scholarly journals The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging

10.3791/56101 ◽  
2017 ◽  
Author(s):  
Nan Zhang ◽  
Edward Membreno ◽  
Susan Raj ◽  
Hongjie Zhang ◽  
Liakot A Khan ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Labeling ◽  
Membrane Biogenesis ◽  
C Elegans ◽  
Excretory Canal

scholarly journals Combining single-cell RNA-sequencing with a molecular atlas unveils new markers for Caenorhabditis elegans neuron classes

2020 ◽  
Author(s):  
Ramiro Lorenzo ◽  
Michiho Onizuka ◽  
Matthieu Defrance ◽  
Patrick Laurent
Keyword(s):  
Caenorhabditis Elegans ◽  
Single Cell ◽  
Rna Sequencing ◽  
Cell Fate ◽  
Single Neuron ◽  
Genetic Manipulations ◽  
C Elegans ◽  
Single Cell Rna Sequencing ◽  
Normal Differentiation

Abstract Single-cell RNA-sequencing (scRNA-seq) of the Caenorhabditis elegans nervous system offers the unique opportunity to obtain a partial expression profile for each neuron within a known connectome. Building on recent scRNA-seq data and on a molecular atlas describing the expression pattern of ∼800 genes at the single cell resolution, we designed an iterative clustering analysis aiming to match each cell-cluster to the ∼100 anatomically defined neuron classes of C. elegans. This heuristic approach successfully assigned 97 of the 118 neuron classes to a cluster. Sixty two clusters were assigned to a single neuron class and 15 clusters grouped neuron classes sharing close molecular signatures. Pseudotime analysis revealed a maturation process occurring in some neurons (e.g. PDA) during the L2 stage. Based on the molecular profiles of all identified neurons, we predicted cell fate regulators and experimentally validated unc-86 for the normal differentiation of RMG neurons. Furthermore, we observed that different classes of genes functionally diversify sensory neurons, interneurons and motorneurons. Finally, we designed 15 new neuron class-specific promoters validated in vivo. Amongst them, 10 represent the only specific promoter reported to this day, expanding the list of neurons amenable to genetic manipulations.

scholarly journals Combining single-cell RNA-sequencing with a molecular atlas unveils new markers for C. elegans neuron classes

2019 ◽  
Author(s):  
Ramiro Lorenzo ◽  
Michiho Onizuka ◽  
Matthieu Defrance ◽  
Patrick Laurent
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Motor Neurons ◽  
Clustering Analysis ◽  
Molecular Signatures ◽  
Functional Diversification ◽  
Genetic Manipulations ◽  
C Elegans ◽  
Single Cell Rna Sequencing

AbstractSingle-cell RNA-sequencing (scRNA-seq) of the Caenorhabditis elegans (C. elegans) nervous system offers the unique opportunity to obtain a partial expression profile for each neuron within a known connectome. Building on recent scRNA-seq data [1] and on a molecular atlas describing the expression pattern of ~800 genes at the single cell resolution [2], we designed an iterative clustering analysis aiming to match each cell-cluster to the ~100 anatomically defined neuron classes of C. elegans. This heuristic approach successfully assigned 58 clusters to their corresponding neuron class. Another 11 clusters grouped neuron classes sharing close molecular signatures and 7 clusters were not assigned. Based on these 76 molecular profiles, we designed 15 new neuron class-specific promoters validated in vivo. Amongst them, 10 represent the only specific promoter reported to this day, expanding the list of neurons amenable to genetic manipulations. Finally, we observed a differential expression of functionally relevant genes between sensory-, inter-, and motor neurons in C. elegans, suggesting the mode of functional diversification may vary accordingly to the neuronal modalities.

scholarly journals Probing Single-Cell Micromechanics In Vivo: The Microrheology of C. elegans Developing Embryos

2006 ◽  
Vol 90 (12) ◽  
pp. 4712-4719 ◽  
Author(s):  
Brian R. Daniels ◽  
Byron C. Masi ◽  
Denis Wirtz
Keyword(s):  
Single Cell ◽  
C Elegans ◽  
Cell Micromechanics

scholarly journals Integration of plasticity mechanisms within a single sensory neuron of C. elegans actuates a memory

