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Mitra, P. P.

Publications and source records attributed to Mitra, P. P..

At least 19 recordsLinked to original sources

Discrete Morse Graph Construction for High-Dimensional Transcriptomic Data

Single-cell transcriptomic atlases resolve thousands of putative brain cell types, yet common analytical workflows often rely on low-dimensional embeddings that can distort neighborhood relationships and obscure continuous variation between closely related populations. We introduce single-cell Discrete Morse Graph Construction (scDMGC), a deterministic framework based on topological data analysis and discrete Morse theory that extracts a compact graph from neighborhood structure in high-dimensional gene-expression space. Because graph edges are derived from the original k-nearest-neighbor complex, scDMGC provides an interpretable multiscale representation of transcriptomic organization without requiring a low-dimensional embedding. Local maxima represent coherent transcriptional states, saddles quantify their separation, and gradient paths capture intermediate states and continuous transitions. Applied to cortical and hippocampal datasets, scDMGC recovers established inhibitory-neuron classes and elucidates transcriptional gradients. In mouse whole-cortex data, Morse graph structure evaluates cell-type hierarchy and type robustness. Persistence across scales provides a quantitative measure of cell-type identity and discrete versus continuous relationships. Finally, in Alzheimers disease single-nucleus data, scDMGC distinguishes changes in cell-type abundance from disease-associated shifts in transcriptional state. scDMGC therefore provides a multiscale framework for quantifying whether transcriptional populations form persistent cell types, how strongly external annotations are supported by intrinsic data structure, how cell types relate to one another, and where discrete organization transitions into continuous variation.

neuroscience↗

Skeletonization of neuronal processes using Discrete Morse techniques from computational topology

To understand biological intelligence we need to map neuronal networks in vertebrate brains. Mapping mesoscale neural circuitry is done using injections of tracers that label groups of neurons whose axons project to different brain regions. Since many neurons are labeled, it is difficult to follow individual axons. Previous approaches have instead quantified the regional projections using the total label intensity within a region. However, such a quantification is not biologically meaningful. We propose a new approach better connected to the underlying neurons by skeletonizing labeled axon fragments and then estimating a volumetric length density. Our approach uses a combination of deep nets and the Discrete Morse (DM) technique from computational topology. This technique takes into account nonlocal connectivity information and therefore provides noise-robustness. We demonstrate the utility and scalability of the approach on whole-brain tracer injected data. We also define and illustrate an information theoretic measure that quantifies the additional information obtained, compared to the skeletonized tracer injection fragments, when individual axon morphologies are available. Our approach is the first application of the DM technique to computational neuroanatomy. It can help bridge between single-axon skeletons and tracer injections, two important data types in mapping neural networks in vertebrates.

neuroscience↗

3D multimodal histological atlas and coordinate framework for the mouse brain and head

Brain reference atlases are essential for neuroscience experiments and data integration. However, histological atlases of the mouse brain, crucial in biomedical research, have not kept pace. Autofluorescence-based volumetric brain atlases are increasingly used but lack microscopic histological contrast, cytoarchitectonic information, corresponding MRI datasets, and often have truncated brainstems. Here, we present a multimodal, multiscale atlas of the laboratory mouse brain and head. The new reference brains include the whole head with consecutive Nissl and myelin serial section histology in three planes of section with 0.46 {micro}m in-plane resolution, including intact brainstem, cranial nerves, and associated sensors and musculature. We provide reassembled histological volumes with 20mu isotropic resolution in stereotactic coordinates, determined using co-registered in vivo MRI and CT. In addition to conventional MRI contrasts, we provide diffusion MRI-based in vivo and ex vivo microstructural information, adding a valuable co-registered contrast modality that bridges MRI with cell-resolution histological data. We shift emphasis from compartmental annotations to stereotactic coordinates in the reference brains, offering a basis for evolving annotations over time and resolving conflicting neuroanatomical judgments by different experts. This new reference atlas facilitates integration of molecular cell type data and regional connectivity, serves as a model for similar atlases in other species, and sets a precedent for preserving extra-cranial nervous system structures.

