Perisomatic ultrastructure efficiently classifies cells in mouse cortex

Nature, Год журнала: 2025, Номер 640(8058), С. 478 - 486

Опубликована: Апрель 9, 2025

Abstract Mammalian neocortex contains a highly diverse set of cell types. These types have been mapped systematically using variety molecular, electrophysiological and morphological approaches 1–4 . Each modality offers new perspectives on the variation biological processes underlying cell-type specialization. Cellular-scale electron microscopy provides dense ultrastructural examination an unbiased perspective subcellular organization brain cells, including their synaptic connectivity nanometre-scale morphology. In data that contain tens thousands neurons, most which incomplete reconstructions, identifying becomes clear challenge for analysis 5 Here, to address this challenge, we present systematic survey somatic region all cells in cubic millimetre cortex quantitative features obtained from microscopy. This demonstrates perisomatic is sufficient identify types, defined primarily basis patterns. We then describe how classification facilitates cell-type-specific characterization locating with rare patterns dataset.

Язык: Английский

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Permanent deconstruction of intracellular primary cilia in differentiating granule cell neurons

The Journal of Cell Biology, Год журнала: 2024, Номер 223(10)

Опубликована: Июнь 27, 2024

Primary cilia on granule cell neuron progenitors in the developing cerebellum detect sonic hedgehog to facilitate proliferation. Following differentiation, cerebellar cells become most abundant neuronal type brain. While are essential during early developmental stages, they infrequent upon maturation. Here, we provide nanoscopic resolution of situ using large-scale electron microscopy volumes and immunostaining mouse cerebella. In many cells, found intracellular cilia, concealed from external environment. Cilia were disassembled differentiating neurons-in a process call deconstruction-distinct premitotic resorption proliferating progenitors. deconstruction involved unique disassembly intermediates, and, as maturation progressed, mother centriolar docking at plasma membrane. Unlike ciliated neurons other brain regions, our results show which might prevent mitogenic responsiveness. Ciliary could be paradigmatic removal differentiation tissues.

Язык: Английский

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Structure, interaction and nervous connectivity of beta cell primary cilia

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Окт. 24, 2024

Primary cilia are sensory organelles present in many cell types, partaking various signaling processes. of pancreatic beta cells play pivotal roles paracrine and their dysfunction is linked to diabetes. Yet, the structural basis for functions unclear. We three-dimensional reconstructions primary by electron expansion microscopy. These spatially confined within deep ciliary pockets or narrow spaces between cells, lack motility components display an unstructured axoneme organization. Furthermore, we observe a plethora cilia-cilia cilia-cell interactions with other islet non-islet cells. Most remarkably, have identified characterized axo-ciliary synapses cholinergic innervation. findings highlight cilia's role connectivity, pointing at function integrating intrinsic extrinsic signals contribute understanding significance health

Язык: Английский

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Differences in neuronal ciliation rate and ciliary content revealed by systematic imaging-based analysis of hiPSC-derived models across protocols

Frontiers in Cell and Developmental Biology, Год журнала: 2025, Номер 13

Опубликована: Апрель 11, 2025

Introduction Ciliopathies are a group of human Mendelian disorders caused by dysfunction primary cilia, small quasi-ubiquitous sensory organelles. Patients suffering from ciliopathies often display prominent neurodevelopmental phenotypes, underscoring the importance cilia during development and for function central nervous system (CNS). Human tissues, in particular CNS, very hard to obtain research. Patient derived- or genetically engineered induced pluripotent stem cells (hiPSCs) therefore precious resource investigating role neurons. Methods In this study we used variety 2D 3D neuronal differentiation protocols multiple hiPSC lines systematically analyzed ciliation rates ciliary length hiPSCs, neural (NSCs), immature different types mature neurons using immunofluorescence. Results We found that rate varied substantially between cell protocols. Moreover, depended on stage, being maximal NSCs decreasing with maturation. various obtained protocols, be as low ∼10%. Neuronal density also played an important role, higher denser cultures. further investigated protein content these at stages commonly antibodies against ARL13B, INPP5E, AC3 GPR161. Cilia were all positive trend intensity more Likewise, INPP5E was present analyzed, while positivity increased maturation proceeded. Interestingly, GPR161 signal almost completely disappeared upon Sonic hedgehog (SHH) stimulation neurons, not case suggesting possible developmental time window cilia-dependent SHH signaling. Conclusion Taken together, our results provide systematic description hiPSC-derived generated selecting optimal model controls cells.

