Enabling neighbour labelling: using synthetic biology to explore how cells influence their neighbours

Development, Год журнала: 2024, Номер 151(1)

Опубликована: Янв. 1, 2024

ABSTRACT Cell-cell interactions are central to development, but exploring how a change in any given cell relates changes the neighbour of that can be technically challenging. Here, we review recent developments synthetic biology and image analysis helping overcome this problem. We highlight opportunities presented by these advances discuss limitations applying them developmental model systems.

Язык: Английский

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Constitutive and conditional epitope-tagging of endogenous G protein coupled receptors inDrosophila

Journal of Neuroscience, Год журнала: 2024, Номер 44(33), С. e2377232024 - e2377232024

Опубликована: Июнь 27, 2024

To visualize the cellular and subcellular localization of neuromodulatory G-protein–coupled receptors in Drosophila , we implement a molecular strategy recently used to add epitope tags ionotropic at their endogenous loci. Leveraging evolutionary conservation identify sites more likely permit insertion tag, generated constitutive conditional tagged alleles for 5-HT1A 5-HT2A 5-HT2B Oct β 1R 2R two isoforms OAMB mGluR . The allow restricted expression receptor specific cell types, an option not available any previous reagents label these proteins. We show patterns female brains that localize mushroom bodies (MBs) central complex, respectively, as predicted by roles sleep. By contrast, unexpected enrichment Octβ1R complex nerve terminals lobular columnar cells visual system suggest new hypotheses about functions sites. Using additional allele serotonin transporter, marker serotonergic tracts, demonstrate diverse spatial relationships between postsynaptic 5-HT presynaptic neurons, consistent with importance both synaptic volume transmission. Finally, use it localizes distinct within MBs Kenyon autoreceptor.

Язык: Английский

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Synaptic connectome of a neurosecretory network in the Drosophila brain

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

Опубликована: Авг. 29, 2024

Abstract Hormones mediate inter-organ signaling which is crucial in orchestrating diverse behaviors and physiological processes including sleep activity, feeding, growth, metabolism reproduction. The pars intercerebralis lateralis insects represent major hubs contain neurosecretory cells (NSC) that produce various hormones. To obtain insight into how hormonal regulated, we have characterized the synaptic connectome of NSC adult Drosophila brain. Identification neurons providing inputs to multiple subtypes implicates diuretic hormone 44-expressing as a coordinator physiology behavior. Surprisingly, despite most having dendrites subesophageal zone (primary taste processing center), gustatory are largely indirect. We also deciphered pathways via olfactory relayed NSC. Further, our analyses revealed substantial from descending NSC, suggesting regulate both endocrine motor output synchronize changes with appropriate behaviors. In contrast inputs, sparse mostly mediated by corazonin Therefore, additionally determine putative paracrine interconnectivity between peripheral tissues analyzing single-cell transcriptomic datasets. Our comprehensive characterization network provides platform understand complex networks they orchestrate animal physiology.

Язык: Английский

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PUFFFIN: an ultra-bright, customisable, single-plasmid system for labelling cell neighbourhoods

The EMBO Journal, Год журнала: 2024, Номер 43(18), С. 4110 - 4135

Опубликована: Июль 12, 2024

Cell communication coordinates developmental processes, maintains homeostasis, and contributes to disease. Therefore, understanding the relationship between cells in a shared environment is crucial. Here we introduce Positive Ultra-bright Fluorescent Fusion For Identifying Neighbours (PUFFFIN), cell neighbour-labelling system based upon secretion uptake of positively supercharged fluorescent protein s36GFP. We fused s36GFP mNeonGreen or HaloTag, facilitating ultra-bright, sensitive, colour-of-choice labelling. Secretor transfer PUFFFIN neighbours while retaining nuclear mCherry, making identification, isolation, investigation live straightforward. can be delivered cells, tissues, embryos on customisable single-plasmid construct composed interchangeable components with option incorporate any transgene. This versatility enables manipulation properties, simultaneously labelling surrounding culture vivo. use ask whether pluripotent adjust pace differentiation synchronise their during exit from naïve pluripotency. offers simple, approach profile non-cell-autonomous responses natural induced changes identity behaviour.

Язык: Английский

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Impact of developmental temperature on neural growth, connectivity, and function

Science Advances, Год журнала: 2025, Номер 11(3)

Опубликована: Янв. 15, 2025

Environmental temperature dictates the developmental pace of poikilothermic animals. In Drosophila , slower development at lower temperatures results in higher brain connectivity, but generality such scaling across and regions its impact on function are unclear. Here, we show that connectivity scales continuously temperatures, agreement with a first-principle model postulates different metabolic constraints for growth organism. The predicts wiring under cycles nonuniform temporal neural temperatures. Developmental has notable effects odor-driven behavior. Dissecting circuit architecture neurons olfactory pathway, demonstrate does not alter odor encoding first- second-order neurons, it shifts specificity connections onto third-order mediate innate behaviors. We conclude while some computations robust to wiring, others exhibit phenotypic plasticity possible adaptive advantages.