neuroscience↗

A three-dimensional histological cell atlas of the developing human brain

The human brain is believed to contain a full complement of neurons by the time of birth together with a substantial amount of the connectivity architecture, even though a significant amount of growth occurs postnatally. The developmental process leading to this outcome is not well understood in humans in comparison with model organisms. Previous magnetic resonance imaging (MRI) studies give three-dimensional coverage but not cellular resolution. In contrast, sparsely sampled histological or spatial omics analyses have provided cellular resolution but not dense whole brain coverage. To address the unmet need to provide a quantitative spatiotemporal map of developing human brain at cellular resolution, we leveraged tape-transfer assisted serial section histology to obtain contiguous histological series and unbiased imaging with dense coverage. Interleaved 20 thick Nissl and H&E series and MRI volumes are co-registered into multimodal reference volumes with 60 isotropic resolution, together with atlas annotations and a stereotactic coordinate system based on skull landmarks. The histological atlas volumes have significantly more contrast and texture than the MRI volumes. We computationally detect cells brain-wide to obtain quantitative characterization of the cytoarchitecture of the developing brain at 13-14 and 20-21 gestational weeks, providing the first comprehensive regional cell counts and characterizing the differential growth of the different brain compartments. Morphological characteristics permit segmentation of cell types from histology. We detected and quantified brain-wide distribution of mitotic figures representing dividing cells, providing an unprecedented spatiotemporal atlas of proliferative dynamics in the developing human brain. Further, we characterized the abundance and distribution of Cajal-Retzius cells, a transient cell population that plays essential roles in organizing glutamatergic cortical neurons into layers. Together, our study provides an unprecedented quantitative window into the developing human brain and the reference volumes and coordinate space should be useful for integrating spatial omics data sets with dense histological context.

neuroscience↗

Establishing neuroanatomical correspondences across mouse and marmoset brain structures

Interest in the common marmoset is growing due to evolutionarily proximity to humans compared to laboratory mice, necessitating a comparison of mouse and marmoset brain architectures, including connectivity and cell type distributions. Creating an actionable comparative platform is challenging since these brains have distinct spatial organizations and expert neuroanatomists disagree. We propose a general theoretical framework to relate named atlas compartments across taxa and use it to establish a detailed correspondence between marmoset and mice brains. Contrary to conventional wisdom that brain structures may be easier to relate at higher levels of the atlas hierarchy, we find that finer parcellations at the leaf levels offer greater reconcilability despite naming discrepancies. Utilizing existing atlases and associated literature, we created a list of leaf-level structures for both species and establish five types of correspondence between them. One-to-one relations were found between 43% of the structures in mouse and 47% in marmoset, whereas 25% of mouse and 10% of marmoset structures were not relatable. The remaining structures show a set of more complex mappings which we quantify. Implementing this correspondence with volumetric atlases of the two species, we make available a computational tool for querying and visualizing relationships between the corresponding brains. Our findings provide a foundation for computational comparative analyses of mesoscale connectivity and cell type distributions in the laboratory mouse and the common marmoset.

neuroscience↗

AI without networks

Contemporary Artificial Intelligence (AI) stands on two legs: large training data corpora and many-parameter artificial neural networks (ANNs). The data corpora are needed to represent the complexity and heterogeneity of the world. The role of the networks is less transparent due to the obscure dependence of the network parameters and outputs on the training data and inputs. This raises problems, ranging from technical-scientific to legal-ethical. We hypothesize that a transparent approach to machine learning is possible without using networks at all. By generalizing a parameter-free, statistically consistent data interpolation method, which we analyze theoretically in detail, we develop a network-free framework for AI incorporating generative modeling. We demonstrate this framework with examples from three different disciplines - ethology, control theory, and mathematics. Our generative Hilbert framework applied to the trajectories of small groups of swimming fish outperformed state-of-the-art traditional mathematical behavioral models and current ANN-based models. We demonstrate pure data interpolation based control by stabilizing an inverted pendulum and a driven logistic map around unstable fixed points. Finally, we present a mathematical application by predicting zeros of the Riemann Zeta function, achieving comparable performance as a transformer network. We do not suggest that the proposed framework will always outperform networks as over-parameterized networks can interpolate. However, our framework is theoretically sound, transparent, deterministic, and parameter free: remarkably, it does not require any compute-expensive training, does not involve optimization, has no model selection, and is easily reproduced and ported. We also propose an easily computed method of credit assignment based on this framework, to help address ethical-legal challenges raised by generative AI.

