Cell isolation and culture

Zhang, Sihui; Kuhn, Jeffrey R.

doi:10.1895/wormbook.1.157.1

Cited by 35 publications

(33 citation statements)

References 85 publications

(28 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To characterize the molecular mechanisms through which amphid sheath (AMsh) glia regulate neuronal function, and not development, we took advantage of recently developed larval cell isolation techniques and adapted these to identify AMsh glia post-embryonic transcripts by RNA sequencing (Spencer et al, 2014; Zhang and Kuhn, 2013). This approach led to the identification of 598 candidate AMsh glial genes.…”

Section: Discussionmentioning

confidence: 99%

“…After growth for 36-42 hours, cells were isolated from late-stage larvae (L3 and L4) by protease treatment and mechanical disruption, as described previously (Zhang and Kuhn, 2013), with modifications (Menachem Katz, personal communications). AMsh glia expressing a dsRed transgene ( nsIs143 ) were sorted using a BD FACS Aria sorter (Rockefeller University Flow Cytometry Resource Center).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

PROS-1/Prospero Is a Major Regulator of the Glia-Specific Secretome Controlling Sensory-Neuron Shape and Function in C. elegans

et al. 2016

View full text Add to dashboard Cite

SUMMARY Sensory neurons are an animal’s gateway to the world, and their receptive endings, the sites of sensory signal transduction, are often associated with glia. While glia are known to promote sensory-neuron functions, the molecular bases of these interactions are poorly explored. Here we describe a post-developmental glial role for the PROS-1/Prospero/PROX1 homeodomain protein in sensory-neuron function in C. elegans. Using glia expression profiling, we demonstrate that, unlike previously characterized cell fate roles, PROS-1 functions post-embryonically to control sense-organ glia-specific secretome expression. PROS-1 functions cell autonomously to regulate glial secretion and membrane structure, and non-cell autonomously to control the shape and function of the receptive endings of sensory neurons. Known glial genes controlling sensory-neuron function are PROS-1 targets, and we identify additional PROS-1-dependent genes required for neuron attributes. Drosophila Prospero and vertebrate PROX1 are expressed in post-mitotic sense-organ glia and in astrocytes, suggesting conserved roles for this class of transcription factors.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

PROS-1/Prospero Is a Major Regulator of the Glia-Specific Secretome Controlling Sensory-Neuron Shape and Function in C. elegans

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Given that the protease zmp-1 is expressed throughout AC invasion, zmp-1 and other proteases might assist invasion by weakening the structural make-up of the basement membrane to facilitate physical displacement. Further sensitized screening, as well as profiling gene expression in the AC using single cell isolation techniques, 116 will help reveal mechanisms of how basement membrane is modified to assist in cell invasion events.…”

Section: Invadopodia Are Only One Component Of the Invasion Process Imentioning

confidence: 99%

Invadopodia and basement membrane invasion in vivo

Lohmer

Kelley

Hagedorn

et al. 2014

Cell Adhesion & Migration

View full text Add to dashboard Cite

“…Tissue-specific gene expression profiling has been very successful in C. elegans embryos because it is relatively easy to dissociate embryos for cell sorting (5). However, larvae and adults have a tough cuticle barrier preventing cell isolation.…”

Section: Introductionmentioning

confidence: 99%

Analysis ofC. elegansmuscle transcriptome using trans-splicing-based RNA tagging (SRT)

Zhan

Sleumer

et al. 2016

Nucleic Acids Res

View full text Add to dashboard Cite

Current approaches to profiling tissue-specific gene expression in C. elegans require delicate manipulation and are difficult under certain conditions, e.g. from dauer or aging worms. We have developed an easy and robust method for tissue-specific RNA-seq by taking advantage of the endogenous trans-splicing process. In this method, transgenic worms are generated in which a spliced leader (SL) RNA gene is fused with a sequence tag and driven by a tissue-specific promoter. Only in the tissue of interest, the tagged SL RNA gene is transcribed and then trans-spliced onto mRNAs. The tag allows enrichment and sequencing of mRNAs from that tissue only. As a proof of principle, we profiled the muscle transcriptome, which showed high coverage and efficient enrichment of muscle specific genes, with low background noise. To demonstrate the robustness of our method, we profiled muscle gene expression in dauer larvae and aging worms, revealing gene expression changes consistent with the physiology of these stages. The resulting muscle transcriptome also revealed 461 novel RNA transcripts, likely muscle-expressed long non-coding RNAs. In summary, the splicing-based RNA tagging (SRT) method provides a convenient and robust tool to profile trans-spliced genes and identify novel transcripts in a tissue-specific manner, with a low false positive rate.

show abstract

Cell isolation and culture

Cited by 35 publications

References 85 publications

PROS-1/Prospero Is a Major Regulator of the Glia-Specific Secretome Controlling Sensory-Neuron Shape and Function in C. elegans

PROS-1/Prospero Is a Major Regulator of the Glia-Specific Secretome Controlling Sensory-Neuron Shape and Function in C. elegans

Invadopodia and basement membrane invasion in vivo

Analysis ofC. elegansmuscle transcriptome using trans-splicing-based RNA tagging (SRT)

Contact Info

Product

Resources

About