Vectors for co-expression of two genes in Caenorhabditis elegans

Lee, Li‐Wei; Lo, Hsiao-Wen; Lo, Szecheng J.

doi:10.1016/j.gene.2010.02.001

Cited by 13 publications

(12 citation statements)

References 17 publications

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“…** P < 0.02, as determined by a Wilcoxon signed-rank test. (D) Measurement of number of nucleoli per cell in ASEL and ASER using a translational FIB-1::GFP reporter [52]. ASE neurons were identified with a che-1 prom :: mChOpti transgene that labels ASE neurons in red.…”

Section: Resultsmentioning

confidence: 99%

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Developmental control of lateralized neuron size in the nematode Caenorhabditis elegans

et al. 2010

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BackgroundNervous systems are generally bilaterally symmetric on a gross structural and organizational level but are strongly lateralized (left/right asymmetric) on a functional level. It has been previously noted that in vertebrate nervous systems, symmetrically positioned, bilateral groups of neurons in functionally lateralized brain regions differ in the size of their soma. The genetic mechanisms that control these left/right asymmetric soma size differences are unknown. The nematode Caenorhabditis elegans offers the opportunity to study this question with single neuron resolution. A pair of chemosensory neurons (ASEL and ASER), which are bilaterally symmetric on several levels (projections, synaptic connectivity, gene expression patterns), are functionally lateralized in that they express distinct chemoreceptors and sense distinct chemosensory cues.ResultsWe describe here that ASEL and ASER also differ substantially in size (soma volume, axonal and dendritic diameter), a feature that is predicted to change the voltage conduction properties of the two sensory neurons. This difference in size is not dependent on sensory input or neuronal activity but developmentally programmed by a pathway of gene regulatory factors that also control left/right asymmetric chemoreceptor expression of the two ASE neurons. This regulatory pathway funnels via the DIE-1 Zn finger transcription factor into the left/right asymmetric distribution of nucleoli that contain the rRNA regulator Fibrillarin/FIB-1, a RNA methyltransferase implicated in the non-hereditary immune disease scleroderma, which we find to be essential to establish the size differences between ASEL and ASER.ConclusionsTaken together, our findings reveal a remarkable conservation of the linkage of functional lateralization with size differences across phylogeny and provide the first insights into the developmentally programmed regulatory mechanisms that control neuron size lateralities.

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Section: Resultsmentioning

confidence: 99%

“…To determine nucleoli size and number, we used cguIs001 ( fib-1::gfp ) [52] and an antibody against Nop1p (FIB-1) from EnCor BioTechnology (#MCA-38F3, Gainesville, FL, USA) at a 1:200 dilution, detected with a 1:200 dilution of an anti-mouse (Invitrogen #A-21202, Carlsbad, CA, USA") secondary antibody.…”

Section: Methodsmentioning

confidence: 99%

Developmental control of lateralized neuron size in the nematode Caenorhabditis elegans

et al. 2010

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“…Lo, was modified by first removing the myo-2 promoter and L-HDAg gene within, then replacing the original unc-54 3′UTR with unc-1 3′UTR. The Dual Reporter (DR) module in pDR consists of gfp::icr::rfp as reported (27). DR reporters were subsequently generated from pDR as follows:…”

Section: Methodsmentioning

confidence: 99%

A Steroid Receptor–MicroRNA Switch Regulates Life Span in Response to Signals from the Gonad

Shen

Wollam

Magner

et al. 2012

Science

103

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While the gonad primarily functions in procreation, it also affects animal lifespan. Here we show that removal of the C. elegans germline triggers a switch in the regulatory state of the organism to promote longevity, co-opting components involved in larval developmental timing circuits. These include the DAF-12 steroid receptor, which upregulates members of the let-7 microRNA family, involved in L2/L3 transitions. The microRNAs target an early larval nuclear factor, lin-14, and akt-1/kinase, thereby stimulating DAF-16/FOXO signaling to extend life. These studies suggest that metazoan lifespan is coupled to the gonad through elements of a developmental timer.

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“…LDAg is detectable at the nucleoli of transfected human hepatoma cells by using GFP as a reporter of gene expression (Figures 6(a)–6(c)). When transgenic worms coexpress FIB-1::GFP and RFP::LD using a bicistronic vector [59], LDAg is localized at the nucleoli of most cells by FIB-1::GFP (Figures 6(d)–6(f)). Interestingly, in the presence of FIB-1::GFP, RFP::LD is found at the perimeter of the nucleolus in intestinal cells (Figures 6(g)–6(i)).…”

Section: Morphology Of C Elegans Nucleolimentioning

confidence: 99%

The Nucleolus ofCaenorhabditis elegans

Lee

Lee²,

Huang³

et al. 2012

Journal of Biomedicine and Biotechnology

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Nucleolar size and appearance correlate with ribosome biogenesis and cellular activity. The mechanisms underlying changes in nucleolar appearance and regulation of nucleolar size that occur during differentiation and cell cycle progression are not well understood. Caenorhabditis elegans provides a good model for studying these processes because of its small size and transparent body, well-characterized cell types and lineages, and because its cells display various sizes of nucleoli. This paper details the advantages of using C. elegans to investigate features of the nucleolus during the organism's development by following dynamic changes in fibrillarin (FIB-1) in the cells of early embryos and aged worms. This paper also illustrates the involvement of the ncl-1 gene and other possible candidate genes in nucleolar-size control. Lastly, we summarize the ribosomal proteins involved in life span and innate immunity, and those homologous genes that correspond to human disorders of ribosomopathy.

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Vectors for co-expression of two genes in Caenorhabditis elegans

Cited by 13 publications

References 17 publications

Developmental control of lateralized neuron size in the nematode Caenorhabditis elegans

Developmental control of lateralized neuron size in the nematode Caenorhabditis elegans

A Steroid Receptor–MicroRNA Switch Regulates Life Span in Response to Signals from the Gonad

The Nucleolus ofCaenorhabditis elegans

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