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Biology subjects

Prakash, M.

Publications and source records attributed to Prakash, M..

At least 19 recordsLinked to original sources

Diatom Logic: Rewritable active nematic circuits in tidal-flat ecosystems

At the ocean-land interface lies a critical yet largely ignored ecosystem of tidal flats[1]. These habitats are colonized by diatoms that form thin surface biofilms, creating ecosystem-scale cellular collectives stretching meters to kilometers[2-4]. How these collectives coordinate functions across vast spatiotemporal scales remains poorly understood. This task requires both long-distance transfer and long-term storage of information[5, 6]. In cellular collectives, trigger waves have been known to provide rapid, non-attenuating signaling[7-9], while extracellular matrices allow cells to externalize structural memory[10-12]. Yet, how propagating signals and structural memory integrate to shape collective functions remains elusive. Combining field observations with laboratory reconstitution, we show that tidal-flat diatoms assemble active nematic circuits that physically wire trigger-wave routing into structural memory. Here, trigger waves recruit resting diatoms into polarized motion steered by the nematic director, and topological defects act as gates that focus or bifurcate propagating wave fronts. Memory follows a fast-read, slow-write rule: rapid signals deform the cellular nematic, while slowly evolving cell-deposited mucilage retains a hidden orientational template that guides alignment recovery. Strikingly, recurrent waves progressively remodel the nematic architecture, letting the collective reshape its future routing. Coupling rapid wave propagation to extracellular memory allows collectives to resolve the stability-plasticity dilemma, maintaining structural cohesion while adapting to recurrent stimuli. Our work reveals previously hidden adaptive nematic circuits that self-organize at ecological scales, establishing a physical framework for time-programmable active materials.

biophysics↗

Survival Benefits Outweigh Germline Competition Costs in Kin Chimeras

Chimerism, the coexistence of genetically distinct cells within an organism, is a widespread phenomenon in nature. In the colonial chordate Botryllus schlosseri, fusion between compatible colonies can lead to stem cell competition with a single genotype taking over the gonadal and/or somatic tissue of the chimera. Over five years, we quantified the frequency and consequences of chimeric formation among 809 larvae, assessing the rates of fusion after natural settlement and its long-term impact on colony survival. We found larvae preferentially settle near kin, facilitating high rates of successful fusion. Fusion was concentrated around the first month of life, boosting survival during this period of high mortality. At all timepoints chimerism significantly increased survival relative to naive and rejected colonies and extended overall lifespan. However, fusion triggered intense internal competition: in all cases tested, a single genotype achieved dominance over both germline and somatic tissues within several weeks post-fusion, regardless of initial genotype contribution. We estimate that among kin, the survival benefits of chimerism outweigh the potential costs of germline loss, extending Hamiltons kin selection framework to colonial marine organisms. In this context, chimerism functions as an adaptive strategy that enhances early survival while enabling selection among competing stem cell lineages.

evolutionary biology↗

An Advanced Mobile Laboratory to enable field-based microbial ecology and cell biology across scales

Microbial biodiversity is central to ecosystem function, yet mechanistic insights into the cell biology of environmental organisms remain limited. The underlying challenges are twofold: most microbes remain uncultivable, and a persistent gap exists between field sampling and laboratory analyses. Here, we introduce the Advanced Mobile Laboratory (AML), a field-deployable platform that integrates confocal microscopy, image-enabled cell sorting, and cryo-preparation for expansion and electron microscopy. This setup enables immediate, standardized processing and analysis of environmental communities directly at the sampling site. We demonstrate its capability using marine eukaryotic plankton, showing how the AML enables multiscale investigations, from live imaging of natural communities to enabling ultrastructural and single-cell omics analyses, while minimizing sample degradation and enabling on-site experimentation. By bringing high-end sample preparation and analytical capacity into the field, the AML enables studying life in its natural context to mechanistically understand lifes diversity in the environment.

cell biology↗

Comprehensive Mapping of the Virus and Host Factors that Guide the Paths of HIV-1 Escape from a Therapeutic

