charles R. Gerfen, Thuc Nghi Nguyen, Zizhen Yao, Lihua Wang, Bosiljka Tasic, Erhan Bas, Karel Svoboda, Loren L. Looger, Jayaram Chandrashekar, Kimberly A. Smith, Sarada Viswanathan, and Michael N. Economo
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0301 basic medicine ,Multidisciplinary ,Pyramidal tracts ,Motor control ,Biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,Basal ganglia ,medicine ,Biological neural network ,Neuron ,Brainstem ,Neuroscience ,030217 neurology & neurosurgery ,Medulla ,Motor cortex - Abstract
Activity in the motor cortex predicts movements, seconds before they are initiated. This preparatory activity has been observed across cortical layers, including in descending pyramidal tract neurons in layer 5. A key question is how preparatory activity is maintained without causing movement, and is ultimately converted to a motor command to trigger appropriate movements. Here, using single-cell transcriptional profiling and axonal reconstructions, we identify two types of pyramidal tract neuron. Both types project to several targets in the basal ganglia and brainstem. One type projects to thalamic regions that connect back to motor cortex; populations of these neurons produced early preparatory activity that persisted until the movement was initiated. The second type projects to motor centres in the medulla and mainly produced late preparatory activity and motor commands. These results indicate that two types of motor cortex output neurons have specialized roles in motor control.
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- 2018
14. Single-cell and single-nucleus RNA-seq uncovers shared and distinct axes of variation in dorsal LGN neurons in mice, non-human primates, and humans
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Ed S. Lein, Vilas Menon, John W. Phillips, Sheana Parry, Jeff Goldy, Lucas T. Graybuck, Cindy T. J. van Velthoven, Michael Hawrylycz, Kimberly A. Smith, Susan M. Sunkin, Amy Bernard, Christof Koch, Zizhen Yao, Aaron Szafer, Nick Dee, Bosiljka Tasic, Anna Marie Yanny, Rebecca D. Hodge, Hongkui Zeng, Darren Bertagnolli, Tamara Casper, Boaz P. Levi, Trygve E. Bakken, Thuc Nghi Nguyen, Gabe J. Murphy, Eliza Barkan, Emma Garren, Soraya I. Shehata, and Gregory D. Horwitz
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Cell type ,Mouse ,QH301-705.5 ,species comparison ,Science ,macaca fascicularis ,Thalamus ,Lateral geniculate nucleus ,Macaque ,General Biochemistry, Genetics and Molecular Biology ,Mice ,lateral geniculate nucleus ,Parvocellular cell ,biology.animal ,medicine ,Animals ,Humans ,Visual Pathways ,RNA-Seq ,Biology (General) ,Visual Cortex ,Cell Nucleus ,Neurons ,single-cell RNA-seq ,General Immunology and Microbiology ,biology ,General Neuroscience ,Gene Expression Profiling ,Geniculate Bodies ,General Medicine ,Koniocellular cell ,Visual cortex ,medicine.anatomical_structure ,nervous system ,maccaca nemestrina ,Medicine ,Macaca ,Other ,Single-Cell Analysis ,Neuroscience ,Nucleus ,Research Article ,Human - Abstract
Abundant evidence supports the presence of at least three distinct types of thalamocortical (TC) neurons in the primate dorsal lateral geniculate nucleus (dLGN) of the thalamus, the brain region that conveys visual information from the retina to the primary visual cortex (V1). Different types of TC neurons in mice, humans, and macaques have distinct morphologies, distinct connectivity patterns, and convey different aspects of visual information to the cortex. To investigate the molecular underpinnings of these cell types, and how these relate to differences in dLGN between human, macaque, and mice, we profiled gene expression in single nuclei and cells using RNA-sequencing. These efforts identified four distinct types of TC neurons in the primate dLGN: magnocellular (M) neurons, parvocellular (P) neurons, and two types of koniocellular (K) neurons. Despite extensively documented morphological and physiological differences between M and P neurons, we identified few genes with significant differential expression between transcriptomic cell types corresponding to these two neuronal populations. Likewise, the dominant feature of TC neurons of the adult mouse dLGN is high transcriptomic similarity, with an axis of heterogeneity that aligns with core vs. shell portions of mouse dLGN. Together, these data show that transcriptomic differences between principal cell types in the mature mammalian dLGN are subtle relative to the observed differences in morphology and cortical projection targets. Finally, alignment of transcriptome profiles across species highlights expanded diversity of GABAergic neurons in primate versus mouse dLGN and homologous types of TC neurons in primates that are distinct from TC neurons in mouse.
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- 2020
15. Brain-wide single neuron reconstruction reveals morphological diversity in molecularly defined striatal, thalamic, cortical and claustral neuron types
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Susan M. Sunkin, Zizhen Yao, Qi Li, Tanya L. Daigle, Yun Wang, Michael Hawrylycz, Jia Yuan, Donghuan Lu, Bosiljka Tasic, Lulu Yin, Yuanyuan Song, Z. Josh Huang, Karla E. Hirokawa, Zheng Yefeng, Matthew B. Veldman, Lei Huang, Luke Esposito, Feng Xiong, Shaoqun Zeng, An Liu, Liya Ding, Guodong Hong, Jintao Pan, Yaoyao Li, Wei Xiong, Qiang Ouyang, Yang Yu, Thuc Nghi Nguyen, Qingming Luo, Yimin Wang, Xiangning Li, Mengya Chen, Tao Wang, Zhangcan Ding, Lei Qu, Lydia Ng, Min Ye, Hsien-Chi Kuo, Peng Xie, Yuanyuan Li, Rachael Larsen, Zhixi Yun, Chris Hill, Julie A. Harris, Peng Wang, Longfei Li, Elise Shen, Lijuan Liu, Wan Wan, Sujun Zhao, Hui Gong, Zhongze Gu, Zongcai Ruan, Jing Yuan, Christof Koch, Xiangdong Yang, Wenjie Xu, Hongkui Zeng, Aaron Feiner, Stephanie Mok, Yanjun Duan, Shichen Zhang, Chao Chen, Yaping Wang, Wayne Wakeman, Phil Lesnar, Sara Kebede, Ping He, Staci A. Sorensen, Zijun Zhao, Anan Li, Hanchuan Peng, Xiuli Kuang, Shengdian Jiang, Zhi Zhou, Quanxin Wang, and Wei Xie
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Cell type ,medicine.anatomical_structure ,nervous system ,Cortex (anatomy) ,Thalamus ,medicine ,Striatum ,Neuron ,Biology ,Axon ,Projection (set theory) ,Claustrum ,Neuroscience - Abstract
Ever since the seminal findings of Ramon y Cajal, dendritic and axonal morphology has been recognized as a defining feature of neuronal types. Yet our knowledge concerning the diversity of neuronal morphologies, in particular distal axonal projection patterns, is extremely limited. To systematically obtain single neuron full morphology on a brain-wide scale, we established a platform with five major components: sparse labeling, whole-brain imaging, reconstruction, registration, and classification. We achieved sparse, robust and consistent fluorescent labeling of a wide range of neuronal types by combining transgenic or viral Cre delivery with novel transgenic reporter lines. We acquired high-resolution whole-brain fluorescent images from a large set of sparsely labeled brains using fluorescence micro-optical sectioning tomography (fMOST). We developed a set of software tools for efficient large-volume image data processing, registration to the Allen Mouse Brain Common Coordinate Framework (CCF), and computer-assisted morphological reconstruction. We reconstructed and analyzed the complete morphologies of 1,708 neurons from the striatum, thalamus, cortex and claustrum. Finally, we classified these cells into multiple morphological and projection types and identified a set of region-specific organizational rules of long-range axonal projections at the single cell level. Specifically, different neuron types from different regions follow highly distinct rules in convergent or divergent projection, feedforward or feedback axon termination patterns, and between-cell homogeneity or heterogeneity. Major molecularly defined classes or types of neurons have correspondingly distinct morphological and projection patterns, however, we also identify further remarkably extensive morphological and projection diversity at more fine-grained levels within the major types that cannot presently be accounted for by preexisting transcriptomic subtypes. These insights reinforce the importance of full morphological characterization of brain cell types and suggest a plethora of ways different cell types and individual neurons may contribute to the function of their respective circuits.
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- 2020
16. Morphological diversity of single neurons in molecularly defined cell types
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Anan Li, Elise Shen, Lijuan Liu, Wan Wan, Hui Gong, Yanjun Duan, Rachel A. Dalley, Sujun Zhao, Luke Esposito, Zhixi Yun, Shaoqun Zeng, An Liu, Susan M. Sunkin, Zhi Zhou, Tanya L. Daigle, Jintao Pan, Liya Ding, Yaoyao Li, Chris Hill, Yimin Wang, Yefeng Zheng, Qingming Luo, Phil Lesnar, Karla E. Hirokawa, Zijun Zhao, Christof Koch, Qi Li, Ping He, Donghuan Lu, Staci A. Sorensen, Longfei Li, Zhongze Gu, Xiangning Li, Zhangcan Ding, Lei Qu, Jia Yuan, Hsien-Chi Kuo, Aaron Feiner, Stephanie Mok, Julie A. Harris, Jing Yuan, Yang Yu, Qiang Ouyang, Z. Josh Huang, X. William Yang, Guodong Hong, Thuc Nghi Nguyen, Rachael Larsen, Michael Hawrylycz, Wenjie Xu, Peng Wang, Chao Chen, Wei Xiong, Hongkui Zeng, Mengya Chen, Zongcai Ruan, Feng Xiong, Shichen Zhang, Lydia Ng, Min Ye, Wayne Wakeman, Peng Xie, Yaping Wang, Quanxin Wang, Yun Wang, Sara Kebede, Bosiljka Tasic, Lulu Yin, Yuanyuan Song, Tao Wang, Lei Huang, Wei Xie, Zizhen Yao, Matthew B. Veldman, Yuanyuan Li, Xiuli Kuang, Shengdian Jiang, and Hanchuan Peng
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Cell type ,Neurogenesis ,Neocortex ,Biology ,Neural circuits ,Article ,Atlases as Topic ,Cellular neuroscience ,Biological neural network ,medicine ,Feature (machine learning) ,Humans ,RNA-Seq ,Axon ,Projection (set theory) ,Cell Shape ,Neurons ,Multidisciplinary ,Brain ,Gene Expression Regulation, Developmental ,Reproducibility of Results ,Claustrum ,medicine.anatomical_structure ,Evolutionary biology ,Single-Cell Analysis ,Neuroglia ,Function (biology) ,Biomarkers - Abstract
Dendritic and axonal morphology reflects the input and output of neurons and is a defining feature of neuronal types1,2, yet our knowledge of its diversity remains limited. Here, to systematically examine complete single-neuron morphologies on a brain-wide scale, we established a pipeline encompassing sparse labelling, whole-brain imaging, reconstruction, registration and analysis. We fully reconstructed 1,741 neurons from cortex, claustrum, thalamus, striatum and other brain regions in mice. We identified 11 major projection neuron types with distinct morphological features and corresponding transcriptomic identities. Extensive projectional diversity was found within each of these major types, on the basis of which some types were clustered into more refined subtypes. This diversity follows a set of generalizable principles that govern long-range axonal projections at different levels, including molecular correspondence, divergent or convergent projection, axon termination pattern, regional specificity, topography, and individual cell variability. Although clear concordance with transcriptomic profiles is evident at the level of major projection type, fine-grained morphological diversity often does not readily correlate with transcriptomic subtypes derived from unsupervised clustering, highlighting the need for single-cell cross-modality studies. Overall, our study demonstrates the crucial need for quantitative description of complete single-cell anatomy in cell-type classification, as single-cell morphological diversity reveals a plethora of ways in which different cell types and their individual members may contribute to the configuration and function of their respective circuits., Sparse labelling and whole-brain imaging are used to reconstruct and classify brain-wide complete morphologies of 1,741 individual neurons in the mouse brain, revealing a dependence on both brain region and transcriptomic profile.
