The postembryonic cell lineages of the hermaphrodite and male gonads in Caenorhabditis elegans

Kimble, Judith; Hirsh, David

doi:10.1016/0012-1606(79)90035-6

Cited by 802 publications

(654 citation statements)

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“…Although the entire somatic cell lineage has been mapped in real time (Sulston and Horvitz 1977;Kimble and Hirsh 1979;Sulston et al 1983), following a cell lineage is often an arduous task that is made more difficult when mutant conditions perturb landmarks or alter cell identities. Our application of the FLP-out technology to lineaging offers an alternative whereby a cell lineage can be followed by virtue of an independent and permanent lineage mark.…”

Section: Discussionmentioning

confidence: 99%

A “FLP-Out” System for Controlled Gene Expression in Caenorhabditis elegans

Voutev

Hubbard

2008

Genetics

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We present a two-part system for conditional FLP-out of FRT-flanked sequences in Caenorhabditis elegans to control gene activity in a spatially and/or temporally regulated manner. Using reporters, we assess the system for efficacy and demonstrate its use as a cell lineage marking tool. In addition, we construct and test a dominant-negative form of hlh-12, a gene that encodes a basic helix-loop-helix (bHLH) transcription factor required for proper distal tip cell (DTC) migration. We show that this allele can be conditionally expressed from a heat-inducible FLP recombinase and can interfere with DTC migration. Using the same DTC assay, we conditionally express an hlh-12 RNAi-hairpin and induce the DTC migration defect. Finally, we introduce a set of traditional and Gateway-compatible vectors to facilitate construction of plasmids for this technology using any promoter, reporter, and gene/hairpin of interest.T WO-COMPONENT gene expression systems are indispensable tools to probe molecular mechanisms underlying development. Because control can be exerted by each component independently, exquisite temporal and spatial control of gene activation or repression can be achieved. Using different promoter combinations to drive each component of these systems, additional control can be obtained beyond that afforded by heat-inducible or tissue-specific promoters alone. Site-specific recombination systems such as the FLP/FRT system have been used to control gene expression by ''FLP-out'': a recombinase-catalyzed intramolecular excision of spacer DNA that lies between tandemly oriented FRT sites. The spacer includes a transcriptional stop so that prior to activation of the FLP recombinase (and subsequent FLP-out) the gene downstream of the spacer is not transcribed (Golic and Lindquist 1989;Struhl and Basler 1993; Figure 1A). After the FRT-containing cassette is excised by the FLP recombinase, the downstream gene is brought into proximity to the promoter and is expressed (reporter 2 in Figure 1A). This system and related systems have proven quite powerful and flexible in model organisms including Drosophila and mouse (see Branda and Dymecki 2004, for review;McGuire et al. 2004). However, prior to our study presented here, and a recently published study (Davis et al. 2008), these systems had not been developed for use in Caenorhabditis elegans.An ideal FLP-out system provides the means to generate both loss-and gain-of-function effects in a spatially and temporally controlled manner. In addition, wild-type gene expression can be turned on in particular cells at particular times in an otherwise mutant background. In organisms where transgenes can be reliably inserted in single copy, FLP-out can also be used to eliminate wild-type gene expression by excision of an FRT-flanked wild-type cassette in a mutant background. In C. elegans, the most common methods for generating transgenic C. elegans introduce multiple copies of transgenes on extrachromosomal arrays (Stinchcomb et al. 1985;Mello et al. 1991;Kelly et al. 1997). ...