HIV-1 resistance to therapeutics can emerge through diverse mutational routes, yet the determinants guiding pathway selection in vivo remain unclear. Through comprehensive screening, we identified 18 mutations in the HIV-1 Env protein that enhance resistance to the FDA-approved small-molecule therapeutic temsavir. We then examined their occurrence in HIV-infected individuals who developed resistance on therapy. Only a subset of the resistance-enhancing mutations emerged in vivo. On-treatment mutation frequencies correlated with their spontaneous emergence rates in temsavir-untreated individuals, and were governed by two parameters: (i) Probability of mutation appearance, determined by number and type of nucleotide changes required, and (ii) Probability of mutation persistence, determined by Env functional and immune fitness. Notably, non-neutralizing antibodies commonly-elicited in HIV-infected individuals restricted emergence of multiple resistant forms, driving convergence to a narrow set of escape routes. These findings establish a quantitative framework for predicting therapeutic resistance and reveal how host-immunity constrains viral evolution during treatment.

microbiology↗

Control of Synaptic Communication through Molecularly Engineered Bioluminescence Light Emission and Sensing

Synapses are sites of intercellular communication between neurons and from neurons to target organs, and of signal integration that underly physiological and behavioral responses. We have developed a modular platform, Interluminescence (Int), for experimental control of synaptic transmission: bioluminescent light, generated by a luciferase oxidizing a luciferin, from a presynaptic neuron is used to activate transsynaptic optogenetic ion channels in the postsynaptic neuron. Two strategies can modulate postsynaptic neurons in the presence of luciferin. In the Act-Int approach, a luciferase is genetically expressed in synaptic vesicles and released during depolarization-induced presynaptic vesicle fusion and exocytosis. In the Persist-Int approach, a luciferase is tethered to the presynaptic membrane where it can support sustained transsynaptic signaling. Both strategies can activate postsynaptic neurons with comparable efficacy under the conditions tested. By design, the modularity of the platform permits the use of luciferases and opsins ranging in brightness and light sensitivity, with the luciferase targeted to different subcellular regions of the presynaptic neuron, and the postsynaptic opsin being excitatory or inhibitory. Our results demonstrate the utility and versatility of Interluminescence to mediate synapse-specific transmission that is either activity-dependent or activity-independent.

neuroscience↗

Fluid-Squid: DIY Multiplexed Imaging of Cells and Tissues

Recent advances in multiplexed single-cell characterization have revolutionized our insight into cell biology, but many available technologies remain limited by high costs or a lack of customizability. To address these challenges, we developed Fluid-Squid, a cost-effective, quantitative imaging platform that integrates automated fluidics to support customizable, do-it-yourself multiplexed imaging workflows. Using Fluid-Squid, we successfully imaged fresh frozen human intestinal tissues with a 36-plex oligonucleotide-barcoded antibody panel and further demonstrated the feasibility of lyophilizing such multiplexed panels. We also adapted existing multiplexed imaging workflows to characterize individual cells to identify immune cell populations, phenotype, and antigen-specific cells from mouse splenocytes and human peripheral blood mononuclear cells (PBMCs) with a 39-antibody panel. To further expand its utility, we developed a barcoding strategy that allows for the pooling and simultaneous staining of multiple samples, reducing time, costs, and batch effects in single-cell experiments. This approach facilitated rapid titration to optimize antibody concentrations and assess the impact of various blood preparation methods on cell type retention. Overall, our work provides a new open-source framework for automated fluidics and microscopy in a flexible, cost-effective platform, empowering adaptable multiplexed characterization of both single cells and tissues.

bioengineering↗

Ballistic Microscopy (BaM)