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- 2020
17. Regional, layer, and cell-class specific connectivity of the mouse default mode network
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Ali Williford, Nile Graddis, Karla E. Hirokawa, Wayne Wakeman, Stefan Mihalas, Philip R. Nicovich, Thuc Nghi Nguyen, Olivia Fong, Adam Liska, Phillip Bohn, Anh Ho, Lydia Ng, Emma Garren, Boaz P. Levi, Kimberly A. Smith, Nick Dee, Julie A. Harris, David Feng, Alex M. Henry, Cindy T. J. van Velthoven, Peter A. Groblewski, Alessandro Gozzi, Jennifer D. Whitesell, Hongkui Zeng, Bosiljka Tasic, Maitham Naeemi, Joseph E. Knox, Leonard Kuan, and Ludovico Coletta
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Resting state functional magnetic resonance imaging ,Cell type ,Then test ,medicine.anatomical_structure ,Retrosplenial cortex ,Cell ,medicine ,Neuron ,Biology ,Neuroscience ,Multiple disorders ,human activities ,Default mode network - Abstract
The evolutionarily conserved default mode network (DMN) is characterized by temporally correlated activity between brain regions during resting states. The DMN has emerged as a selectively vulnerable network in multiple disorders, so understanding its anatomical composition will provide fundamental insight into how its function is impacted by disease. Reproducible rodent analogs of the human DMN offer an opportunity to investigate the underlying brain regions and structural connectivity (SC) with high spatial and cell type resolution. Here, we performed systematic analyses using mouse resting state functional magnetic resonance imaging to identify the DMN and whole brain axonal tracing data, co-registered to the 3D Allen Mouse Common Coordinate Framework reference atlas. We identified the specific, predominantly cortical, brain regions comprising the mouse DMN and report preferential SC between these regions. Next, at the cell class level, we report that cortical layer (L) 2/3 neurons in DMN regions project almost exclusively to other DMN regions, whereas L5 neurons project to targets both in and out of the DMN. We then test the hypothesis that in- and out-DMN projection patterns originate from distinct L5 neuron sub-classes using an intersectional viral tracing strategy to label all the axons from neurons defined by a single target. In the ventral retrosplenial cortex, a core DMN region, we found two L5 projection types related to the DMN and mapped them to unique transcriptomically-defined cell types. Together, our results provide a multi-scale description of the anatomical correlates of the mouse DMN.
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- 2020
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18. A taxonomy of transcriptomic cell types across the isocortex and hippocampal formation
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Susan M. Sunkin, Qingzhong Ren, Michael Tieu, Fahimeh Baftizadeh, Kimberly A. Smith, Boaz P. Levi, Kanan Lathia, Olivia Fong, James Gray, Lucas T. Graybuck, Jeff Goldy, Bosiljka Tasic, Christine Rimorin, Thuc Nghi Nguyen, Kirsten Crichton, Josef Sulc, Songlin Ding, Darren Bertagnolli, Zizhen Yao, Hongkui Zeng, Delissa McMillen, Cindy T. J. van Velthoven, Katelyn Ward, Alexandra Glandon, Thanh Pham, Herman Tung, Amy Torkelson, Nick Dee, Nadiya V. Shapovalova, Stephanie Mok, Emma Garren, Matthew Kroll, Tamara Casper, Adriana E. Sedeno-Cortes, and Daniel Hirschstein
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Transcriptome ,Glutamatergic ,Cell type ,Cellular composition ,Spatial distribution pattern ,Biology ,Hippocampal formation ,GABAergic neuron ,Neuroscience ,Neuron types - Abstract
SUMMARYThe isocortex and hippocampal formation are two major structures in the mammalian brain that play critical roles in perception, cognition, emotion and learning. Both structures contain multiple regions, for many of which the cellular composition is still poorly understood. In this study, we used two complementary single-cell RNA-sequencing approaches, SMART-Seq and 10x, to profile ∼1.2 million cells covering all regions in the adult mouse isocortex and hippocampal formation, and derived a cell type taxonomy comprising 379 transcriptomic types. The completeness of coverage enabled us to define gene expression variations across the entire spatial landscape without significant gaps. We found that cell types are organized in a hierarchical manner and exhibit varying degrees of discrete or continuous relatedness with each other. Such molecular relationships correlate strongly with the spatial distribution patterns of the cell types, which can be region-specific, or shared across multiple regions, or part of one or more gradients along with other cell types. Glutamatergic neuron types have much greater diversity than GABAergic neuron types, both molecularly and spatially, and they define regional identities as well as inter-region relationships. For example, we found that glutamatergic cell types between the isocortex and hippocampal formation are highly distinct from each other yet possess shared molecular signatures and corresponding layer specificities, indicating their homologous relationships. Overall, our study establishes a molecular architecture of the mammalian isocortex and hippocampal formation for the first time, and begins to shed light on its underlying relationship with the development, evolution, connectivity and function of these two brain structures.
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- 2020
19. A transcriptomic and epigenomic cell atlas of the mouse primary motor cortex
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Cindy T. J. van Velthoven, Eran A. Mukamel, Kanan Lathia, Jeff Goldy, Elizabeth Purdom, Hanqing Liu, Zizhen Yao, Xinxin Wang, Victor Felix, Olivia Fong, Brian R. Herb, Jacinta Lucero, Elizabeth L. Dougherty, Carlo Colantuoni, Thanh Pham, Bing Ren, Valentine Svensson, Sheng-Yong Niu, Rongxin Fang, Davide Risso, Seth A. Ament, Michael Tieu, Christine Rimorin, John Ngai, Ricky S. Adkins, Sandrine Dudoit, Naeem Nadaf, Stephan Fischer, Michael Hawrylycz, Evan Z. Macosko, Anup Mahurkar, Joshua Orvis, Lior Pachter, Thuc Nghi Nguyen, Peter V. Kharchenko, Vasilis Ntranos, Bosiljka Tasic, Joshua D. Welch, Darren Bertagnolli, Charles R. Vanderburg, Yang Eric Li, Eeshit Dhaval Vaishnav, Xiaomeng Hou, Joseph R. Ecker, Delissa McMillen, Kirsten Crichton, Heather Huot Creasy, Antonio Pinto-Duarte, Josef Sulc, A. Sina Booeshaghi, Megan Crow, Chongyuan Luo, Owen White, Kimberly A. Smith, Jayaram Kancherla, Jonathan Crabtree, Herman Tung, Wayne I. Doyle, Angeline Rivkin, M. Margarita Behrens, Hongkui Zeng, Kelly Street, Amy Torkelson, Tommaso Biancalani, Julia K. Osteen, Héctor Corrada Bravo, Aviv Regev, Anna Bartlett, Olivier Poirion, Nick Dee, Qiwen Hu, Michelle G. Giglio, Z. Josh Huang, Andrew Aldridge, Ronna Hertzano, Sebastian Preissl, Matthew Kroll, Koen Van den Berge, Fangming Xie, Jesse Gillis, Joseph R. Nery, Tamara Casper, and Hector Roux de Bézieux
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Epigenomics ,Male ,Cell type ,General Science & Technology ,1.1 Normal biological development and functioning ,Population ,Datasets as Topic ,Bioengineering ,Molecular neuroscience ,Computational biology ,Biology ,CLASSIFICATION ,Article ,Epigenesis, Genetic ,Mice ,Atlases as Topic ,Single-cell analysis ,Genetic ,Underpinning research ,MAPS ,medicine ,Genetics ,Animals ,education ,Neurons ,education.field_of_study ,ARCHITECTURE ,Multidisciplinary ,Gene Expression Profiling ,Human Genome ,Neurosciences ,Motor Cortex ,Biology and Life Sciences ,Reproducibility of Results ,Cellular neuroscience ,Gene expression profiling ,medicine.anatomical_structure ,Mathematics and Statistics ,Organ Specificity ,Neurological ,Female ,Primary motor cortex ,Single-Cell Analysis ,Transcriptome ,Motor cortex ,Epigenesis ,Biotechnology - Abstract
Single-cell transcriptomics can provide quantitative molecular signatures for large, unbiased samples of the diverse cell types in the brain1–3. With the proliferation of multi-omics datasets, a major challenge is to validate and integrate results into a biological understanding of cell-type organization. Here we generated transcriptomes and epigenomes from more than 500,000 individual cells in the mouse primary motor cortex, a structure that has an evolutionarily conserved role in locomotion. We developed computational and statistical methods to integrate multimodal data and quantitatively validate cell-type reproducibility. The resulting reference atlas—containing over 56 neuronal cell types that are highly replicable across analysis methods, sequencing technologies and modalities—is a comprehensive molecular and genomic account of the diverse neuronal and non-neuronal cell types in the mouse primary motor cortex. The atlas includes a population of excitatory neurons that resemble pyramidal cells in layer 4 in other cortical regions4. We further discovered thousands of concordant marker genes and gene regulatory elements for these cell types. Our results highlight the complex molecular regulation of cell types in the brain and will directly enable the design of reagents to target specific cell types in the mouse primary motor cortex for functional analysis., The authors describe an integrated atlas of the diverse cell types in the mouse primary motor cortex.