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Section: Discussionmentioning

confidence: 99%

A “FLP-Out” System for Controlled Gene Expression in Caenorhabditis elegans

Voutev

Hubbard

2008

Genetics

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“…In the L2 stage in the C. elegans hermaphrodite, one of the somatic gonadal cells (Z1.ppp or Z4.aaa) moves into the central position on the ventral surface of the gonad to become the AC that does not divide at all (Hirsh et al, 1976;Kimble and Hirsh, 1979). In early L2, the fate of neither the Z1.ppp nor the Z4.aaa cell has been determined and neither has moved to the central position on the ventral surface of the gonad.…”

Section: Resultsmentioning

confidence: 99%

“…The VU cells undergo four rounds of cell division (Kimble and Hirsh, 1979;, whereas the AC remains a single cell in the gonad and expresses LIN-3, a member of the epidermal growth factor (EGF) family, that induces patterned proliferation of vulval precursor cells (VPCs) (Hill and Sternberg, 1992). The AC also invades the vulval epithelium by extending a process toward the 1° lineage (P6.p) of vulval cells (Sherwood and Sternberg, 2003).…”

Section: Introductionmentioning

confidence: 99%

LIN-12/Notch Regulates lag-1 and lin-12 Expression during Anchor Cell/Ventral Uterine Precursor Cell Fate Specification

Park

Choi

Hwang

2013

Molecules and Cells

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During Caenorhabditis elegans gonadal development, a stochastic interaction between the LIN-12/Notch receptor and the LAG-2/Delta ligand initiates cell fate specification of two equivalent pre-anchor cell (AC)/pre-ventral uterine (VU) precursor cells. Both cells express lin-12 and lag-2 before specification, and a small difference in LIN-12 activity leads to the exclusive expression of lin-12 in VUs and lag-2 in the AC through an unknown feedback mechanism. In this Notch signaling process, the cleaved LIN-12/Notch intracellular domain (NICD) binds to the LAG-1/CSL transcriptional repressor, forming a transcriptional activator complex containing LAG-1 and NICD. Here we show that clustered LAG-1 binding sites in lin-12 and lag-1 are involved in regulating lin-12 and lag-1 expression during AC/VU cell fate specification. Both genes are expressed in VU cells, but not the AC, after specification. We also show that lin-12 is necessary for lag-1 expression in VU cells. Interestingly, lin-12 (null) animals express lag-1 in the AC, suggesting that LIN-12 signaling is necessary for the suppression of lag-1 expression in the AC. Ectopic expression of lag-1 cDNA in the AC causes a defect in the vulvaluterine (V-U) connection; therefore, LAG-1 should be eliminated in the AC to form a normal V-U connection at a later developmental stage in wild-type animals.

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“…The answer is that, in contrast to the predominantly invariant and fully described cell lineage of the C. elegans soma (Sulston and Horvitz, 1977;Kimble and Hirsh, 1979;Sulston et al, 1983), germ cell divisions are variable with respect to order and division plane (Kimble and Hirsh, 1979). Thus, the complete cell lineage of C. elegans that has permitted unprecedented single-cell resolution correlation of genetic and anatomical studies in this organism is not applicable to the germ line.…”

Section: What Is the Evidence For C Elegans Germline Stem Cells?mentioning

confidence: 99%

“…Taken together with the data from Maciejowski et al (2006), at the lower estimate of a 16 hr cell cycle, these data suggest that metaphase/anaphase would average over 9 min and the time of phospho-histone H3-positive would be on the order of ϳ30 min. The estimated adult germ cell cycle time is also long compared with the postembryonic somatic cell divisions, which rarely approach 20 hr and are most often less than 10 hr in duration (Sulston and Horvitz, 1977;Kimble and Hirsh, 1979).…”

Section: Average Cell Cycle Time In the Adult Germ Line Is Relativelymentioning

confidence: 99%

Caenorhabditis elegans germ line: A model for stem cell biology

Hubbard

2007

Developmental Dynamics

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The postembryonic cell lineages of the hermaphrodite and male gonads in Caenorhabditis elegans

Cited by 802 publications

References 15 publications

A “FLP-Out” System for Controlled Gene Expression in Caenorhabditis elegans

A “FLP-Out” System for Controlled Gene Expression in Caenorhabditis elegans

LIN-12/Notch Regulates lag-1 and lin-12 Expression during Anchor Cell/Ventral Uterine Precursor Cell Fate Specification

Caenorhabditis elegans germ line: A model for stem cell biology

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