Light and electron microscopy utilizes interactions of either photons or electrons with matter to create images from cellular to atomic scale. However, these methods are limited in de novo discovery and spatial mapping of unknown biomolecules. Label free methods such as mass spectrometry or sequencing lack live-cell and subcellular context. Here we introduce a new approach, Ballistic Microscopy (BaM), to image cells with physical nanoparticles. We bombard living cells with millions of nanoparticles traveling at [~]1000 m/s. Each particle passes through cells, piercing and capturing attoliters of cytoplasm on a hydrogel substrate while preserving spatial information (SPLAT-MAP). This "physical image" of a live cell captures a molecular fingerprint of a cell on a hydrogel film that can be processed post-capture via multiple techniques such as TEM, Cryo-EM, mass spectrometry, confocal imaging, and DNA amplification. Using BaM, we discover previously unknown composition of CLIP170 and Tau3R condensates in HEK cells, uncovering Keratin-18 as a structural element. BaM establishes a new paradigm of "physical imaging" with modular readout platform for spatially resolved live sampling across cells, tissues, and organisms.

cell biology↗

Long-range actin-driven endosymbiont mobility in a deep-diverging bilaterian

AbstractSymbiosis is everywhere, and "we have never been individuals"[1, 2]. In animal-microbe symbioses, established symbionts are often thought to be confined to a specific cellular or tissue niche[3-7] and generally lose their motile appendages such as flagella[8-15]. However, whether the loss of motile appendages necessarily implies immobility within the animal host remains an open conundrum. Here, we present the discovery of long-range, host actin-driven symbiont mobility in a dinoflagellate-acoel worm symbiosis. Using long-term tracking, fluorescence, and electron microscopy, we find that dinoflagellate symbionts (Amphidinium sp., 10-20{micro}m in size) travel throughout an extensive network of thin host cells ([~]200 nm in regions without symbionts) in Waminoa sp. acoel worms, which are part of a deep-diverging bilaterian lineage[16-18]. Although FIB-SEM-based 3D reconstruction shows symbionts still retain both flagella, we uncover that it is host actin machinery that plays a primary role in overcoming large drag forces under confinement to achieve mobility throughout the worm at surprisingly high velocities (around 1{micro}m/s). Long term in-toto imaging further reveals diel rhythms and spatiotemporal regulation of symbionts during regeneration. Our findings show the presence of host-mediated mobility in animal-microbe symbioses, which suggests the existence of previously overlooked regulatory processes in holobionts maintenance of dynamic homeostasis.

biophysics↗

Schistosomiasis parasite enhance transmission rates via interfacial swimming

Schistosomiasis is a disease of poverty that affects over 240 million people worldwide despite an ecological paradox: cercariae larvae are short-lived (12 hours), sparse in the water body, and entrained by flows exceeding their swimming speeds - conditions that should limit host-finding. We resolve this paradox by demonstrating that cercariae utilize a physics-based strategy: they actively accumulate at the air-water interface, transforming inefficient three-dimensional search into effective two-dimensional exploration, increasing host-encounter efficiency by a thousand fold. Our biophysical models demonstrate that this behavior emerges from weight-asymmetric morphology, enabling an embodied algorithm for surface-seeking swimming modes without complex neural control. Surface-swimming cercariae benefit from near-zero vertical flows near the surface, allowing them to remain there while exploiting wind-driven currents for long-distance dispersal. We identify the air-water interface as a critical transmission micro-habitat, suggesting new physical intervention strategies targeting this interface to mitigate transmission of this highly infectious disease.

biophysics↗

Tubulin isotypes polymerise into sectioned microtubules that locally regulate protein binding

Microtubules assemble from tubulin heterodimers composed of conserved - and {beta}- tubulins. In many species, including humans, tubulins are expressed from multiple genes. While the resulting tubulin isotypes show only subtle sequence differences, microtubules made of distinct isotypes can differ in their dynamic behaviour, as well as in their structure, for example in the number of their protofilaments. In cells, tubulin isotypes co-polymerize into mixed-isotype microtubules. How the mixing of tubulin isotypes affects microtubule functionality is unknown. Here we show that co-polymerization of recombinant tubulin dimers containing two different human {beta}-tubulin isotypes, 1{beta}3 and 1{beta}4, generates sectioned microtubules in which tubulin content and protofilament number differ from one section to the next. We demonstrate that two microtubule-associated proteins (MAPs), TPPP1 and optineurin (OPTN), bind differentially to these sections. Microtubules grown from natively mixed-isotype sources, either HeLa cells or mammalian brain tissue, also consist of sections that are differentially recognized by TPPP1 and OPTN, suggestive of isotype sorting. Our results demonstrate that co-polymerization of multiple tubulin isotypes can drive microtubules to assemble into distinct sections that locally regulate the interactions of MAPs with the microtubule lattice.