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- 2020
20. An integrated transcriptomic and epigenomic atlas of mouse primary motor cortex cell types
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Z. Josh Huang, Valentine Svensson, Christine Rimorin, Sebastian Preissl, Qiwen Hu, Yang Eric Li, Carlo Colantuoni, Olivier Poirion, Darren Bertagnolli, Vasilis Ntranos, Antonio Pinto-Duarte, Megan Crow, Delissa McMillen, Evan Z. Macosko, Nick Dee, Zizhen Yao, Hongkui Zeng, Hector Roux de Bézieux, Bing Ren, Sheng-Yong Niu, Brian R. Herb, Jacinta Lucero, Ricky S. Adkins, Rongxin Fang, Eeshit Dhaval Vaishnav, Peter V. Kharchenko, Charles R. Vanderburg, Xiaomeng Hou, Joshua D. Welch, Angeline Rivkin, Sandrine Dudoit, Michael Tieu, Michael Hawrylycz, Jayaram Kancherla, Anup Mahurkar, Victor Felix, Lior Pachter, Jonathan Crabtree, Ronna Hertzano, Héctor Corrada Bravo, Aviv Regev, Wayne I. Doyle, Fangming Xie, Owen White, A. Sina Booeshaghi, Chongyuan Luo, Jeff Goldy, Andrew I. Aldrige, Joseph R. Ecker, Naeem Nadaf, Elizabeth Purdom, Hanqing Liu, Eran A. Mukamel, Kanan Lathia, Kelly Street, Michelle G. Giglio, Xinxin Wang, Julia K. Osteen, Olivia Fong, Bosiljka Tasic, Matthew Kroll, Tommaso Biancalani, Thanh Pham, John Ngai, Amy Torkelson, Thuc Nghi Nguyen, Ann Bartlett, Kimberly A. Smith, Kirsten Crichton, Herman Tung, Heather Huot Creasy, Josef Sulc, M. Margarita Behrens, Cindy T. J. van Velthoven, Koen Van den Berge, Jesse Gillis, Joseph R. Nery, Tamara Casper, Elizabeth L. Dougherty, Davide Risso, Seth A. Ament, Stephan Fischer, and Joshua Orvis
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0303 health sciences ,Cell type ,Cell ,Computational biology ,Epigenome ,Biology ,Transcriptome ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,medicine ,Primary motor cortex ,Nucleus ,Gene ,030217 neurology & neurosurgery ,030304 developmental biology ,Epigenomics - Abstract
Single cell transcriptomics has transformed the characterization of brain cell identity by providing quantitative molecular signatures for large, unbiased samples of brain cell populations. With the proliferation of taxonomies based on individual datasets, a major challenge is to integrate and validate results toward defining biologically meaningful cell types. We used a battery of single-cell transcriptome and epigenome measurements generated by the BRAIN Initiative Cell Census Network (BICCN) to comprehensively assess the molecular signatures of cell types in the mouse primary motor cortex (MOp). We further developed computational and statistical methods to integrate these multimodal data and quantitatively validate the reproducibility of the cell types. The reference atlas, based on more than 600,000 high quality single-cell or -nucleus samples assayed by six molecular modalities, is a comprehensive molecular account of the diverse neuronal and non-neuronal cell types in MOp. Collectively, our study indicates that the mouse primary motor cortex contains over 55 neuronal cell types that are highly replicable across analysis methods, sequencing technologies, and modalities. We find many concordant multimodal markers for each cell type, as well as thousands of genes and gene regulatory elements with discrepant transcriptomic and epigenomic signatures. These data highlight the complex molecular regulation of brain cell types and will directly enable design of reagents to target specific MOp cell types for functional analysis.
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- 2020
21. Regional, Layer, and Cell-Type-Specific Connectivity of the Mouse Default Mode Network
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Phillip Bohn, Lydia Ng, Maitham Naeemi, Thuc Nghi Nguyen, Karla E. Hirokawa, Stefan Mihalas, Ali Williford, Kimberly A. Smith, Leonard Kuan, Joseph E. Knox, Nick Dee, Hongkui Zeng, Julie A. Harris, Ludovico Coletta, Alex M. Henry, Peter A. Groblewski, Olivia Fong, Adam Liska, Nile Graddis, Anh Ho, David Feng, Cindy T. J. van Velthoven, Wayne Wakeman, Jennifer D. Whitesell, Bosiljka Tasic, Boaz P. Levi, Alessandro Gozzi, Philip R. Nicovich, and Emma Garren
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0301 basic medicine ,retrosplenial cortex ,Single cell transcriptomics ,Cell type specific ,Population ,Biology ,single cell transcriptomics ,Axonal tracing ,Article ,projection neuron types ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Retrosplenial cortex ,Connectome ,medicine ,Animals ,DMN ,Layer (object-oriented design) ,education ,Default mode network ,axonal projections ,Neurons ,education.field_of_study ,medicine.diagnostic_test ,General Neuroscience ,Brain ,Default Mode Network ,Magnetic Resonance Imaging ,030104 developmental biology ,connectivity ,cortical connectome ,Nerve Net ,Functional magnetic resonance imaging ,viral tracer ,human activities ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Summary The evolutionarily conserved default mode network (DMN) is a distributed set of brain regions coactivated during resting states that is vulnerable to brain disorders. How disease affects the DMN is unknown, but detailed anatomical descriptions could provide clues. Mice offer an opportunity to investigate structural connectivity of the DMN across spatial scales with cell-type resolution. We co-registered maps from functional magnetic resonance imaging and axonal tracing experiments into the 3D Allen mouse brain reference atlas. We find that the mouse DMN consists of preferentially interconnected cortical regions. As a population, DMN layer 2/3 (L2/3) neurons project almost exclusively to other DMN regions, whereas L5 neurons project in and out of the DMN. In the retrosplenial cortex, a core DMN region, we identify two L5 projection types differentiated by in- or out-DMN targets, laminar position, and gene expression. These results provide a multi-scale description of the anatomical correlates of the mouse DMN., Graphical Abstract, Highlights • Mouse resting-state default mode network anatomy described at high resolution in 3D • Systematic axon tracing shows cortical DMN regions are preferentially interconnected • Layer 2/3 DMN neurons project mostly in the DMN; layer 5 neurons project in and out • Retrosplenial cortex contains distinct types of in- and out-DMN projection neurons, The default mode network is vulnerable to brain disorders, but details of its anatomy and connectivity are coarse. Whitesell et al. use modern neuroanatomical tools in the mouse, including whole-brain imaging and viral tracing, to provide high-resolution anatomical descriptions and identify cell type correlates of this conserved brain network.
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- 2020
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22. A Taxonomy of Transcriptomic Cell Types Across the Isocortex and Hippocampal Formation
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James Gray, Adriana E. Sedeno-Cortes, Michael Tieu, Songlin Ding, Michael Hawrylycz, Herman Tung, Olivia Fong, Matthew Kroll, Stephanie Mok, Zizhen Yao, Darren Bertagnolli, Fahimeh Baftizadeh, Thanh Pham, Delissa McMillen, Thuc Nghi Nguyen, Hongkui Zeng, Tamara Casper, Katelyn Ward, Emma Garren, Kimberly A. Smith, Qingzhong Ren, Christine Rimorin, Jeff Goldy, Alexandra Glandon, Kanan Lathia, Lucas T. Graybuck, Amy Torkelson, Nick Dee, Nadiya V. Shapovalova, Susan M. Sunkin, Daniel Hirschstein, Bosiljka Tasic, Kirsten Crichton, Josef Sulc, Boaz P. Levi, and Cindy T. J. van Velthoven
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Transcriptome ,Cell type ,Glutamatergic ,Neocortex ,medicine.anatomical_structure ,Taxonomy (general) ,medicine ,Hippocampus ,Hippocampal formation ,Biology ,Neuroscience ,Function (biology) - Abstract
The isocortex and hippocampal formation are two major structures in the mammalian brain that play critical roles in perception, cognition, emotion and learning. Using single-cell RNA-sequencing approaches, we profiled ~1.2 million cells covering all regions in the adult mouse isocortex and hippocampal formation. The cell types are organized hierarchically and exhibit varying degrees of discrete or continuous variations. Such molecular relationships correlate strongly with the spatial distribution patterns of the cell types, which can be region-specific, shared across multiple regions, or part of one or more gradients. Glutamatergic neuron types display much greater diversity than GABAergic neuron types, both molecularly and spatially, and define regional identities as well as inter-region relationships. Our study establishes a molecular architecture of the mammalian isocortex and hippocampal formation for the first time, and begins to shed light on its underlying relationship with the development, evolution, connectivity and function of these two brain structures.
- Published
- 2020
23. Enteroendocrine cells switch hormone expression along the crypt-to-villus BMP signalling gradient
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Es Johan H Van, Yorick Post, Thuc Nghi Nguyen, Benedetta Artegiani, Hans Clevers, Hongkui Zeng, Joep Beumer, den Born Maaike Van, Fiona M. Gribble, Frank Reimann, Hubrecht Institute for Developmental Biology and Stem Cell Research, Reimann, Frank [0000-0001-9399-6377], Gribble, Fiona [0000-0002-4232-2898], and Apollo - University of Cambridge Repository
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0301 basic medicine ,Enteroendocrine Cells ,Transgene ,Crypt ,Mice, Transgenic ,Enteroendocrine cell ,Bone Morphogenetic Protein 4 ,Biology ,digestive system ,Secretin ,Gastrointestinal Hormones ,Tissue Culture Techniques ,03 medical and health sciences ,Cell Movement ,Intestine, Small ,Animals ,Humans ,Cell Lineage ,LGR5 ,Cell Differentiation ,Cell Biology ,Phenotype ,Cell biology ,Mice, Inbred C57BL ,stomatognathic diseases ,Signalling ,030104 developmental biology ,MRNA Sequencing ,Signal Transduction ,Hormone - Abstract
Enteroendocrine cells (EECs) control a wide range of physiological processes linked to metabolism1. We show that EEC hormones are differentially expressed between crypts (for example, Glp1) and villi (for example, secretin). As demonstrated by single-cell mRNA sequencing using murine Lgr5+ cell-derived organoids, BMP4 signals alter the hormone expression profiles of individual EECs to resemble those found in the villus. Accordingly, BMP4 induces hormone switching of EECs migrating up the crypt-villus axis in vivo. Our findings imply that EEC lineages in the small intestine exhibit a more flexible hormone repertoire than previously proposed. We also describe a protocol to generate human EECs in organoids and demonstrate a similar regulation of hormone expression by BMP signalling. These findings establish alternative strategies to target EECs with therapeutically relevant hormone production through BMP modulation.
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- 2018
24. Cell segmentation-free inference of cell types from in situ transcriptomics data
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Oliver Stegle, Lars E. Borm, Sebastian Tiesmeyer, Wonyl Choi, Jeongbin Park, Emma Garren, Matthias Schlesner, Naveed Ishaque, Thuc Nghi Nguyen, Brian Long, Tobias Graf, Roland Eils, Simone Codeluppi, and Bosiljka Tasic
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In situ ,Cell type ,medicine.diagnostic_test ,Computer science ,Cell ,Cell segmentation ,Inference ,Computational biology ,Brain tissue ,Transcriptome ,medicine.anatomical_structure ,Visual cortex ,medicine ,Fluorescence in situ hybridization - Abstract
Summary Multiplexed fluorescence in situ hybridization techniques have enabled cell class or type identification by mRNA quantification in situ . However, inaccurate cell segmentation can result in incomplete cell-type and tissue characterization. Here, we present a robust segmentation-free computational framework, applicable to a variety of in situ transcriptomics platforms, called Spot-based Spatial cell-type Analysis by Multidimensional mRNA density estimation (SSAM). SSAM assumes that spatial distribution of mRNAs relates to organization of higher complexity structures (e.g. cells or tissue layers) and performs de novo cell-type and tissue domain identification. Optionally, SSAM can also integrate prior knowledge of cell types. We apply SSAM to three mouse brain tissue images: the somatosensory cortex imaged by osmFISH, the hypothalamic preoptic region by MERFISH, and the visual cortex by multiplexed smFISH. SSAM outperforms segmentation-based results, demonstrating that segmentation of cells is not required for inferring cell-type signatures, cell-type organization or tissue domains.