neuroscience↗

The cell type composition of the adult mouse brain revealed by single cell and spatial genomics

The function of the mammalian brain relies upon the specification and spatial positioning of diversely specialized cell types. Yet, the molecular identities of the cell types, and their positions within individual anatomical structures, remain incompletely known. To construct a comprehensive atlas of cell types in each brain structure, we paired high-throughput single-nucleus RNA-seq with Slide-seq-a recently developed spatial transcriptomics method with near-cellular resolution-across the entire mouse brain. Integration of these datasets revealed the cell type composition of each neuroanatomical structure. Cell type diversity was found to be remarkably high in the midbrain, hindbrain, and hypothalamus, with most clusters requiring a combination of at least three discrete gene expression markers to uniquely define them. Using these data, we developed a framework for genetically accessing each cell type, comprehensively characterized neuropeptide and neurotransmitter signaling, elucidated region-specific specializations in activity-regulated gene expression, and ascertained the heritability enrichment of neurological and psychiatric phenotypes. These data, available as an online resource (BrainCellData.org) should find diverse applications across neuroscience, including the construction of new genetic tools, and the prioritization of specific cell types and circuits in the study of brain diseases.

neuroscience↗

Transcriptomic cytoarchitecture reveals principles of human neocortex organization

Variation in cortical cytoarchitecture is the basis for histology-based definition of cortical areas, such as Brodmann areas. Single cell transcriptomics enables higher-resolution characterization of cell types in human cortex, which we used to revisit the idea of the canonical cortical microcircuit and to understand functional areal specialization. Deeply sampled single nucleus RNA-sequencing of eight cortical areas spanning cortical structural variation showed highly consistent cellular makeup for 24 coarse cell subclasses. However, proportions of excitatory neuron subclasses varied strikingly, reflecting differences in intra- and extracortical connectivity across primary sensorimotor and association cortices. Astrocytes and oligodendrocytes also showed differences in laminar organization across areas. Primary visual cortex showed dramatically different organization, including major differences in the ratios of excitatory to inhibitory neurons, expansion of layer 4 excitatory neuron types and specialized inhibitory neurons. Finally, gene expression variation in conserved neuron subclasses predicts differences in synaptic function across areas. Together these results provide a refined cellular and molecular characterization of human cortical cytoarchitecture that reflects functional connectivity and predicts areal specialization.

neuroscience↗

The BRAIN Initiative Cell Census Data Ecosystem: A User's Guide

Characterizing cellular diversity at different levels of biological organization across data modalities is a prerequisite to understanding the function of cell types in the brain. Classification of neurons is also required to manipulate cell types in controlled ways, and to understand their variation and vulnerability in brain disorders. The BRAIN Initiative Cell Census Network (BICCN) is an integrated network of data generating centers, data archives and data standards developers, with the goal of systematic multimodal brain cell type profiling and characterization. Emphasis of the BICCN is on the whole mouse brain and demonstration of prototypes for human and non-human primate (NHP) brains. Here, we provide a guide to the cellular and spatial approaches employed, and to accessing and using the BICCN data and its extensive resources, including the BRAIN Cell Data Center (BCDC) which serves to manage and integrate data across the ecosystem. We illustrate the power of the BICCN data ecosystem through vignettes highlighting several BICCN analysis and visualization tools. Finally, we present emerging standards that have been developed or adopted by the BICCN toward FAIR (Wilkinson et al. 2016a) neuroscience. The combined BICCN ecosystem provides a comprehensive resource for the exploration and analysis of cell types in the brain.

bioinformatics↗

Cortical glutamatergic projection neuron types contribute to distinct functional subnetworks