cell biology↗

Impact of missing data correlated with labels to be predicted in neurodegeneration classification tasks

We introduce a new subtype of Missing Not at Random (MNAR) data, where the missingness is correlated with the labels (y) to be predicted, termed (y)-dependent MNAR. We demonstrate that this subtype can significantly bias the estimation of performance metrics in typical machine learning tasks. Unbiased error estimation is crucial in predictive modeling to accurately assess model performance, identify potential biases, and ensure generalizability to new, unseen data. We explore the effects of imputing this new subtype of MNAR and compare it with general missing types, namely Missing at Random (MAR) and Missing Completely at Random (MCAR). Our comparison analysis employs both synthetic and clinical datasets, including the Alzheimers Disease Neuroimaging Initiative (ADNI) dataset, the Parkinsons Progression Markers Initiative (PPMI) dataset, and the Anti-Amyloid Treatment in Asymptomatic Alzheimers Disease (A4) dataset. After introducing missingness into the datasets, we trained different classifiers paired with various imputation methods and measured repeated cross-validation test metrics. Our findings reveal that datasets with non-ignorable missing types (MNAR) exhibit a strong bias compared to ignorable types (MAR and MCAR) in downstream analysis. Non-linear classifiers tend to exploit patterns from imputed data, particularly when the imputed values correlate with the target label (y), which can lead to unreliable estimation of the generalization error. Mean and median imputations proved to be more robust than tree-based or gradient boosting methods.

bioinformatics↗

Ice gliding diatoms establish record-low temperature limit for motility in a eukaryotic cell

Despite periods of permanent darkness and extensive ice coverage in polar environments, photosynthetic ice diatoms display a remarkable capability of living inside the ice matrix. How these organisms navigate such hostile conditions with limited light and extreme cold remains unknown. Using a custom sub-zero temperature microscope during an Arctic expedition, we present the discovery of motility at record-low temperatures in a Eukaryotic cell. By characterizing the gliding motility of several ice diatom species, collected from ice cores in the Chukchi Sea, we record that they retain motility at temperatures as low as -15 {degrees}C. Remarkably, ice diatoms can glide on ice substrates, a capability absent in temperate diatoms of the same genus. This unique ability arises from adaptations in extracellular mucilage that allow ice diatoms to adhere to ice, essential for gliding. Even on glass substrates where both cell types retain motility at freezing temperatures, ice diatoms move an order of magnitude faster, with their optimal motility shifting toward colder temperatures. Combining field and laboratory experiments with thermo-hydrodynamic modeling, we reveal adaptive strategies that enable gliding motility at extreme cold temperatures. These strategies involve increasing internal energy efficiency with minimal changes in heat capacity and activation enthalpy, and reducing external dissipation by minimizing the temperature sensitivity of mucilage viscosity. The discovery of diatoms ice gliding motility opens new routes for understanding their survival within a harsh ecological niche and their migratory responses to rapid environmental changes. Our work highlights the robust adaptability of ice diatoms in one of Earths most extreme settings.

biophysics↗

Bridging biomolecular modalities for knowledge transfer in bio-language models

In biology, messenger RNA (mRNA) plays a crucial role in gene expression and protein synthesis. Accurate predictive modeling of mRNA properties can greatly enhance our understanding and manipulation of biological processes, leading to advancements in medical and biotechnological applications. Utilizing bio-language foundation models allows for leveraging large-scale pretrained knowledge, which can significantly improve the efficiency and accuracy of these predictions. However, mRNA specific foundation models are notably limited posing challenges for efficient predictive modeling in mRNA-focused tasks. In contrast, DNA and protein modalities have numerous general-purpose foundation models trained on billions of sequences. This paper explores the potential for adaptation of existing DNA and protein bio-language models for mRNA-focused tasks. Through experiments using various mRNA datasets curated from both public domain and internal proprietary database, we demonstrate that pre-trained DNA and protein models can be effectively transferred for mRNA-focused tasks using various adaptation techniques such as probing, full-rank, and low-rank finetuning. In addition, we identify key factors that influence successful adaptation, offering guidelines on when general-purpose DNA and protein models are likely to perform well for mRNA-focused tasks. We further assess the impact of model size on adaptation efficacy, finding that medium-scale models often outperform larger ones for cross-modal knowledge transfer. We conclude that by leveraging the interconnectedness of DNA, mRNA, and proteins, as outlined by the central dogma of molecular biology, the knowledge in foundation models can be effectively transferred across modalities, significantly enhancing the repertoire of computational tools available for mRNA analysis.