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- 2019
25. A taxonomy of transcriptomic cell types across the isocortex and hippocampal formation
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Zizhen Yao, Olivia Fong, Thanh Pham, Katelyn Ward, James Gray, Susan M. Sunkin, Stephanie Mok, Hongkui Zeng, Songlin Ding, Boaz P. Levi, Qingzhong Ren, Daniel Hirschstein, Emma Garren, Nick Dee, Megan Chiang, Fahimeh Baftizadeh, Christine Rimorin, Kanan Lathia, Herman Tung, Cindy T. J. van Velthoven, Darren Bertagnolli, Nadiya V. Shapovalova, Lucas T. Graybuck, Jeff Goldy, Michael Tieu, Delissa McMillen, Kimberly A. Smith, Michael Hawrylycz, Bosiljka Tasic, Amy Torkelson, Kirsten Crichton, Josef Sulc, Alexandra Glandon, Nathan W. Gouwens, Thuc Nghi Nguyen, Tamara Casper, Matthew Kroll, Adriana E. Sedeno-Cortes, and Changkyu Lee
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Cell type ,Interneuron ,Glutamic Acid ,Hippocampus ,Mice, Transgenic ,Neocortex ,Hippocampal formation ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Glutamatergic ,0302 clinical medicine ,Cortex (anatomy) ,medicine ,Animals ,GABAergic Neurons ,030304 developmental biology ,0303 health sciences ,Subiculum ,Mice, Inbred C57BL ,medicine.anatomical_structure ,GABAergic ,Transcriptome ,Neuroscience ,030217 neurology & neurosurgery - Abstract
The isocortex and hippocampal formation (HPF) in the mammalian brain play critical roles in perception, cognition, emotion, and learning. We profiled ∼1.3 million cells covering the entire adult mouse isocortex and HPF and derived a transcriptomic cell-type taxonomy revealing a comprehensive repertoire of glutamatergic and GABAergic neuron types. Contrary to the traditional view of HPF as having a simpler cellular organization, we discover a complete set of glutamatergic types in HPF homologous to all major subclasses found in the six-layered isocortex, suggesting that HPF and the isocortex share a common circuit organization. We also identify large-scale continuous and graded variations of cell types along isocortical depth, across the isocortical sheet, and in multiple dimensions in hippocampus and subiculum. Overall, our study establishes a molecular architecture of the mammalian isocortex and hippocampal formation and begins to shed light on its underlying relationship with the development, evolution, connectivity, and function of these two brain structures.
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- 2021
26. Conserved cell types with divergent features in human versus mouse cortex
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Elliot R. Thomsen, Ahmed Mahfouz, Saroja Somasundaram, Aaron Oldre, Bosiljka Tasic, Songlin Ding, Richard H. Scheuermann, Daniel Hirschstein, Thomas Höllt, Christine Rimorin, Thuc Nghi Nguyen, Jennie L. Close, John W. Phillips, Lydia Ng, Jeff Goldy, Darren Bertagnolli, Amy Bernard, Zizhen Yao, Boaz P. Levi, Trygve E. Bakken, Soraya I. Shehata, Susan M. Sunkin, Osnat Penn, Michael Tieu, Allison Beller, Boudewijn P. F. Lelieveldt, Jeffrey G. Ojemann, Shannon Reynolds, Michael Hawrylycz, Jeroen Eggermont, Medea McGraw, Ryder P. Gwinn, Sheana Parry, Kimberly A. Smith, Brian Long, Olivia Fong, Zoe Maltzer, Rafael Yuste, David Feng, Julie Nyhus, Rebecca D. Hodge, Ed Lein, Jeremy A. Miller, Brian D. Aevermann, Gerald Quon, Emma Garren, Christof Koch, Aaron Szafer, Nick Dee, Nadiya V. Shapovalova, Rachel A. Dalley, Tamara Casper, Mohamed Keshk, Nelson Johansen, Krissy Brouner, Andrew L. Ko, Allan R. Jones, Eliza Barkan, Hongkui Zeng, Richard G. Ellenbogen, C. Dirk Keene, Kanan Lathia, Lucas T. Graybuck, and Charles Cobbs
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0301 basic medicine ,Adult ,Male ,Cell type ,Adolescent ,General Science & Technology ,Middle temporal gyrus ,1.1 Normal biological development and functioning ,Biology ,03 medical and health sciences ,Mice ,Young Adult ,0302 clinical medicine ,Single-cell analysis ,Species Specificity ,Underpinning research ,Cortex (anatomy) ,medicine ,Genetics ,Animals ,Humans ,2.1 Biological and endogenous factors ,RNA-Seq ,Aetiology ,Aged ,Cerebral Cortex ,Neurons ,Principal Component Analysis ,Multidisciplinary ,Cellular architecture ,Neurosciences ,Neural Inhibition ,Human brain ,Middle Aged ,Biological Evolution ,030104 developmental biology ,medicine.anatomical_structure ,Cerebral cortex ,Astrocytes ,Neurological ,Excitatory postsynaptic potential ,Female ,Single-Cell Analysis ,Transcriptome ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Elucidating the cellular architecture of the human cerebral cortex is central to understanding our cognitive abilities and susceptibility to disease. Here we used single-nucleus RNA-sequencing analysis to perform a comprehensive study of cell types in the middle temporal gyrus of human cortex. We identified a highly diverse set of excitatory and inhibitory neuron types that are mostly sparse, with excitatory types being less layer-restricted than expected. Comparison to similar mouse cortex single-cell RNA-sequencing datasets revealed a surprisingly well-conserved cellular architecture that enables matching of homologous types and predictions of properties of human cell types. Despite this general conservation, we also found extensive differences between homologous human and mouse cell types, including marked alterations in proportions, laminar distributions, gene expression and morphology. These species-specific features emphasize the importance of directly studying human brain.
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- 2019
27. Multimodal cell type correspondence by intersectional mFISH in intact tissues
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Brian Long, Thuc Nghi Nguyen, Bosiljka Tasic, Boaz P. Levi, Ed S. Lein, Jeremy A. Miller, Jennie L. Close, Christopher A. Baker, Hongkui Zeng, Philip R. Nicovich, Alice Bosma-Moody, M. J. Taormina, Elliot R. Thomsen, Melissa Gorham, Emma Garren, Gabe J. Murphy, and Travis A. Hage
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Cell type ,medicine.anatomical_structure ,Mouse cortex ,Two-photon excitation microscopy ,medicine.diagnostic_test ,Cortex (anatomy) ,medicine ,Brain tissue ,Optogenetics ,Biology ,Correspondence problem ,Neuroscience ,Fluorescence in situ hybridization - Abstract
Defining a complete set of cell types within the cortex requires reconciling disparate results achieved through diverging methodologies. To address this correspondence problem, multiple methodologies must be applied to the same cells across multiple single-cell experiments. Here we present a new approach applying spatial transcriptomics using multiplexed fluorescencein situhybridization, (mFISH) to brain tissue previously interrogated through two photon optogenetic mapping of synaptic connectivity. This approach can resolve the anatomical, transcriptomic, connectomic, electrophysiological, and morphological characteristics of single cells within the mouse cortex.
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- 2019
28. Complete Whole-Brain Single Neuron Reconstruction Reveals Morphological Diversity in Molecularly Defined Claustral and Cortical Neuron Types
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Z. Josh Huang, Matthew B. Veldman, Yang Yu, Sara Kebede, Philip Lesnar, Thuc Nghi Nguyen, Anan Li, Zhi Zhou, Chris Hill, Susan M. Sunkin, Hanchuan Peng, Shaoqun Zeng, Yaoyao Li, Qingming Luo, Li Xiangning, Lei Qu, Karla E. Hirokawa, Elise Shen, Lijuan Liu, Yimin Wang, X. William Yang, Michael Hawrylycz, Lydia Ng, Hui Gong, Peng Xie, Sujun Zhao, Xiuli Kuang, Tanya L. Daigle, Aaron Feiner, Zizhen Yao, Christof Koch, Shengdian Jiang, Stephanie Mok, Jing Yuan, Hongkui Zeng, Lulu Ying, Rachael Larsen, Staci A. Sorensen, Julie A. Harris, Luke Esposito, Yun Wang, Bosiljka Tasic, Yuanyuan Song, Quanxin Wang, and Wayne Wakeman
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Fluorescent labelling ,medicine.anatomical_structure ,Feature (computer vision) ,Cortical neuron ,Cortex (anatomy) ,medicine ,Neuron ,Biology ,Fluorescent imaging ,Neuroscience ,Claustrum - Abstract
Dendritic and axonal morphology is a defining feature of neuronal types and their connectivity. Yet our knowledge concerning the diversity of neuronal morphology is extremely limited. To systematically obtain single neuron full morphology on a brain-wide scale in mice, we established a pipeline that encompasses five major components: sparse labeling, whole-brain imaging, reconstruction, registration, and classification. We achieved sparse, robust and consistent fluorescent labeling by combining transgenic or viral Cre delivery with novel transgenic reporter lines, and generated whole-brain fluorescent imaging datasets containing tens of thousands of reconstructable neurons. We developed software tools for large-volume image data processing and computer-assisted morphological reconstruction. For a proof-of-principle, we reconstructed the full morphologies of 96 neurons from the claustrum and cortex that belong to a single transcriptomically-defined subclass, and classified them into multiple morphological types, suggesting that they work in a targeted and coordinated manner to process cortical information over a large region.
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- 2019
29. Single-nucleus and single-cell transcriptomes compared in matched cortical cell types
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Jeremy A. Miller, Soraya I. Shehata, Thuc Nghi Nguyen, Matthew Kroll, Nick Dee, Sheana Parry, Rebecca D. Hodge, Kimberly A. Smith, Brian D. Aevermann, Ed Lein, Christine Rimorin, Darren Bertagnolli, Amy Bernard, Michael Tieu, Bosiljka Tasic, Eliza Barkan, Jeff Goldy, Emma Garren, Tamara Casper, Richard H. Scheuermann, Hongkui Zeng, Nicholas J. Schork, Trygve E. Bakken, Roger S. Lasken, Kanan Lathia, John W. Phillips, Lucas T. Graybuck, and Zizhen Yao
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0301 basic medicine ,Cell ,genetic processes ,Gene Expression ,Biochemistry ,Transcriptome ,Database and Informatics Methods ,Mice ,Single-cell analysis ,Animal Cells ,Gene expression ,Visual Cortex ,Neurons ,Multidisciplinary ,Mammalian Genomics ,Messenger RNA ,Genomics ,Cell biology ,Nucleic acids ,medicine.anatomical_structure ,Medicine ,Cellular Types ,Single-Cell Analysis ,Transcriptome Analysis ,Sequence Analysis ,Research Article ,Cell type ,Sequence analysis ,Bioinformatics ,Science ,Biology ,Research and Analysis Methods ,Genome Complexity ,03 medical and health sciences ,medicine ,Genetics ,Animals ,natural sciences ,Cell Lineage ,Molecular Biology Techniques ,Molecular Biology ,Cell Nucleus ,Sequence Analysis, RNA ,Gene Expression Profiling ,Intron ,Biology and Life Sciences ,Computational Biology ,Marker Genes ,Cell Biology ,Genome Analysis ,Introns ,030104 developmental biology ,Animal Genomics ,Cellular Neuroscience ,RNA ,Nucleus ,Sequence Alignment ,Neuroscience - Abstract
Transcriptomic profiling of complex tissues by single-nucleus RNA-sequencing (snRNA-seq) affords some advantages over single-cell RNA-sequencing (scRNA-seq). snRNA-seq provides less biased cellular coverage, does not appear to suffer cell isolation-based transcriptional artifacts, and can be applied to archived frozen specimens. We used well-matched snRNA-seq and scRNA-seq datasets from mouse visual cortex to compare cell type detection. Although more transcripts are detected in individual whole cells (~11,000 genes) than nuclei (~7,000 genes), we demonstrate that closely related neuronal cell types can be similarly discriminated with both methods if intronic sequences are included in snRNA-seq analysis. We estimate that the nuclear proportion of total cellular mRNA varies from 20% to over 50% for large and small pyramidal neurons, respectively. Together, these results illustrate the high information content of nuclear RNA for characterization of cellular diversity in brain tissues.