Язык: Английский

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Functional labeling of individualized postsynaptic neurons using optogenetics and trans-Tango in Drosophila (FLIPSOT)

PLoS Genetics, Год журнала: 2024, Номер 20(3), С. e1011190 - e1011190

Опубликована: Март 14, 2024

A population of neurons interconnected by synapses constitutes a neural circuit, which performs specific functions upon activation. It is essential to identify both anatomical and functional entities circuits comprehend the components processes necessary for healthy brain function changes that characterize disorders. To date, few methods are available study these two aspects circuit simultaneously. In this study, we developed FLIPSOT, or labeling individualized postsynaptic using optogenetics trans- Tango. FLIPSOT uses (1) Tango access genetically, (2) optogenetic approaches activate (FLIPSOTa) inhibit (FLIPSOTi) in random sparse manner, (3) fluorescence markers tagged with genes visualize neurons. Therefore, allows presynaptic driver behavioral individual readily applied has potential be adapted use mammalian circuits.

Язык: Английский

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Synaptic connectome of a neurosecretory network in the Drosophila brain

Опубликована: Ноя. 14, 2024

Hormones mediate inter-organ signaling which is crucial in orchestrating diverse behaviors and physiological processes including sleep activity, feeding, growth, metabolism reproduction. The pars intercerebralis lateralis insects represent major hubs contain neurosecretory cells (NSC) that produce various hormones. To obtain insight into how hormonal regulated, we have characterized the synaptic connectome of NSC adult Drosophila brain. Identification neurons providing inputs to multiple subtypes implicates diuretic hormone 44-expressing as a coordinator physiology behavior. Surprisingly, despite most having dendrites subesophageal zone (primary taste processing center), gustatory are largely indirect. We also deciphered pathways via olfactory relayed NSC. Further, our analyses revealed substantial from descending NSC, suggesting regulate both endocrine motor output synchronize changes with appropriate behaviors. In contrast inputs, sparse mostly mediated by corazonin Therefore, additionally determine putative paracrine interconnectivity between peripheral tissues analyzing single-cell transcriptomic datasets. Our comprehensive characterization network provides platform understand complex networks they orchestrate animal physiology.

Язык: Английский

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Molecular characterization of gustatory second-order neurons reveals integrative mechanisms of gustatory and metabolic information

Опубликована: Ноя. 8, 2024

Animals must balance the urgent need to find food during starvation with critical necessity avoid toxic substances ensure their survival. In Drosophila , specialized Gustatory Receptors (GRs) expressed in Receptor Neurons (GRNs) are for distinguishing between nutritious and potentially food. GRNs project axons from taste organs Subesophageal Zone (SEZ) Central Brain (CB) of where gustatory information is processed. Although roles GRs well- documented, processing SEZ remains unclear. To better understand sensory feeding decision-making, we molecularly characterized first layer interneurons, referred as Second Order (G2Ns), which receive direct input GRNs. Using trans-synaptic tracing trans- Tango, cell sorting, bulk RNAseq under fed starved conditions, discovered that G2Ns vary based on molecular profile changes fly’s metabolic state. Further data analysis has revealed a pair neurons SEZ, expressing neuropeptide Leucokinin (SELK neurons), simultaneous sensing bitter (potentially toxic) sweet (nutritious) information. Additionally, these also inputs regarding levels fly. These results highlight novel mechanism regulation integration.

Язык: Английский

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Drosulfakinin signaling encodes early-life memory for adaptive social plasticity

eLife, Год журнала: 2024, Номер 13

Опубликована: Дек. 18, 2024

Drosophila establishes social clusters in groups, yet the underlying principles remain poorly understood. Here, we performed a systemic analysis of network behavior (SNB) that quantifies individual distance (SD) group over time. The SNB assessment 175 inbred strains from Genetics Reference Panel showed tight association short SD with long developmental time, low food intake, and hypoactivity. inferiority short-SD individuals was compensated by their culturing. By contrast, isolation silenced beneficial effects interactions adults blunted plasticity under physiological challenges. Transcriptome analyses revealed genetic diversity for traits, whereas reprogrammed select pathways, regardless phenotypes. In particular, deprivation suppressed expression neuropeptide Drosulfakinin ( Dsk ) three pairs adult brain neurons. Male-specific DSK signaling to cholecystokinin-like receptor 17D1 mediated plasticity. fact, transgenic manipulations neuron activity were sufficient imitate state experience. Given functional conservation mammalian homologs, propose animals may have evolved dedicated neural mechanism encode early-life experience transform properties adaptively.

Язык: Английский

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Transsynaptic labeling and transcriptional control of zebrafish neural circuits

Nature Neuroscience, Год журнала: 2024, Номер 28(1), С. 189 - 200

Опубликована: Дек. 19, 2024

Язык: Английский

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