The cellular basis of cerebral cortex functional architecture remains not well understood. A major challenge is to monitor and decipher neural network dynamics across broad cortical areas yet with projection neuron (PN)-type resolution in real time during behavior. Combining genetic targeting and wide-field imaging, we monitored activity dynamics of subcortical-projecting (PTFezf2) and intratelencephalic-projecting (ITPlxnD1) types across dorsal cortex of mice during different brain states and behaviors. ITPlxnD1 and PTFezf2 neurons showed distinct activation patterns during wakeful resting, spontaneous movements, and upon sensory stimulation. Distinct ITPlxnD1 and PTFezf2 subnetworks were dynamically tuned to different sensorimotor components of a naturalistic feeding behavior, and optogenetic inhibition of ITsPlxnD1 and PTsFezf2 in subnetwork nodes disrupted distinct components of this behavior. Lastly, ITPlxnD1 and PTFezf2 projection patterns are consistent with their subnetwork activation patterns. Our results show that, in addition to the concept of columnar organization, dynamic areal and PN type-specific subnetworks are a key feature of cortical functional architecture linking microcircuit components with global brain networks.

neuroscience↗

Genetic dissection of glutamatergic neuron subpopulations and developmental trajectories in the cerebral cortex

Diverse types of glutamatergic pyramidal neurons (PyNs) mediate the myriad processing streams and output channels of the cerebral cortex, yet all derive from neural progenitors of the embryonic dorsal telencephalon. Here, we establish genetic strategies and tools for dissecting and fate mapping PyN subpopulations based on their developmental and molecular programs. We leverage key transcription factors and effector genes to systematically target the temporal patterning programs in progenitors and differentiation programs in postmitotic neurons. We generated over a dozen temporally inducible mouse Cre and Flp knock-in driver lines to enable combinatorial targeting of major progenitor types and projection classes. Intersectional converter lines confer viral access to specific subsets defined by developmental origin, marker expression, anatomical location and projection targets. These strategies establish an experimental framework for understanding the hierarchical organization and developmental trajectory of PyN subpopulations that assemble cortical processing networks and output channels.

neuroscience↗

Solving the where problem in neuroanatomy: a generative framework with learned mappings to register multimodal, incomplete data into a reference brain

A current focus of research in neuroscience is to enumerate, map and annotate neuronal cell types in whole vertebrate brains using different modalities of data acquisition. Mapping these molecular and anatomical datasets into a common reference space remains a key challenge. While several brain-to-atlas mapping workflows exist, they do not adequately address challenges of modern high throughput neuroimaging, including multimodal and multiscale signals, missing data or non reference signals, and geometric quantification of individual variation. Our solution is to implement a generative statistical model that describes the likelihood of imaging data given a sequence of transforms of an atlas image, and a framework for maximum a posteriori estimation of unknown parameters capturing the issues listed above. The key idea in our approach is to minimize the difference between synthetic image volumes and real data over these parameter. Rather than merely using mappings as a "normalization" step, we implement tools for using their local metric changes as an opportunity for geometric quantification of technical and biological sources of variation in an unprecedented manner. While the framework is used to compute pairwise mappings, our approach particularly allows for easy compositions across chains of multimodality datasets. We apply these methods across a broad range of datasets including various combinations of in-vivo and ex-vivo MRI, 3D STP and fMOST data sets, 2D serial histology sections, and brains processed for snRNAseq with tissue partially removed. We show biological utility by quantifying cell density and diffeomorphic characterization of brain shape fluctuations across biological covariates. We note that the magnitude of individual variation is often greater than differences between different sample preparation techniques. To facilitate community accessibility, we implement our algorithm as open source, include a web based framework, and implement input and output dataset standards. Our work establishes a quantitative, scalable and streamlined workflow for unifying a broad spectrum of multi-modal whole-brain light microscopic data volumes into a coordinate-based atlas framework. This work enables large scale integration of whole brain data sets that are essential in modern neuroscience.

neuroscience↗

Detection and skeletonization of single neurons and tracer injections using topological methods