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- 2018
30. Single-cell and single-nucleus RNA-seq uncovers shared and distinct axes of variation in dorsal LGN neurons in mice, non-human primates, and humans.
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Bakken, Trygve E., van Velthoven, Cindy T. J., Meno, Vilas, Hodge, Rebecca D., Zizhen Yao, Thuc Nghi Nguyen, Graybuck, Lucas T., Horwitz, Gregory D., Bertagnolli, Darren, Goldy, Jeff, Yanny, Anna Marie, Garren, Emma, Parry, Sheana, Casper, Tamara, Shehata, Soraya I., Barkan, Eliza R., Szafer, Aaron, Levi, Boaz P., Dee, Nick, and Smith, Kimberly A.
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- 2021
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31. Adult Mouse Cortical Cell Taxonomy by Single Cell Transcriptomics
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Linda Madisen, Kimberly A. Smith, Darren Bertagnolli, Amy Bernard, Tim Jarsky, Jeff Goldy, Tim A. Dolbeare, Hongkui Zeng, Lucas T. Gray, Christof Koch, Tae Kyung Kim, Susan M. Sunkin, Boaz P. Levi, Changkyu Lee, Bosiljka Tasic, Nadiya V. Shapovalova, Zizhen Yao, Michael Hawrylycz, Staci A. Sorensen, Thuc Nghi Nguyen, Sheana Parry, and Vilas Menon
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0301 basic medicine ,Genetic Markers ,Male ,Cell type ,Transgene ,Cell ,Glutamic Acid ,Mice, Transgenic ,Biology ,Article ,Cell Line ,Transcriptome ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Interneurons ,medicine ,Animals ,Axon ,gamma-Aminobutyric Acid ,Gene Library ,Visual Cortex ,Neurons ,Cerebral Cortex ,Sequence Analysis, RNA ,General Neuroscience ,Classification ,Molecular biology ,Cell biology ,Gene expression profiling ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Cerebral cortex ,Cell culture ,RNA ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Nervous systems are composed of various cell types, but the extent of cell type diversity is poorly understood. We constructed a cellular taxonomy of one cortical region, primary visual cortex, in adult mice on the basis of single-cell RNA sequencing. We identified 49 transcriptomic cell types, including 23 GABAergic, 19 glutamatergic and 7 non-neuronal types. We also analyzed cell type-specific mRNA processing and characterized genetic access to these transcriptomic types by many transgenic Cre lines. Finally, we found that some of our transcriptomic cell types displayed specific and differential electrophysiological and axon projection properties, thereby confirming that the single-cell transcriptomic signatures can be associated with specific cellular properties.
- Published
- 2016
32. Distinct Transcriptomic Cell Types and Neural Circuits of the Subiculum and Prosubiculum along the Dorsal-Ventral Axis
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Kimberly A. Smith, Julie A. Harris, Thuc Nghi Nguyen, Karla E. Hirokawa, Phillip Bohn, Kiet Ngo, Lucas T. Graybuck, Ed Lein, John W. Phillips, Bosiljka Tasic, Hongkui Zeng, Christof Koch, Olivia Fong, Zizhen Yao, and Songlin Ding
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0301 basic medicine ,Cell type ,Future studies ,Biology ,Hippocampus ,Article ,General Biochemistry, Genetics and Molecular Biology ,Transcriptome ,Dorsal ventral ,03 medical and health sciences ,0302 clinical medicine ,Neural Pathways ,scRNA-seq ,Biological neural network ,Animals ,subicular complex ,Functional studies ,lcsh:QH301-705.5 ,prosubiculum ,Functional correlation ,Subiculum ,Entorhinal cortex ,030104 developmental biology ,lcsh:Biology (General) ,Prosubiculum ,single-cell transcriptomics ,cell types ,ventral hippocampus ,Neuroscience ,030217 neurology & neurosurgery - Abstract
SummarySubicular region plays important roles in spatial processing and many cognitive functions and these were mainly attributed to subiculum (Sub) rather than prosubiculum (PS). Using single-cell RNA-sequencing (scRNA-seq) technique we have identified up to 27 distinct transcriptomic clusters/cell types, which were registered to anatomical sub-domains in Sub and PS. Based on reliable molecular markers derived from transcriptomic clustering and in situ hybridization data, the precise boundaries of Sub and PS have been consistently defined along the dorsoventral (DV) axis. Using these borders to evaluate Cre-line specificity and tracer injections, we have found bona fide Sub projections topographically to structures important for spatial processing and navigation. In contrast, PS along DV axis sends its outputs to widespread brain regions crucial for motivation, emotion, reward, stress, anxiety and fear. Brain-wide cell-type specific projections of Sub and PS have also been revealed using specific Cre-lines. These results reveal two molecularly and anatomically distinct circuits centered in Sub and PS, respectively, providing a consistent explanation to historical data and a clearer foundation for future functional studies.Highlights27 transcriptomic cell types identified in and spatially registered to “subicular” regions.Anatomic borders of “subicular” regions reliably determined along dorsal-ventral axis.Distinct cell types and circuits of full-length subiculum (Sub) and prosubiculum (PS).Brain-wide cell-type specific projections of Sub and PS revealed with specific Cre-lines.In BriefDing et al. show that mouse subiculum and prosubiculum are two distinct regions with differential transcriptomic cell types, subtypes, neural circuits and functional correlation. The former has obvious topographic projections to its main targets while the latter exhibits widespread projections to many subcortical regions associated with reward, emotion, stress and motivation.
- Published
- 2020
33. Conserved cell types with divergent features between human and mouse cortex
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Jeff Goldy, Sheana Parry, Jeremy A. Miller, Brian Long, Susan M. Sunkin, Saroja Somasundaram, Rebecca D. Hodge, Hongkui Zeng, Aaron Oldre, Kimberly A. Smith, Zoe Maltzer, Brian D. Aevermann, Mohamed Keshk, Jeroen Eggermont, Ed Lein, Daniel Hirschstein, Darren Bertagnolli, Jennie L. Close, Osnat Penn, John W. Phillips, Rachel A. Dalley, Allan R. Jones, Ahmed Mahfouz, Olivia Fong, Allison Beller, Soraya I. Shehata, Thuc Nghi Nguyen, Jeffrey G. Ojemann, Shannon Reynolds, Eliza Barkan, Michael Tieu, Christof Koch, Michael Hawrylycz, Songlin Ding, Richard H. Scheuermann, Ryder P. Gwinn, Elliot R. Thomsen, Medea McGraw, Emma Garren, Christine Rimorin, Lydia Ng, Boudewijn P. F. Lelieveldt, C. Dirk Keene, Amy Bernard, Richard G. Ellenbogen, Rafael Yuste, David Feng, Boaz P. Levi, Trygve E. Bakken, Tamara Casper, Bosiljka Tasic, Aaron Szafer, Nick Dee, Nadiya V. Shapovalova, Kanan Lathia, Lucas T. Graybuck, Charles Cobbs, Julie Nyhus, Thomas Höllt, Zizhen Yao, Krissy Brouner, and Andrew L. Ko
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0303 health sciences ,Cell type ,Neocortex ,Cellular architecture ,Middle temporal gyrus ,Cell ,Human brain ,Biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Cerebral cortex ,medicine ,Neuroscience ,Nucleus ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
Elucidating the cellular architecture of the human neocortex is central to understanding our cognitive abilities and susceptibility to disease. Here we applied single nucleus RNA-sequencing to perform a comprehensive analysis of cell types in the middle temporal gyrus of human cerebral cortex. We identify a highly diverse set of excitatory and inhibitory neuronal types that are mostly sparse, with excitatory types being less layer-restricted than expected. Comparison to a similar mouse cortex single cell RNA-sequencing dataset revealed a surprisingly well-conserved cellular architecture that enables matching of homologous types and predictions of human cell type properties. Despite this general conservation, we also find extensive differences between homologous human and mouse cell types, including dramatic alterations in proportions, laminar distributions, gene expression, and morphology. These species-specific features emphasize the importance of directly studying human brain.
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- 2018
34. Classification of electrophysiological and morphological types in mouse visual cortex
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Tom Egdorf, Rebecca de Frates, Emma Garren, Sara Kebede, Peter Chong, John W. Phillips, Nivretta Thatra, Samuel R Josephsen, Philip R. Nicovich, Tim Jarsky, Xiaoxiao Liu, Susan M. Sunkin, Brian Lee, Keith B. Godfrey, Matthew Kroll, Nicole Blesie, Bosiljka Tasic, Amy Bernard, Lisa Kim, Costas A. Anastassiou, Kristen Hadley, Staci A. Sorensen, Thuc Nghi Nguyen, Martin Schroedter, Corinne Teeter, Kirsten Crichton, Josef Sulc, Rachel A. Dalley, David Feng, Tracy Lemon, Michael Hawrylycz, Miranda Robertson, Christine Cuhaciyan, Eliza Barkan, Shiella Caldejon, Tsega Desta, Kris Bickley, Dan Castelli, Wayne Wakeman, Herman Tung, Hongkui Zeng, Grace Williams, Nadezhda Dotson, Rusty Mann, Tamara Casper, Anton Arkhipov, Daniel Park, Sheana Parry, Jed Perkins, Alyse Doperalski, Brian Long, Thomas Braun, Christof Koch, Gabe J. Murphy, Aaron Oldre, Changkyu Lee, Colin Farrell, Medea McGraw, Amanda Gary, Kiet Ngo, Melissa Gorham, Naz Taskin, Jim Berg, Samuel Dingman, Tanya L. Daigle, Agata Budzillo, Marissa Garwood, Gilberto J. Soler-Llavina, Aaron Szafer, Nick Dee, Jonathan T. Ting, Lydia Ng, Alex M. Henry, James Harrington, Julie A. Harris, Michael S. Fisher, Lindsay Ng, Caroline Habel, Nathalie Gaudreault, Krissy Brouner, David Reid, Lydia Potekhina, Rob Young, DiJon Hill, Cliff Slaughterbeck, Ed Lein, Alice Mukora, David Sandman, Stefan Mihalas, Nathan W. Gouwens, Zhi Zhou, Hanchuan Peng, and Hong Gu
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Genetically modified mouse ,Cell type ,Cell ,Laboratory mouse ,Biology ,Electrophysiology ,chemistry.chemical_compound ,Visual cortex ,medicine.anatomical_structure ,chemistry ,Biocytin ,medicine ,Patch clamp ,Neuroscience - Abstract
Understanding the diversity of cell types in the brain has been an enduring challenge and requires detailed characterization of individual neurons in multiple dimensions. To profile morpho-electric properties of mammalian neurons systematically, we established a single cell characterization pipeline using standardized patch clamp recordings in brain slices and biocytin-based neuronal reconstructions. We built a publicly-accessible online database, the Allen Cell Types Database, to display these data sets. Intrinsic physiological and morphological properties were measured from over 1,800 neurons from the adult laboratory mouse visual cortex. Quantitative features were used to classify neurons into distinct types using unsupervised methods. We establish a taxonomy of morphologically- and electrophysiologically-defined cell types for this region of cortex with 17 e-types and 35 m-types, as well as an initial correspondence with previously-defined transcriptomic cell types using the same transgenic mouse lines.
