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Morphologically defined sub-stages of C. elegans vulval development in the fourth larval stage

Mok, Darren Z. L. and Sternberg, Paul W. and Inoue, Takao (2015) Morphologically defined sub-stages of C. elegans vulval development in the fourth larval stage. BMC Developmental Biology, 15 . Art. No. 26. ISSN 1471-213X. PMCID PMC4464634.

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Background: During the fourth larval (L4) stage, vulval cells of C. elegans undergo extensive morphogenesis accompanied by changes in gene expression. This phase of vulval development, occurring after the well-studied induction of vulval cells, is not well understood but is potentially a useful context in which to study how a complex temporal sequence of events is regulated during development. However, a system for precisely describing different phases of vulval development in the L4 stage has been lacking. Results: We defined ten sub-stages of L4 based on morphological criteria as observed using Nomarski microscopy (L4.0 ~ L4.9). Precise timing of each sub-stage at 20 °C was determined. We also re-examined the timing of expression for several gene expression markers, and correlated the sub-stages with the timing of other developmental events in the vulva and the uterus. Conclusions: This scheme allows the developmental timing of an L4 individual to be determined at approximately one-hour resolution without the need to resort to time course experiments. These well-defined developmental stages will enable more precise description of gene expression and other developmental events.

Item Type:Article
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URLURL TypeDescription CentralArticle
Sternberg, Paul W.0000-0002-7699-0173
Additional Information:© 2015 Mok et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated. Received: 18 February 2015. Accepted: 3 June 2015. Published online: 12 June 2015. We thank Goh Kah Yee for preliminary analysis of best-13:gfp expression. We thank Marie-Anne Felix for the daf-6:cfp plasmid and strain. Some strains used in this study were provided by the Caenorhabditis Genetics Center, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). This work was supported by the Singapore Ministry of Education Academic Research Fund Tier 1 grant (T13-1001-P02). PWS is an Investigator of the Howard Hughes Medical Institute. Authors’ contributions: TI and PWS developed the L4 sub-staging scheme. DZLM determined the timing of sub-stages and the expression of reporters in sub-stages. TI and DZLM determined the correlation with other developmental events. The manuscript was drafted by DZLM and edited by all authors. All authors read and approved the final manuscript.
Funding AgencyGrant Number
NIHP40 OD010440
Ministry of Education (Singapore)T13-1001-P02
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Subject Keywords:C. elegans, Vulval development, Developmental timing, Gene expression
PubMed Central ID:PMC4464634
Record Number:CaltechAUTHORS:20150710-104105215
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Official Citation:Morphologically defined sub-stages of C. elegans vulval development in the fourth larval stage Darren Mok, Paul Sternberg, Takao Inoue BMC Developmental Biology 2015, 15:26 (12 June 2015)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:58845
Deposited By: Ruth Sustaita
Deposited On:10 Jul 2015 18:09
Last Modified:05 May 2020 18:55

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