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Laplante, C.

Publications and source records attributed to Laplante, C..

5 recordsLinked to original sources

SIMBA: an agentic AI platform for single-molecule multi-dimensional imaging

Advances in multi-dimensional imaging method and probe developments have brought super-resolution fluorescence microscopy into a functional era. They capture additional single-molecule fluorescence information concurrently with spatial localization, enabling simultaneous identification of molecular species and interrogation of nanoscale environments with rich, high-dimensional imaging information. However, the adoption of multi-dimensional imaging has been hindered by fragmented analysis workflows, complex parameter tuning, and limited integration of advanced computational methods. Here, we introduce an agentic single-molecule multi-dimensional bioimaging AI, referred to as SIMBA, an AI-driven platform that unifies single-molecule localization, spectral processing and deep learning-based denoising within a single agentic and interactive framework. SIMBA incorporates large language model-based agents capable of interpreting user intent, orchestrating analysis pipelines, and dynamically selecting computational tools for automated data processing. We demonstrate that SIMBA enables supports standard single-molecule localization workflow, functional mapping of nanoscale environmental heterogeneity through single-molecule spectral analysis and denoising using developed supervised learning methods. By integrating extensible tool architectures with human language-guided workflows, SIMBA establishes a new paradigm for intelligent microscopy analysis, lowering barriers to multi-dimensional imaging adoption while enabling scalable, reproducible, and adaptive analysis of complex imaging datasets.

bioengineering↗

Actin-microtubule crosstalk imparts stiffness to the contractile ring in fission yeast.

Actin-microtubule interactions are critical for cell division yet how these networks of polymers mutually influence their mechanical properties and functions in live cells remains unknown. In fission yeast, the post-anaphase array (PAA) of microtubules assembles in the plane of the contractile ring and its assembly relies on the Myp2p-dependent recruitment of Mto1p, a component of equatorial microtubule organizing centers (eMTOCs). The general organization of this array of microtubule and the impact on their physical attachment to the contractile ring remain unclear. We found that Myp2p facilitates the recruitment of Mto1p to the inner face of the contractile ring where the eMTOCs polymerize microtubules without their direct interaction. The PAA microtubules form a dynamic polygon of Ase1p crosslinked microtubules inside the contractile ring. The specific loss of PAA microtubules affects the mechanical properties of the contractile ring of actin by lowering its stiffness. This change in the mechanical properties of the ring has no measurable impact on cytokinesis or on the anchoring of the ring. Our work proposes that the PAA microtubules exploit the contractile ring for their assembly and function during cell division while the contractile ring may receive no benefit from these interactions.

cell biology↗

Microtubule competition and cell growth recenter the nucleus after anaphase in fission yeast.

Cells actively position their nucleus based on their activity. In fission yeast, microtubule-dependent nuclear centering is critical for symmetrical cell division. After spindle disassembly at the end of anaphase, the nucleus recenters over a ~90 min period, approximately half of the duration of the cell cycle. Live cell and simulation experiments support the cooperation of two distinct mechanisms in the slow recentering of the nucleus. First, a push-push mechanism acts from spindle disassembly to septation and involves the opposing actions of the mitotic Spindle Pole Body microtubules that push the nucleus away from the ends of the cell while post-anaphase array of microtubules basket the nucleus and limit its migration toward the division plane. Second, a slow-and-grow mechanism finalizes nuclear centering in the newborn cell. In this mechanism, microtubule competition stalls the nucleus while asymmetric cell growth slowly centers it. Our work underlines how intrinsic properties of microtubules differently impact nuclear positioning according to microtubule network organization and cell size.

cell biology↗

Cooperation between Imp2p and Cdc15p imparts stiffness to the constricting contractile ring in fission yeast.

The contractile ring must anchor to the plasma membrane and cell wall to transmit its tension to the plasma membrane. F-BAR domain containing proteins including Cdc15p and Imp2p in fission yeast are likely candidate anchoring proteins based on their mutant phenotypes. Cdc15p is a node component, links the actin bundle to the plasma membrane, recruits Bgs1p to the division plane, prevents contractile ring sliding, and contributes to the stiffness of the contractile ring. Less is known about Imp2p. We found that similarly to Cdc15p, Imp2p contributes to the stiffness of the contractile ring and assembles into protein clusters. Imp2p clusters contain [~]8 Imp2p dimers and depend on the actin network for their stability at the division plane. Importantly, Imp2p and Cdc15p reciprocally affect the amount of the other in the contractile ring indicating that the two proteins influence each other during cytokinesis and may explain their similar phenotypes.

cell biology↗

Genetic transformation and live-cell nuclear and actin dynamics during the life cycle of a chytrid

Chytrids are early-diverging fungi that share ancestral features of animals, including cells that crawl and swim. At later stages, chytrid cells resemble fungi with a chitin-based cell wall and hyphal-like structures known as rhizoids. Chytrids are important evolutionary transitional forms, but much remains unknown about their cell biology because we lack genetic tools for the live-cell imaging of their nuclear and cytoskeletal dynamics. Here, we generated stable transgenic lines of the soil chytrid Spizellomyces punctatus, and coupled live-cell microscopy and fluorescent tagging to measure the timing and coordination of growth, the cell cycle, and the actin cytoskeleton. We show that Spizellomyces zoospores rapidly encyst, develop rhizoids, and undergo multiple rounds of synchronous nuclear division in a sporangium, followed by cellularization, to create and release hundreds of zoospores. The life cycle is complete in less than 30 hours. We further demonstrate that crawling zoospores, akin to animal cells, display polymerized actin at the leading edge of amoeboid fronts. After encystment, polymerized actin reorganizes into fungal-like cortical patches and cables that extend into the rhizoid. Actin remains highly dynamic during sporo-genesis with the formation of actin perinuclear shells each cell cycle and the emergence of polygonal territories during cellularization. Spizellomyces is a fast-growing and genetically-tractable organism that should be useful for comparative cell biology and understanding the evolution of fungi and early eukaryotes.

cell biology↗