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- 2018
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35. Multimodal Analysis of Cell Types in a Hypothalamic Node Controlling Social Behavior
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Tae Kyung Kim, Dong-Wook Kim, David J. Anderson, Lucas T. Graybuck, Lior Pachter, Thuc Nghi Nguyen, Lynn Yi, Sheel Shah, Yuki Oka, Long Cai, Liching Lo, Zizhen Yao, Kimberly A. Smith, Bosiljka Tasic, Nico Pierson, Olivia Fong, Noushin Koulena, Allan-Hermann Pool, and Hongkui Zeng
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Male ,Cell type ,Population ,Hypothalamus ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,Sexual Behavior, Animal ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Social Behavior ,education ,030304 developmental biology ,Neurons ,Mice, Inbred BALB C ,0303 health sciences ,education.field_of_study ,Aggression ,Estrogen Receptor alpha ,Behavioral activation ,Mice, Inbred C57BL ,Sexual dimorphism ,Female ,Single-Cell Analysis ,medicine.symptom ,Transcriptome ,Neuroscience ,Immediate early gene ,030217 neurology & neurosurgery ,Social behavior - Abstract
The ventrolateral subdivision of the ventromedial hypothalamus (VMHvl) contains ∼4,000 neurons that project to multiple targets and control innate social behaviors including aggression and mounting. However, the number of cell types in VMHvl and their relationship to connectivity and behavioral function are unknown. We performed single-cell RNA sequencing using two independent platforms-SMART-seq (∼4,500 neurons) and 10x (∼78,000 neurons)-and investigated correspondence between transcriptomic identity and axonal projections or behavioral activation, respectively. Canonical correlation analysis (CCA) identified 17 transcriptomic types (T-types), including several sexually dimorphic clusters, the majority of which were validated by seqFISH. Immediate early gene analysis identified T-types exhibiting preferential responses to intruder males versus females but only rare examples of behavior-specific activation. Unexpectedly, many VMHvl T-types comprise a mixed population of neurons with different projection target preferences. Overall our analysis revealed that, surprisingly, few VMHvl T-types exhibit a clear correspondence with behavior-specific activation and connectivity.
- Published
- 2019
36. Shared and distinct transcriptomic cell types across neocortical areas
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Daniel Hirschstein, Michael N. Economo, Allan R. Jones, Christine Rimorin, Eliza Barkan, Linda Madisen, Seana Parry, Susan M. Sunkin, Rachael Larsen, Hongkui Zeng, Tae Kyung Kim, Emma Garren, Kimberly A. Smith, Jeremy A. Miller, Osnat Penn, Olivia Fong, Sarada Viswanathan, Julie A. Harris, Bosiljka Tasic, Thuc Nghi Nguyen, Vilas Menon, Karel Svoboda, Matthew Kroll, Ed S. Lein, Peter A. Groblewski, Karla E. Hirokawa, Ali Cetin, Julie Pendergraft, Ian R. Wickersham, Tanya L. Daigle, Darren Bertagnolli, Jeff Goldy, Zizhen Yao, John W. Phillips, Michael Tieu, Loren L. Looger, Michael Hawrylycz, Aaron Szafer, Boaz P. Levi, Trygve E. Bakken, Nick Dee, Nadiya V. Shapovalova, Amy Bernard, Tamara Casper, Christof Koch, Kanan Lathia, and Lucas T. Graybuck
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Transcriptome ,Cell type ,medicine.anatomical_structure ,Neocortex ,Visual cortex ,Cell ,medicine ,Excitatory postsynaptic potential ,Biology ,Inhibitory postsynaptic potential ,Neuroscience ,Motor cortex - Abstract
Neocortex contains a multitude of cell types segregated into layers and functionally distinct regions. To investigate the diversity of cell types across the mouse neocortex, we analyzed 12,714 cells from the primary visual cortex (VISp), and 9,035 cells from the anterior lateral motor cortex (ALM) by deep single-cell RNA-sequencing (scRNA-seq), identifying 116 transcriptomic cell types. These two regions represent distant poles of the neocortex and perform distinct functions. We define 50 inhibitory transcriptomic cell types, all of which are shared across both cortical regions. In contrast, 49 of 52 excitatory transcriptomic types were found in either VISp or ALM, with only three present in both. By combining single cell RNA-seq and retrograde labeling, we demonstrate correspondence between excitatory transcriptomic types and their region-specific long-range target specificity. This study establishes a combined transcriptomic and projectional taxonomy of cortical cell types from functionally distinct regions of the mouse cortex.
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- 2017
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37. Shared and distinct transcriptomic cell types across neocortical areas
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Zizhen Yao, Christof Koch, Daniel Hirschstein, Aaron Szafer, Nick Dee, Sheana Parry, Michael Tieu, Michael Hawrylycz, Jeff Goldy, Susan M. Sunkin, Nadiya V. Shapovalova, Amy Bernard, Kanan Lathia, Lucas T. Graybuck, Boaz P. Levi, Trygve E. Bakken, Matthew Kroll, Sarada Viswanathan, Kimberly A. Smith, Thuc Nghi Nguyen, Olivia Fong, Tae Kyung Kim, Tanya L. Daigle, Jeremy A. Miller, Christine Rimorin, Linda Madisen, Karla E. Hirokawa, Tamara Casper, Julie A. Harris, Ali Cetin, Heather A. Sullivan, Bosiljka Tasic, Karel Svoboda, Julie Pendergraft, Osnat Penn, John W. Phillips, Ian R. Wickersham, Ed S. Lein, Loren L. Looger, Peter A. Groblewski, Hongkui Zeng, Allan R. Jones, Rachael Larsen, Emma Garren, Darren Bertagnolli, Michael N. Economo, Eliza Barkan, and Vilas Menon
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0301 basic medicine ,Male ,Cell type ,Glutamic Acid ,Neocortex ,Biology ,Article ,Transcriptome ,03 medical and health sciences ,Glutamatergic ,Mice ,Single-cell analysis ,medicine ,Animals ,GABAergic Neurons ,Visual Cortex ,Multidisciplinary ,Sequence Analysis, RNA ,Gene Expression Profiling ,Motor Cortex ,Gene expression profiling ,030104 developmental biology ,medicine.anatomical_structure ,Visual cortex ,nervous system ,Organ Specificity ,Female ,Single-Cell Analysis ,Neuroscience ,Biomarkers ,Motor cortex - Abstract
The neocortex contains a multitude of cell types that are segregated into layers and functionally distinct areas. To investigate the diversity of cell types across the mouse neocortex, here we analysed 23,822 cells from two areas at distant poles of the mouse neocortex: the primary visual cortex and the anterior lateral motor cortex. We define 133 transcriptomic cell types by deep, single-cell RNA sequencing. Nearly all types of GABA (γ-aminobutyric acid)-containing neurons are shared across both areas, whereas most types of glutamatergic neurons were found in one of the two areas. By combining single-cell RNA sequencing and retrograde labelling, we match transcriptomic types of glutamatergic neurons to their long-range projection specificity. Our study establishes a combined transcriptomic and projectional taxonomy of cortical cell types from functionally distinct areas of the adult mouse cortex.
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- 2017
38. Distinct descending motor cortex pathways and their roles in movement
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Erhan Bas, Lihua Wang, Thuc Nghi Nguyen, Johan Winnubst, Hongkui Zeng, Vilas Menon, Loren L. Looger, Charles R. Gerfen, Bosiljka Tasic, Karel Svoboda, Lucas T. Graybuck, Jayaram Chandrashekar, Sarada Viswanathan, and Michael N. Economo
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Pyramidal tracts ,medicine.anatomical_structure ,Basal ganglia ,Motor commands ,medicine ,Motor control ,Brainstem ,Neuron ,Biology ,Neuroscience ,Medulla ,Motor cortex - Abstract
Activity in motor cortex predicts specific movements, seconds before they are initiated. This preparatory activity has been observed in L5 descending ‘pyramidal tract’ (PT) neurons. A key question is how preparatory activity can be maintained without causing movement, and how preparatory activity is eventually converted to a motor command to trigger appropriate movements. We used single cell transcriptional profiling and axonal reconstructions to identify two types of PT neuron. Both types share projections to multiple targets in the basal ganglia and brainstem. One type projects to thalamic regions that connect back to motor cortex. In a delayed-response task, these neurons produced early preparatory activity that persisted until the movement. The second type projects to motor centers in the medulla and produced late preparatory activity and motor commands. These results indicate that two motor cortex output neurons are specialized for distinct roles in motor control.