Язык: Английский

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Methods to Assess Neuronal Primary Cilia Electrochemical Signaling

Journal of Cellular Physiology, Год журнала: 2025, Номер 240(4)

Опубликована: Апрель 1, 2025

ABSTRACT Primary cilia are polymodal sensory organelles which project from the apical side of polarized cells. They found in all brain hemispheres but most pronounced neurons, comprise granular layers hippocampus and cerebellum. Pathogenic variants genes encode primary components responsible for neuronal ciliopathies—a group central nervous system disorders characterized by neurodevelopmental conditions such as intellectual disability, seizure, ataxia, deficits. In hippocampus, form chemical synapses with axons their membranes populated unique sets ion channels G protein‐coupled receptors (GPCRs). small privileged compartments that challenging to study. detail, we describe electrophysiology methods use cilia‐specific fluorescent sensors assay polycystin channel function serotonergic receptor signaling, respectively. These tools allow researchers calcium, cAMP channel‐related signaling pathways isolated neurons real‐time semi‐quantitative terms, while enhancing our understanding this understudied organelle its dysregulation ciliopathy disease states.

Язык: Английский

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Permanent cilia loss during cerebellar granule cell neurogenesis involves withdrawal of cilia maintenance and centriole capping

Proceedings of the National Academy of Sciences, Год журнала: 2024, Номер 121(52)

Опубликована: Дек. 20, 2024

Brain neurons utilize the primary cilium as a privileged compartment to detect and respond extracellular ligands such Sonic hedgehog (SHH). However, cilia in cerebellar granule cell (GC) disassemble during differentiation through ultrastructurally unique intermediates, process we refer deconstruction. In addition, mature do not reciliate despite having docked centrioles. Here, identify molecular changes that accompany deconstruction centriole docking GC neurons. We used single transcriptomic immunocytological analyses compare transcript levels subcellular localization of proteins between progenitor, differentiating, GCs. Differentiating GCs lacked transcripts for key activators premitotic resorption, indicating disassembly differentiating cells is distinct from resorption. Instead, differentiation, many genes required maintenance—specifically those encoding components intraflagellar transport, pericentrosomal material, centriolar satellites—decreased. The abundance several corresponding around centrosomes also decreased. These coincided with downregulation SHH signaling prior even mutant excessive activation. Finally, mother centrioles maturing recruited cap complex protein, CEP97. data suggest global, developmentally programmed decrease maintenance mediates deconstruction, while capping prevents regrowth dysregulated signaling. Our study provides mechanistic insights expanding our understanding permanent loss multiple tissue-specific contexts.

Язык: Английский

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Single-cell transcriptomic changes in oligodendrocytes and precursors derived from Parkinson's disease patient-iPSCs with LRRK2-G2019S mutation

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июль 3, 2024

Abstract Despite extensive research, the contribution of LRRK2 p.G2019S mutation to Parkinson’s disease (PD) remains unclear. Recent findings indicate oligodendrocytes (ODCs) and their progenitors are vulnerable in PD pathogenesis. Notably, oligodendrocyte precursor cells (OPCs) exhibit high endogenous expression . We induced patient-iPSCs with into oligodendroglial lineages performed single-cell RNA sequencing. Cell type composition analysis revealed an increase OPCs, proliferating OPCs ciliated ependymal lines, all which characterized by expression. Differential transcriptomic changes several pathways, including down-regulation genes related myelin assembly ODCs, semaphorin-plexin pathway cilium movement OPCs. Cell-cell communication identified significant alterations signaling pathways a deactivation PSAP activation MIF lines. Additionally, we observed overall SEMA6 cell lines; however, derived from these lines specifically lost due SEMA6A PLXNA2 Pseudotemporal trajectory that SHH had significantly altered along pseudotime, accompanied higher levels propose dysfunctional signaling, cilia might represent early events pathology.

Язык: Английский

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Roles for primary cilia in synapses and neurological disorders

Trends in Cell Biology, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 1, 2024

Язык: Английский

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