bioinformatics↗

Single Cells Signal Non-self from Self via 'XOR' and 'NOT EQUALS' Logic in a Dimer of Dimers

The ciliate Tetrahymena thermophilas seven mating types are defined by unique receptor-ligand pairs (Mta/Mtb). While Mta and Mtb are known to participate in a mating signal complex, how they distinguish between oneself and six non-self cell types remains unknown. AlphaFold3 predictions reveal Mta/Mtb as large glycoproteins likely derived from ancient, unisexual, intercellular adhesion molecules. Since homologous binary-type systems perform XOR by switching mtA expression, we show spectrum (n) types naturally extend XOR to multi-bit NOT EQUALS operations via differential affinities of Mta/Mtb dimers. We model kinetics begetting the n + 1th type, demonstrating self-inhibition by trans-homophilic Mtb-Mtb. A computational approach reconciles recent and classical evidence for mating exclusivity, including selfing failures (same type mating). Binding kinetics enables fast, robust intercellular computation across an intermembrane mating space. Thus, Mta/Mtb families are a model system allowing us to derive a calculus of antigenic variation and inspire synthetic designs of XOR logic underlying self/non-self recognition. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=198 HEIGHT=200 SRC="FIGDIR/small/616003v1_ufig1.gif" ALT="Figure 1"> View larger version (46K): org.highwire.dtl.DTLVardef@572f8forg.highwire.dtl.DTLVardef@1f386b2org.highwire.dtl.DTLVardef@1104fb1org.highwire.dtl.DTLVardef@1e006a1_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIWithin minutes, swimming cells signal compatible mating types via membrane proteins C_LIO_LIAlphaFold insights reveal Mta/Mtb family as elongate adhesion receptors/ligands C_LIO_LIRelative affinities of two dimers explain multi-bit intercellular signaling capacity C_LIO_LIDimer-of-dimers compute NOT EQUALS via homo-vs. hetero-philic interactions C_LIO_LIIn Brief Free-living ciliates T. thermophila breed only when two cells are different in mating type, out of seven types total. Their mating-type-specific coexpression of a receptor-ligand glycoprotein pair reveals how competitive homophilic vs. heterophilic binding computes self vs. non-self recognition with single-layer intercellular logic. Proteins perform binary XOR gates (with two mating types) that extend to multi-bit NOT EQUALS (with multiple mating types). The work illuminates principles of eXclusivity in molecular recognition and inspires the design of synthetic cellular recognition systems. C_LI

systems biology↗

Creased ciliary flocks shape unfolding dynamics via information bottlenecks in an aneural animal

Multicellular organisms utilize thin sheet folding to achieve functional three-dimensional forms. During embryonic development, stereotypical epithelial folds emerge from active cellular and molecular processes including cell shape change and differential cell growth. Active thin sheet folding promises to be a powerful design technique in the fields of active solids, soft robotics, and synthetic biology. However, the general principles of active thin sheet folding remain poorly understood. Here we discover a non-canonical cilia-driven thin sheet folding behavior exhibited by basal animal Trichoplax adhaerens. Through volumetric imaging, we found that, despite having no nervous system, T. adhaerens has the remarkable ability to resolve complex body folding states in a non-stereotypical fashion using a carpet of collectively flocking cilia. Cilia-resolved imaging revealed that folds create crease defects in the animals ciliary carpet, which act as information bottlenecks to break collective behavior of cilia. In turn, these bottlenecks enable the disjointed locomotion required for fold removal. These findings point to a two-way coupling mechanism, wherein ciliary activity shapes the animals folding state and vice versa. Our work demonstrates the broad configuration space of non-stereotypical active folding and highlights the power of distributed activity to drive folding and unfolding of a thin multicellular sheet. We anticipate our study to be a starting point for the establishment of a new class of distributed active origami wherein fold lines themselves are dynamic and motile, with implications in engineering of self-folding materials. Additionally, our work reveals a new facet of the Placozoan behavioral repertoire, which extends our understanding of mechanical intelligence in the absence of a nervous system.

