Cell-specific microarray profiling experiments reveal a comprehensive picture of gene expression in the C. elegans nervous system

Stetina, Stephen E. Von; Watson, Joseph D.; Fox, Rebecca; Olszewski, Kellen L.; Spencer, W. Clay; Roy, Peter J.; Miller, David M.

doi:10.1186/gb-2007-8-7-r135

Cited by 112 publications

(144 citation statements)

References 109 publications

(128 reference statements)

Supporting

Mentioning

135

Contrasting

Order By: Relevance

“…We also report published representative results obtained using a variety of techniques including immunocytochemistry, electrophysiology and calcium imaging that have advanced our understanding of the physiological processes and genetic requirements underlying cell function in C. elegans. Work done so far on cultured C. elegans cells supports that they differentiate in vitro as they do in vivo recapitulating the expression of cell specific markers 1,[18][19][20]22,23,27,44 . While not all cell types have been characterized in culture, this body of work suggests that the cell culture method allows the study of C. elegans cells in isolation without compromising their identity.…”

Section: Discussionmentioning

confidence: 99%

“…elegans cells in vitro 1 , as well as isolation of specific cell types by Fluorescent-Activated Cell Sorting (FACS) for the construction of cell-specific cDNA libraries 22,23 . Gene knockdown techniques such as RNA interference (RNAi) can be applied on cultured C. elegans cells 1 and a novel metabolic labeling method using Azido-sugar as a tool for glycoprotein discovery has been recently developed for in vitro cultured C. elegans cells 24 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A Method for Culturing Embryonic <em>C. elegans</em> Cells

Sangaletti

Bianchi

2013

JoVE

View full text Add to dashboard Cite

C. elegans is a powerful model system, in which genetic and molecular techniques are easily applicable. Until recently though, techniques that require direct access to cells and isolation of specific cell types, could not be applied in C. elegans. This limitation was due to the fact that tissues are confined within a pressurized cuticle which is not easily digested by treatment with enzymes and/or detergents. Based on early pioneer work by Laird Bloom, Christensen and colleagues 1 developed a robust method for culturing C. elegans embryonic cells in large scale. Eggs are isolated from gravid adults by treatment with bleach/NaOH and subsequently treated with chitinase to remove the eggshells. Embryonic cells are then dissociated by manual pipetting and plated onto substrate-covered glass in serum-enriched media. Within 24 hr of isolation cells begin to differentiate by changing morphology and by expressing cell specific markers. C. elegans cells cultured using this method survive for up 2 weeks in vitro and have been used for electrophysiological, immunochemical, and imaging analyses as well as they have been sorted and used for microarray profiling. Video LinkThe video component of this article can be found at

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Method for Culturing Embryonic <em>C. elegans</em> Cells

Sangaletti

Bianchi

2013

JoVE

View full text Add to dashboard Cite

show abstract

“…They achieved this by expressing EGFP under the promoter control of a gene, which was uniquely expressed in the desired cell population. [44][45][46][47][48] Another significant drawback to defining cells of interest by their endogenous cell surface markers is that these markers may not be expressed at the earliest stages of differentiation. Onset of stem cell differentiation down a particular lineage is often defined by a transcription factor functioning as a master regulator.…”

Section: Discussionmentioning

confidence: 99%

Development of the “Three-step MACS”: a Novel Strategy for Isolating Rare Cell Populations in the Absence of Known Cell Surface Markers from Complex Animal Tissue

Lee¹,

Lufkin²

2012

J Biomol Tech

View full text Add to dashboard Cite

To circumvent the difficulty of isolating specific cell populations by MACS from dissociated complex animal tissue, when their proportions reached levels similar to that of the background, we developed the "Three-step MACS" strategy. Cells of interest are defined by their expression of a particular gene(s) of interest rather by than their natural cell surface markers or size. A two-component transgenic cell surface protein, for two sequential rounds of MACS, is expressed under the promoter control of the endogenous gene of interest by means of gene targeting and the generation of transgenic tissue. An initial step to remove dead cells is also used. Here, we describe proof-of-concept experiments, using the biotin acceptor peptide (BAP)-low-affinity nerve growth factor receptor as the two-component protein. The first component, the BAP, can be biotinylated in specific subsets of cells expressing a particular gene by expressing the biotinylating enzyme, hBirA ϭ humanized BirA (hBirA), under the promoter control of another gene defining the specific subpopulation. We showed that a rare population of cells (1.1% of the 13.5 days postcoital mouse embryo) could be enriched to a sufficiently high purity (84.4%). From another sample with 0.1% of our cells of interest, we achieved a 40.3% pure sample. The low cost, speed, and technical ease of the Three-step MACS also make it scalable and hence, an ideal method for preparing sufficient quantities of biological samples for sensitive, high-throughput assays.

show abstract

“…The silencing of this expression potentially helps the scientific community better understand the function of the gene's product [16][17][18]. Extensive experimentation with C. elegans has produced complete neuronal and developmental wiring diagrams that have been confirmed by numerous labs [3,19]. Furthermore, these networks are known to be similar in humans, providing additional justification for comparing two widely diverse organisms [5].…”

mentioning

confidence: 99%