- Published
- 2017
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39. Author response: Layer-specific chromatin accessibility landscapes reveal regulatory networks in adult mouse visual cortex
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Zizhen Yao, Lucas T. Gray, Thuc Nghi Nguyen, Hongkui Zeng, Tae Kyung Kim, and Bosiljka Tasic
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Visual cortex ,medicine.anatomical_structure ,medicine ,Layer (object-oriented design) ,Biology ,Neuroscience ,Chromatin - Published
- 2017
40. Layer-specific chromatin accessibility landscapes reveal regulatory networks in adult mouse visual cortex
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Tae Kyung Kim, Zizhen Yao, Lucas T. Gray, Thuc Nghi Nguyen, Hongkui Zeng, and Bosiljka Tasic
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0301 basic medicine ,Cell type ,Mouse ,QH301-705.5 ,Science ,Transposases ,Computational biology ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Gene Regulatory Networks ,visual cortex ,Biology (General) ,Gene ,Transcription factor ,ChIA-PET ,Neurons ,Genetics ,General Immunology and Microbiology ,Gene Expression Profiling ,General Neuroscience ,High-Throughput Nucleotide Sequencing ,FOXP2 ,General Medicine ,ATAC-seq ,Chromatin ,Tools and Resources ,030104 developmental biology ,Visual cortex ,medicine.anatomical_structure ,Genomics and Evolutionary Biology ,NFIA ,chromatin accessibility ,Medicine ,transcription ,030217 neurology & neurosurgery ,Protein Binding ,Neuroscience - Abstract
Mammalian cortex is a laminar structure, with each layer composed of a characteristic set of cell types with different morphological, electrophysiological, and connectional properties. Here, we define chromatin accessibility landscapes of major, layer-specific excitatory classes of neurons, and compare them to each other and to inhibitory cortical neurons using the Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq). We identify a large number of layer-specific accessible sites, and significant association with genes that are expressed in specific cortical layers. Integration of these data with layer-specific transcriptomic profiles and transcription factor binding motifs enabled us to construct a regulatory network revealing potential key layer-specific regulators, including Cux1/2, Foxp2, Nfia, Pou3f2, and Rorb. This dataset is a valuable resource for identifying candidate layer-specific cis-regulatory elements in adult mouse cortex. DOI: http://dx.doi.org/10.7554/eLife.21883.001
- Published
- 2017
41. Identification of preoptic sleep neurons using retrograde labelling and gene profiling
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Franz Weber, Wei-Cheng Chang, Johnny Phong Hoang Do, Hongkui Zeng, Shinjae Chung, Thuc Nghi Nguyen, Nikolai Hörmann, Chan Lek Tan, Kevin T. Beier, Liqun Luo, Peng Zhong, Michael J. Krashes, Shenqin Yao, Bosiljka Tasic, Yang Dan, Zhe Zhang, Ali Cetin, and Zachary A. Knight
- Subjects
0301 basic medicine ,Male ,medicine.medical_specialty ,endocrine system ,Corticotropin-Releasing Hormone ,Population ,Channelrhodopsin ,Optogenetics ,Biology ,Article ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Channelrhodopsins ,Chloride Channels ,Internal medicine ,Tachykinins ,medicine ,Animals ,GABAergic Neurons ,Wakefulness ,education ,Neurons ,education.field_of_study ,Multidisciplinary ,Sequence Analysis, RNA ,Preoptic Area ,Preoptic area ,Neuroanatomical Tract-Tracing Techniques ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,nervous system ,Hypothalamus ,Hypothalamic Area, Lateral ,GABAergic ,Female ,Sleep onset ,Single-Cell Analysis ,Tuberomammillary nucleus ,Cholecystokinin ,Sleep ,Transcriptome ,Neuroscience ,Proto-Oncogene Proteins c-fos ,Ribosomes ,030217 neurology & neurosurgery ,Biomarkers - Abstract
Identification of sleep-active and sleep-promoting neurons in the preoptic area of the hypothalamus using neural projection tracing tools to target this population among a group of intermingled neurons, all with various functions. The preoptic area (POA) in the hypothalamus is an essential contributor to typical sleep regulation, but how this brain area is involved in this process has not been well-understood. Now, Yang Dan and colleagues dissect the role of sleep-active neurons in the POA using neural-projection-tracing tools to specifically target this population of neurons amongst a group of intermingled neurons with various functions. The POA sleep neurons were GABAergic and projected to the tuberomammillary nucleus and were not only active during sleep but could promote sleep when activated. Further, single-cell molecular analysis provided candidate genetic markers with which to target these neurons for future studies aiming to further dissect this sleep control circuit. In humans and other mammalian species, lesions in the preoptic area of the hypothalamus cause profound sleep impairment1,2,3,4,5, indicating a crucial role of the preoptic area in sleep generation. However, the underlying circuit mechanism remains poorly understood. Electrophysiological recordings and c-Fos immunohistochemistry have shown the existence of sleep-active neurons in the preoptic area, especially in the ventrolateral preoptic area and median preoptic nucleus6,7,8,9. Pharmacogenetic activation of c-Fos-labelled sleep-active neurons has been shown to induce sleep10. However, the sleep-active neurons are spatially intermingled with wake-active neurons6,7, making it difficult to target the sleep neurons specifically for circuit analysis. Here we identify a population of preoptic area sleep neurons on the basis of their projection target and discover their molecular markers. Using a lentivirus expressing channelrhodopsin-2 or a light-activated chloride channel for retrograde labelling, bidirectional optogenetic manipulation, and optrode recording, we show that the preoptic area GABAergic neurons projecting to the tuberomammillary nucleus are both sleep active and sleep promoting. Furthermore, translating ribosome affinity purification and single-cell RNA sequencing identify candidate markers for these neurons, and optogenetic and pharmacogenetic manipulations demonstrate that several peptide markers (cholecystokinin, corticotropin-releasing hormone, and tachykinin 1) label sleep-promoting neurons. Together, these findings provide easy genetic access to sleep-promoting preoptic area neurons and a valuable entry point for dissecting the sleep control circuit.
- Published
- 2016
42. A Suite of Transgenic Driver and Reporter Mouse Lines with Enhanced Brain-Cell-Type Targeting and Functionality
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Travis A. Hage, Christopher A. Baker, Linda Madisen, Alice Bosma-Moody, Rylan S. Larsen, Matthew T. Valley, Jonathan T. Ting, Karla E. Hirokawa, Kimberly A. Smith, Olivia Fong, Jérôme Lecoq, Garreck H. Lenz, Julie Pendergraft, Susan M. Sunkin, Julie A. Harris, La'Akea Siverts, Maya Mills, Zizhen Yao, Michael Z. Lin, Thuc Nghi Nguyen, Mariya Chavarha, Philip R. Nicovich, Nuno Maçarico da Costa, Lawrence Huang, Lu Li, Miranda Walker, Marc Takeno, Gabe J. Murphy, Lucas T. Graybuck, Jack Waters, Emma Garren, Edward S. Boyden, Medea McGraw, Rachael Larsen, James Harrington, Douglas R. Ollerenshaw, Ulf Knoblich, Tanya L. Daigle, Hongkui Zeng, Bosiljka Tasic, and Hong Gu
- Subjects
0301 basic medicine ,Genetically modified mouse ,Cell type ,RNA, Untranslated ,Light ,Transgene ,Cell ,Mice, Transgenic ,Computational biology ,Optogenetics ,Biology ,Brain Cell ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Gene Knockout Techniques ,Mice ,03 medical and health sciences ,Genes, Reporter ,health services administration ,medicine ,Animals ,natural sciences ,Transgenes ,In Situ Hybridization, Fluorescence ,Neurons ,Brain ,Transgenesis ,030104 developmental biology ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Calcium ,human activities ,Function (biology) - Abstract
Modern genetic approaches are powerful in providing access to diverse cell types in the brain and facilitating the study of their function. Here, we report a large set of driver and reporter transgenic mouse lines, including 23 new driver lines targeting a variety of cortical and subcortical cell populations and 26 new reporter lines expressing an array of molecular tools. In particular, we describe the TIGRE2.0 transgenic platform and introduce Cre-dependent reporter lines that enable optical physiology, optogenetics, and sparse labeling of genetically defined cell populations. TIGRE2.0 reporters broke the barrier in transgene expression level of single-copy targeted-insertion transgenesis in a wide range of neuronal types, along with additional advantage of a simplified breeding strategy compared to our first-generation TIGRE lines. These novel transgenic lines greatly expand the repertoire of high-precision genetic tools available to effectively identify, monitor, and manipulate distinct cell types in the mouse brain.
- Published
- 2018
43. Monte Carlo Simulation of Cell Death Signaling Predicts Large Cell-to-Cell Stochastic Fluctuations through the Type 2 Pathway of Apoptosis
- Author
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Eric Willgohs, Subhadip Raychaudhuri, Elaine M. Khan, Tzipora Goldkorn, and Thuc Nghi Nguyen
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Programmed cell death ,Cell signaling ,Time Factors ,Population ,Cell ,Biophysics ,Caspase 3 ,Apoptosis ,Biophysical Theory and Modeling ,Biology ,Bioinformatics ,Models, Biological ,Enzyme activator ,medicine ,Humans ,Computer Simulation ,education ,education.field_of_study ,Stochastic Processes ,Cell biology ,Enzyme Activation ,medicine.anatomical_structure ,Signal transduction ,Monte Carlo Method ,Signal Transduction - Abstract
Apoptosis, or genetically programmed cell death, is a crucial cellular process that maintains the balance between life and death in cells. The precise molecular mechanism of apoptosis signaling and the manner in which type 1 and type 2 pathways of the apoptosis signaling network are differentially activated under distinct apoptotic stimuli is poorly understood. Based on Monte Carlo stochastic simulations, we show that the type 1 pathway becomes activated under strong apoptotic stimuli, whereas the type 2 mitochondrial pathway dominates apoptotic signaling in response to a weak death signal. Our results also show signaling in the type 2 pathway is stochastic; the population average over many cells does not capture the cell-to-cell fluctuations in the time course (approximately 1-10 h) of downstream caspase-3 activation. On the contrary, the probability distribution of caspase-3 activation for the mitochondrial pathway shows a distinct bimodal behavior that can be used to characterize the stochastic signaling in type 2 apoptosis and other similar complex signaling processes. Interestingly, such stochastic fluctuations in apoptosis signaling occur even in the presence of large numbers of signaling molecules.
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- 2008
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44. Do Initial Radiographs Agree With Crash Site Mechanism of Injury in Pelvic Ring Disruptions? A Pilot Study
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Milton L Routt, C. Craig Blackmore, Gregory J. Jurkovich, Charles Mock, Robert Kaufman, Thuc Nghi Nguyen, Lloyd E. Stambaugh, and Ken F. Linnau
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Adult ,Male ,medicine.medical_specialty ,Databases, Factual ,Radiography ,Poison control ,Pilot Projects ,Crash ,medicine.disease_cause ,Weight-bearing ,Weight-Bearing ,Fractures, Bone ,Injury Severity Score ,Cohen's kappa ,Trauma Centers ,medicine ,Humans ,Orthopedics and Sports Medicine ,Pelvic Bones ,Observer Variation ,Orthodontics ,Pelvic girdle ,business.industry ,Accidents, Traffic ,Reproducibility of Results ,General Medicine ,medicine.disease ,Surgery ,Pelvic fracture ,Female ,Stress, Mechanical ,business - Abstract
Objective: Direction of injury force inferred from pelvic radiographs may be used in trauma care to predict associated injuries and guide intervention. Our objective was to compare injury direction determined from anteroposterior (AP) pelvic radiographs with injury forces determined from crash site investigation. Materials and Methods: We studied all 28 subjects from the Crash Injury Research Engineering Network (CIREN) database who met inclusion criteria of pelvic ring disruption, single-event crash, restrained front-seat occupant, diagnostic-quality pelvic radiography, and complete crash investigation data. Assessment of diagnostic quality of pelvic radiography was made by 2 radiologists who were blinded to all other subject information. Crash site investigation data included principal direction of force (PDOF), crash magnitude, and passenger compartment intrusion. An orthopedic trauma surgeon and a fellowship-trained emergency radiologist independently assessed the pelvic radiographs to determine the injury PDOF and the YoungBurgess and Tile fracture classifications, with disputes resolved by an additional emergency radiologist. Agreement between injury forces and pelvic radiographs was assessed using the kappa statistic. Results: The PDOF was anterior in 9 (32%) and lateral in 19 (68%) subjects. The readers agreed with the crash primary direction of force in 21 (75%) subjects (k = 0.42). In subjects with lateral PDOF, agreement was 89% (17/19) compared to 44% for anterior PDOF (4/9). Interobserver agreement for the Young and Tile classification schemes was moderate (weighted kappa 0.44 and 0.54, respectively). Conclusion: Crash site investigation and pelvic radiography may provide conflicting information about primary direction of injuring forces. Presumed anterior impact based on PDOF is not in consistent agreement with the pattern of injury evident on the AP pelvic radiograph.