Neuroscientific data analysis has traditionally relied on linear algebra and stochastic process theory. However, the tree-like shapes of neurons cannot be described easily as points in a vector space (the subtraction of two neuronal shapes is not a meaningful operation), and methods from computational topology are better suited to their analysis. Here we introduce methods from Discrete Morse (DM) Theory to extract the tree-skeletons of individual neurons from volumetric brain image data, and to summarize collections of neurons labelled by tracer injections. Since individual neurons are topologically trees, it is sensible to summarize the collection of neurons using a consensus tree-shape that provides a richer information summary than the traditional regional connectivity matrix approach. The conceptually elegant DM approach lacks hand-tuned parameters and captures global properties of the data as opposed to previous approaches which are inherently local. For individual skeletonization of sparsely labelled neurons we obtain substantial performance gains over state-of-the-art non-topological methods (over 10% improvements in precision and faster proofreading). The consensus-tree summary of tracer injections incorporates the regional connectivity matrix information, but in addition captures the collective collateral branching patterns of the set of neurons connected to the injection site, and provides a bridge between single-neuron morphology and tracer-injection data.

neuroscience↗

Semantic segmentation of microscopic neuroanatomical data by combining topological priors with encoder-decoder deep networks

Understanding of neuronal circuitry at cellular resolution within the brain has relied on tract tracing methods which involve careful observation and interpretation by experienced neuroscientists. With recent developments in imaging and digitization, this approach is no longer feasible with the large scale (terabyte to petabyte range) images. Machine learning based techniques, using deep networks, provide an efficient alternative to the problem. However, these methods rely on very large volumes of annotated images for training and have error rates that are too high for scientific data analysis, and thus requires a significant volume of human-in-the-loop proofreading. Here we introduce a hybrid architecture combining prior structure in the form of topological data analysis methods, based on discrete Morse theory, with the best-in-class deep-net architectures for the neuronal connectivity analysis. We show significant performance gains using our hybrid architecture on detection of topological structure (e.g. connectivity of neuronal processes and local intensity maxima on axons corresponding to synaptic swellings) with precision/recall close to 90% compared with human observers. We have adapted our architecture to a high performance pipeline capable of semantic segmentation of light microscopic whole-brain image data into a hierarchy of neuronal compartments. We expect that the hybrid architecture incorporating discrete Morse techniques into deep nets will generalize to other data domains.

neuroscience↗

Unidirectional monosynaptic connections from auditory areas to the primary visual cortex in the marmoset monkey

Until the late 20th Century, it was believed that different sensory modalities were processed by largely independent pathways in the primate cortex, with cross-modal integration only occurring in specialized polysensory areas. This model was challenged by the finding that the peripheral representation of the primary visual cortex (V1) receives monosynaptic connections from areas of the auditory cortex in the macaque. However, auditory projections to V1 have not been reported in other primates. We investigated the existence of direct interconnections between V1 and auditory areas in the marmoset, a New World monkey. Labelled neurons in auditory cortex were observed following 4 out of 10 retrograde tracer injections involving V1. These projections to V1 originated in the caudal subdivisions of auditory cortex (primary auditory cortex, caudal belt and parabelt areas), and targeted parts of V1 that represent parafoveal and peripheral vision. Injections near the representation of the vertical meridian of the visual field labelled few or no cells in auditory cortex. We also placed 8 retrograde tracer injections involving core, belt and parabelt auditory areas, none of which revealed direct projections from V1. These results confirm the existence of a direct, nonreciprocal projection from auditory areas to V1 in a different primate species, which has evolved separately from the macaque for over 30 million years. The essential similarity of these observations between marmoset and macaque indicate that early-stage audiovisual integration is a shared characteristic of primate sensory processing.

neuroscience↗

A High-throughput Neurohistological Pipeline for Brain-Wide Mesoscale Connectivity Mapping of the Common Marmoset

Understanding the connectivity architecture of entire vertebrate brains is a fundamental but difficult task. MRI based methods offer whole brain coverage, but remain indirect in the approach to connectivity mapping. Recent progress has been made in directly mapping whole-brain connectivity architecture in the mouse at the mesoscopic scale. The basic approach uses tracer injections systematically placed on a grid of locations spanning the brain and computational analysis of the resulting whole brain data sets. Scaling this approach to bigger primate brains poses nontrivial technical challenges. Here we present an integrated neurohistological pipeline as well as a grid-based tracer injection strategy for systematic mesoscale connectivity mapping in the common Marmoset (Callithrix jacchus). Individual brains are sectioned into [~]1700 20{micro}m sections using the tape transfer technique, permitting high quality 3D reconstruction of a series of histochemical stains (Nissl, myelin) interleaved with tracer labelled sections. Combining the resulting 3D volumes, containing informative cytoarchitectonic markers, with in-vivo and ex-vivo MRI, and using an integrated computational pipeline, we are able to overcome the significant individual variation exhibited by Marmosets to obtain routine and high quality maps to a common atlas framework. This will facilitate the systematic assembly of a mesoscale connectivity matrix together with unprecedented 3D reconstructions of brain-wide projection patterns in a primate brain. While component instruments or protocols may be available from previous work, we believe that this is the first detailed systems-level presentation of the methodology required for high-throughput neuroanatomy in a model primate.