2017 ◽  
Author(s):  
Josh D. Hawk ◽  
Ana C. Calvo ◽  
Agustin Almoril-Porras ◽  
Ahmad Aljobeh ◽  
Maria Luisa Torruella-Suárez ◽  
...  
Keyword(s):  
Single Cell ◽  
Sensory Neuron ◽  
Neural Plasticity ◽  
Sensory Information ◽  
Sensory Adaptation ◽  
Sensory Stimuli ◽  
C Elegans ◽  
Presynaptic Plasticity ◽  
Behavioral Preference

SummaryNeural plasticity—the ability of a neuron to change its cellular properties in response to past experiences—underpins the nervous system’s capacity to form memories and actuate behaviors. How different plasticity mechanisms act together in vivo and at a cellular level to transform sensory information into behavior is not well understood. Here we show that in the nematode C. elegans two plasticity mechanisms—sensory adaptation and presynaptic plasticity—act within a single cell to encode thermosensory information and actuate a temperature-preference memory. Sensory adaptation enables the primary thermosensory neuron, AFD, to adjust the temperature range of its sensitivity to the local environment, thereby optimizing its ability to detect temperature fluctuations associated with migration. Presynaptic plasticity transforms this thermosensory information into a behavioral preference by gating synaptic communication between sensory neuron AFD and its postsynaptic partner, AIY. The gating of synaptic communication is regulated at AFD presynaptic sites by the conserved kinase nPKCε. Bypassing or altering AFD presynaptic plasticity predictably changes the learned behavioral preferences without affecting sensory responses. Our findings indicate that two distinct and modular neuroplasticity mechanisms function together within a single sensory neuron to encode multiple components of information required to enact thermotactic behavior. The integration of these plasticity mechanisms result in a single-cell logic system that can both represent sensory stimuli and guide memory-based behavioral preference.

scholarly journals Supernova: A Versatile Vector System for Single-Cell Labeling and Gene Function Studies in vivo

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Wenshu Luo ◽  
Hidenobu Mizuno ◽  
Ryohei Iwata ◽  
Shingo Nakazawa ◽  
Kosuke Yasuda ◽  
...  
Keyword(s):  
Single Cell ◽  
Gene Function ◽  
Cell Labeling ◽  
Vector System

scholarly journals In vivo single-cell lineage tracing in zebrafish using high-resolution infrared laser-mediated gene induction microscopy

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Sicong He ◽  
Ye Tian ◽  
Shachuan Feng ◽  
Yi Wu ◽  
Xinwei Shen ◽  
...  
Keyword(s):  
Single Cell ◽  
Zebrafish Embryo ◽  
Cell Lineage ◽  
Cell Types ◽  
Infrared Laser ◽  
Lineage Tracing ◽  
Cell Labeling ◽  
Specific Cell ◽  
Blood Island

Heterogeneity broadly exists in various cell types both during development and at homeostasis. Investigating heterogeneity is crucial for comprehensively understanding the complexity of ontogeny, dynamics, and function of specific cell types. Traditional bulk-labeling techniques are incompetent to dissect heterogeneity within cell population, while the new single-cell lineage tracing methodologies invented in the last decade can hardly achieve high-fidelity single-cell labeling and long-term in-vivo observation simultaneously. In this work, we developed a high-precision infrared laser-evoked gene operator heat-shock system, which uses laser-induced CreERT2 combined with loxP-DsRedx-loxP-GFP reporter to achieve precise single-cell labeling and tracing. In vivo study indicated that this system can precisely label single cell in brain, muscle and hematopoietic system in zebrafish embryo. Using this system, we traced the hematopoietic potential of hemogenic endothelium (HE) in the posterior blood island (PBI) of zebrafish embryo and found that HEs in the PBI are heterogeneous, which contains at least myeloid unipotent and myeloid-lymphoid bipotent subtypes.

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