- Published
- 2007
45. Local increase in thymic stromal lymphopoietin induces systemic alterations in B cell development
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David J. Rawlings, Theingi Aye, Miyuki Omori, Alexander Astrakhan, Masanori Iseki, Charles E. Alpers, Shirly Becker-Herman, Kelly L. Hudkins, Thuc Nghi Nguyen, Andrew G. Farr, James Dooley, and Steven F. Ziegler
- Subjects
Thymic stromal lymphopoietin ,Transgene ,medicine.medical_treatment ,Immunology ,Population ,B-Lymphocyte Subsets ,Mice, Transgenic ,Biology ,Mice ,Immune system ,Thymic Stromal Lymphopoietin ,Keratin ,medicine ,Animals ,Immunology and Allergy ,education ,B cell ,chemistry.chemical_classification ,B-Lymphocytes ,education.field_of_study ,Cell Differentiation ,Marginal zone ,Cell biology ,Cytokine ,medicine.anatomical_structure ,chemistry ,Cytokines - Abstract
The cytokine thymic stromal lymphopoietin (TSLP) drives immature B cell development in vitro and may regulate T helper type 2 responses. Here we analyzed the involvement of TSLP in B cell development in vivo with a doxycycline-inducible, keratin 5-driven transgene encoding TSLP (K5-TSLP). K5-TSLP-transgenic mice given doxycycline showed an influx of immature B cells into the periphery, with population expansion of follicular mature B cells, near-complete loss of marginal zone and marginal zone precursor B cells, and 'preferential' population expansion of peritoneal B-1b B cells. These changes promoted cryoglobulin production and immune complex-mediated renal disease. Identical events occurred in mice without T cells, in alternative TSLP-transgenic models and in K5-TSLP-transgenic mice with undetectable systemic TSLP. These observations suggest that signals mediating localized TSLP expression may modulate systemic B cell development and promote humoral autoimmunity.
- Published
- 2007
46. Wiskott-Aldrich syndrome protein is required for regulatory T cell homeostasis
- Author
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Stephanie Humblet-Baron, Steve F. Ziegler, Troy R. Torgerson, Socheath Khim, Daniel J. Campbell, Stephanie Anover, Hans D. Ochs, Shirly Becker-Herman, Blythe D. Sather, Kelly Hudkins-Loya, Charles E. Alpers, David J. Rawlings, Debora J. Kasprowicz, and Thuc Nghi Nguyen
- Subjects
Male ,Adoptive cell transfer ,Thymic stromal lymphopoietin ,Wiskott–Aldrich syndrome ,Regulatory T cell ,T cell ,Autoimmunity ,chemical and pharmacologic phenomena ,macromolecular substances ,In Vitro Techniques ,Lymphocyte Activation ,medicine.disease_cause ,T-Lymphocytes, Regulatory ,Mice ,medicine ,Animals ,Homeostasis ,Humans ,Mice, Knockout ,biology ,Wiskott–Aldrich syndrome protein ,FOXP3 ,Cell Differentiation ,Forkhead Transcription Factors ,hemic and immune systems ,General Medicine ,medicine.disease ,Adoptive Transfer ,Mice, Mutant Strains ,Wiskott-Aldrich Syndrome ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Mutation ,Immunology ,biology.protein ,Female ,Wiskott-Aldrich Syndrome Protein ,Signal Transduction ,Research Article - Abstract
Wiskott-Aldrich syndrome protein (WASp) is essential for optimal T cell activation. Patients with WAS exhibit both immunodeficiency and a marked susceptibility to systemic autoimmunity. We investigated whether alterations in Treg function might explain these paradoxical observations. While WASp-deficient (WASp(-/-)) mice exhibited normal thymic Treg generation, the competitive fitness of peripheral Tregs was severely compromised. The total percentage of forkhead box P3-positive (Foxp3(+)) Tregs among CD4(+) T cells was reduced, and WASp(-/-) Tregs were rapidly outcompeted by WASp(+) Tregs in vivo. These findings correlated with reduced expression of markers associated with self-antigen-driven peripheral Treg activation and homing to inflamed tissue. Consistent with these findings, WASp(-/-) Tregs showed a reduced ability to control aberrant T cell activation and autoimmune pathology in Foxp3(-/-)Scurfy (sf) mice. Finally, WASp(+) Tregs exhibited a marked selective advantage in vivo in a WAS patient with a spontaneous revertant mutation, indicating that altered Treg fitness likely explains the autoimmune features in human WAS.
- Published
- 2007
47. Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance
- Author
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Alexander van der Bourg, Maya Mills, Amy S. Chuong, Adrian Cheng, Andrea Benucci, Atsushi Miyawaki, Ladan Egolf, Matteo Carandini, Nathan C. Klapoetke, Fritjof Helmchen, Bosiljka Tasic, Edward S. Boyden, Susan M. Sunkin, Lu Li, R. Clay Reid, Yusuke Niino, Andras Nagy, Claudio Monetti, Ruth M. Empson, Hong Gu, Linda Madisen, Thomas Knöpfel, Thuc Nghi Nguyen, Aleena R. Garner, Hongkui Zeng, Daisuke Shimaoka, University of Zurich, Zeng, Hongkui, Massachusetts Institute of Technology. Media Laboratory, McGovern Institute for Brain Research at MIT, Chuong, Amy S, Klapoetke, Nathan Cao, and Boyden, Edward
- Subjects
Genetically modified mouse ,0303 health sciences ,10242 Brain Research Institute ,Effector ,General Neuroscience ,Transgene ,Neuroscience(all) ,2800 General Neuroscience ,Locus (genetics) ,610 Medicine & health ,Computational biology ,Optogenetics ,Biology ,3. Good health ,Viral vector ,03 medical and health sciences ,0302 clinical medicine ,Gene expression ,Recombinase ,570 Life sciences ,biology ,Neuroscience ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
available in PMC 2016 March 04, An increasingly powerful approach for studying brain circuits relies on targeting genetically encoded sensors and effectors to specific cell types. However, current approaches for this are still limited in functionality and specificity. Here we utilize several intersectional strategies to generate multiple transgenic mouse lines expressing high levels of novel genetic tools with high specificity. We developed driver and double reporter mouse lines and viral vectors using the Cre/Flp and Cre/Dre double recombinase systems and established a new, retargetable genomic locus, TIGRE, which allowed the generation of a large set of Cre/tTA-dependent reporter lines expressing fluorescent proteins, genetically encoded calcium, voltage, or glutamate indicators, and optogenetic effectors, all at substantially higher levels than before. High functionality was shown in example mouse lines for GCaMP6, YCX2.60, VSFP Butterfly 1.2, and Jaws. These novel transgenic lines greatly expand the ability to monitor and manipulate neuronal activities with increased specificity., National Institutes of Health (U.S.) (NIH grant DA028298), Wellcome Trust (London, England) (Grant), National Institutes of Health (U.S.) (NIH grant MH085500)
- Published
- 2015
48. A mesoscale connectome of the mouse brain
- Author
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Nicholas Cain, Allan R. Jones, Michael Hawrylycz, Karla E. Hirokawa, Phillip Bohn, Stefan Mihalas, Chris Lau, Eric Nicholas, Chinh Dang, Brent Winslow, Staci A. Sorensen, Seung Wook Oh, Julie A. Harris, Clifford R. Slaughterbeck, Quanxin Wang, Kevin M. Joines, Anh Ho, Wayne Wakeman, Hanchuan Peng, John G. Hohmann, Yang Li, Benjamin Ouellette, John W. Phillips, Hongkui Zeng, Lydia Ng, Paul Wohnoutka, Thuc Nghi Nguyen, Amy Bernard, Alex M. Henry, David Feng, Leonard Kuan, Christof Koch, Marty Mortrud, and Charles R. Gerfen
- Subjects
Nervous system ,Male ,Multidisciplinary ,Mesoscale meteorology ,Neuroanatomical Tract-Tracing Techniques ,Brain ,Biology ,Bioinformatics ,3d topography ,Article ,Reference space ,medicine.anatomical_structure ,medicine ,Connectome ,Biological neural network ,Animals ,Neuroscience ,Tractography - Abstract
Comprehensive knowledge of the brain's wiring diagram is fundamental for understanding how the nervous system processes information at both local and global scales. However, with the singular exception of the C. elegans microscale connectome, there are no complete connectivity data sets in other species. Here we report a brain-wide, cellular-level, mesoscale connectome for the mouse. The Allen Mouse Brain Connectivity Atlas uses enhanced green fluorescent protein (EGFP)-expressing adeno-associated viral vectors to trace axonal projections from defined regions and cell types, and high-throughput serial two-photon tomography to image the EGFP-labelled axons throughout the brain. This systematic and standardized approach allows spatial registration of individual experiments into a common three dimensional (3D) reference space, resulting in a whole-brain connectivity matrix. A computational model yields insights into connectional strength distribution, symmetry and other network properties. Virtual tractography illustrates 3D topography among interconnected regions. Cortico-thalamic pathway analysis demonstrates segregation and integration of parallel pathways. The Allen Mouse Brain Connectivity Atlas is a freely available, foundational resource for structural and functional investigations into the neural circuits that support behavioural and cognitive processes in health and disease.
- Published
- 2014
49. Synthesis of Duplex DNA Containing a Spin Labeled Analog of 2′ Deoxycytidine
- Author
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James J. Kirchner, Paul B. Hopkins, Paula L. Fischhaber, A.W. Reese, Thuc Nghi Nguyen, Bruce H. Robinson, and Eric J. Hustedt
- Subjects
Quantitative Biology::Biomolecules ,Phosphoramidite ,Dna duplex ,Stereochemistry ,Chemistry ,Biochemistry ,Chemical synthesis ,law.invention ,chemistry.chemical_compound ,law ,Genetics ,Condensed Matter::Strongly Correlated Electrons ,Deoxycytidine ,Spin label ,Electron paramagnetic resonance ,Spin labeled ,Macromolecule - Abstract
We report the chemical synthesis of phosphoramidite 8, containing a spin labeled analog of deoxycytidine, C∗, and its incorporation into synthetic DNA. The EPR characteristics of the resulting DNAs indicated that the motion of the spin label was well-correlated with the uniform modes of the macromolecule, but that correlation of the spin label with internal motion was less effective than that achieved using a spin labeled quinolone, Q.
- Published
- 1997
50. 7.4 Biophysics of Cadherin-Mediated Cell–Cell Adhesion
- Author
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Wenting Shih, A. Cheung, Soichiro Yamada, and Thuc Nghi Nguyen
- Subjects
Multicellular organism ,Cell–cell interaction ,Cadherin ,Catenin ,Biophysics ,Cell migration ,Adhesion ,Biology ,Actin cytoskeleton ,Cell adhesion ,Cell biology - Abstract
Cadherin-mediated cell–cell adhesion is a key mechanical integrator of multicellular structure and movement. The cadherin junctions are under constant mechanical load whenever cell-to-cell interactions require remodeling, a common occurrence during embryogenesis, tissue repair and cancer cell invasion. Rapid adaptation of adhesive strength is regulated by the actin cytoskeleton, yet the precise mechanisms remain ambiguous. Diverse experimental approaches including genetic, biochemical and biophysical analyses are being used to decipher the molecular interactions at the cadherin–actin linkage, which are now shaping the modern view of cadherin junctions as mechanosensing complexes.
- Published
- 2012
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