neuroscience↗

Continuity between koniocellular layers of dorsal lateral geniculate and inferior pulvinar nuclei in common marmosets

In primates, the koniocellular (K) layers of the dorsal lateral geniculate nucleus (LGN) and the calbindin-rich subdivisions of the inferior pulvinar (IPul) nucleus are considered part of a thalamic matrix system which projects diffusely to superficial cortical layers. Activity in the matrix system is proposed to coordinate oscillatory activity in thalamocortical loops. Further, since both K cells and IPul are involved in visual processing pathways, especially in alternative pathways to visual cortex after V1 lesion in early life (\"blindsight\"), their functional similarities have been strongly implicated. Here we tested the hypothesis that calbindin-positive K cells and IPul cells constitute a continuous group of cells. By combining immunohistochemistry and a high-throughput neuronal tracing method, we found that both K cells and IPul form reciprocal connections with striate and extrastriate cortices; whereas principal laminae of LGN do not receive inputs from extrastriate cortex and only project sparsely to these areas. Retrograde labelled cells in lateral division of IPul merged seamlessly into the retrograde labelled cells in K layers. These results supported the continuity between LGN K layers and IPul, providing the anatomical basis for functional congruity of this part of dorsal thalamic matrix.

neuroscience↗

On variational solutions for whole brain serial-section histology using the computational anatomy random orbit model

This paper presents a variational framework for dense diffeomorphic atlas-mapping onto high-throughput histology stacks at the 20 m meso-scale. The observed sections are modelled as Gaussian random fields conditioned on a sequence of unknown section by section rigid motions and unknown diffeomorphic transformation of a three-dimensional atlas. To regularize over the high-dimensionality of our parameter space (which is a product space of the rigid motion dimensions and the diffeomorphism dimensions), the histology stacks are modelled as arising from a first order Sobolev space smoothness prior. We show that the joint maximum a-posteriori, penalized-likelihood estimator of our high dimensional parameter space emerges as a joint optimization interleaving rigid motion estimation for histology restacking and large deformation diffeomorphic metric mapping to atlas coordinates. We show that joint optimization in this parameter space solves the classical curvature non-identifiability of the histology stacking problem. The algorithms are demonstrated on a collection of whole-brain histological image stacks from the Mouse Brain Architecture Project.\n\nAuthor SummaryNew developments in neural tracing techniques have motivated the widespread use of histology as a modality for exploring the circuitry of the brain. Automated mapping of pre-labeled atlases onto modern large datasets of histological imagery is a critical step for elucidating the brains neural circuitry and shape. This task is challenging as histological sections are imaged independently and the reconstruction of the unsectioned volume is nontrivial. Typically, neuroanatomists use reference volumes of the same subject (e.g. MRI) to guide reconstruction. However, obtaining reference imagery is often non-standard, as in high-throughput animal models like mouse histology. Others have proposed using anatomical atlases as guides, but have not accounted for the intrinsic nonlinear shape difference from atlas to subject. Our method addresses these limitations by jointly optimizing reconstruction informed by an atlas simultaneously with the nonlinear change of coordinates that encapsulates anatomical variation. This accounts for intrinsic shape differences and enables rigorous, direct comparisons of atlas and subject coordinates. Using simulations, we demonstrate that our method recovers the reconstruction parameters more accurately than atlas-free models and innately produces accurate segmentations from simultaneous atlas mapping. We also demonstrate our method on the Mouse Brain Architecture dataset, successfully mapping and reconstructing over 500 brains.

neuroscience↗