Abstract
BackgroundDuring 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.ResultsWe 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.ConclusionsThis 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.
Highlights
During the fourth larval (L4) stage, vulval cells of C. elegans undergo extensive morphogenesis accompanied by changes in gene expression
For over 30 years, the vulval development of C. elegans has been an important model in which to study mechanisms underlying the development of complex organisms [1, 2]
Bed-3 was recently discovered to be regulated by blmp-1, another component of the heterochronic pathway [24]. These results suggest that the timing mechanism operating throughout the entire body of the worm feeds into vulval development at specific time points, allowing for precise temporal control of gene expression
Summary
During the fourth larval (L4) stage, vulval cells of C. elegans undergo extensive morphogenesis accompanied by changes in gene expression. During the fourth larval (L4) stage, these cells undergo a complex series of morphogenetic events accompanied by dynamic changes in gene expression patterns This makes it a potentially powerful system in which to study gene regulation during terminal differentiation and mechanisms that underlie complex morphogenetic processes [4,5,6,7,8,9,10]. Vulval cells migrate from where they were generated (near the original positions of vulval precursor cells induced in the L3 stage) toward the center of the future vulva These cells extend processes and fuse with one another such that they form a dorsal/ventral stack of seven toroids, called vulF, vulE, vulD, vulC, vulB2, vulB1 and vulA. Additional cell-cell connections are made; vulC and vulD make connections to the vulval muscle cells that open the vulva during egg laying, vulE makes a structural connection to lateral hypodermal
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