biophysics↗

Biophysical limits of ultrafast cellular motility

Many single-celled organisms and specialized cell types can surprisingly achieve speed and acceleration significantly faster than multicellular counterparts. These remarkable cellular machines must integrate energy storage and amplification in actuation, latches for triggered release, and energy dissipation without failure -- all implemented in macro-molecular assemblies inside a single cell. However, a universal biophysical framework that can comparatively evaluate extreme cellular motility remains lacking. Scaling laws have long been recognized as powerful tools for revealing universal principles in physical systems. We map the atlas of ultrafast motility for single cells across the tree of life. We then introduce a new quantitative framework that can be used to evaluate and compare extreme acceleration, speed, area strain rate, volume expansion strain rate, and density changes in single cells. Recognizing that many single cells operate in low-Reynolds number environments, we introduce a new dimensionless number, the "cellular acceleration number," based on energy dissipation at this scale. Using this new framework, we discover a scaling law between the cellular acceleration number and the transient Reynolds number, valid across six orders of magnitude in a range of single-cell organisms. We further generalize these ideas by placing various trigger, actuation, and dissipation mechanisms within the same framework and estimating the fundamental limits of speed and acceleration at the cellular scale. We conclude with a detailed summary of the range of functions implemented via ultrafast cellular phenomena, laying down a quantitative foundation for extreme biophysics at the cellular scale.

biophysics↗

Flight-Scope: microscopy with microfluidics in microgravity

With the European Space Agency (ESA) and NASA working to return humans to the moon and onwards to Mars, it has never been more important to study the impact of altered gravity conditions on biological organisms. These include astronauts but also useful micro-organisms they may bring with them to produce food, medicine, and other useful compounds by synthetic biology. Parabolic flights are one of the most accessible microgravity research platforms but present their own challenges: relatively short periods of altered gravity ([~]20s) and aircraft vibration. Live-imaging is necessary in these altered-gravity conditions to readout any real-time phenotypes. Here we present Flight-Scope, a new microscopy and microfluidics platform to study dynamic cellular processes during the short, altered gravity periods on parabolic flights. We demonstrated Flight-Scopes capability by performing live and dynamic imaging of fluorescent glucose uptake by yeast, S. cerevisiae, on board an ESA parabolic flight. Flight-Scope operated well in this challenging environment, opening the way for future microgravity experiments on biological organisms.

biophysics↗

A genetic and microscopy toolkit for live imaging of whole-body regeneration in Macrostomum lignano

Live imaging of regenerative processes can reveal how animals restore their bodies after injury through a cascade of dynamic cellular events. Here, we present a comprehensive toolkit for live imaging of whole-body regeneration in the flatworm Macrostomum lignano, including a high throughput cloning pipeline, targeted cellular ablation, and advanced microscopy solutions. Using tissue-specific reporter expression, we examine how various structures regenerate. Enabled by a custom, low-cost luminescence/fluorescence microscope, we overcome intense stress-induced autofluorescence to demonstrate the first application of genetic cellular ablation in flatworms to reveal the limited regenerative capacity of neurons and their essential role during wound-healing. Finally, we build a novel open-source tracking microscope to continuously image moving animals throughout the week-long process of regeneration, quantifying kinetics of wound healing, nerve cord repair, body regeneration, growth, and behavioral recovery. Our findings suggest that nerve cord reconnection operates independently from primary body axis re-establishment and other downstream regenerative processes.

bioengineering↗