Abstract

We describe a novel screen to isolate pharyngeal cell morphology mutants in Caenorhabditis elegans using myo-2::GFP to rapidly identify abnormally shaped pharynxes in EMS (Ethyl Methanesulfonate) mutagenized worms. We observed over 83 C. elegans lines with distinctive pharyngeal phenotypes in worms surviving to the L1 larval stage, with phenotypes ranging from short pharynx, unattached pharynx, missing cells, asymmetric morphology, and non-adherent pharynx cells. Thirteen of these mutations have been chromosomally mapped using Single Nucleotide Polymorphisms (SNPs) and deficiency strain complementation. Our studies have focused on genetically mapping and functionally testing two phenotypes, the short pharynx and the loss of muscle cohesion phenotypes. We have also identified new alleles of sma-1, and our screen suggests many genes directing pharynx assembly and structure may be either pharynx specific or less critical in other tissues.

Highlights

  • The C. elegans pharynx exhibits progressive restriction of cell fate during development, resulting in the expression of differentiation factors and structural proteins essential to its function as a neuromuscular pump [1,2,3]

  • The initial goal of this project was to determine the genes involved in posterior pharyngeal muscle fate

  • Previous studies have shown that the transcription factor, TBX-2, mediates anterior muscle fate

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Summary

Introduction

The C. elegans pharynx exhibits progressive restriction of cell fate during development, resulting in the expression of differentiation factors and structural proteins essential to its function as a neuromuscular pump [1,2,3]. In C. elegans, pha-4 is an organ identity gene involved in the specification and differentiation of all cells destined to become the pharynx [5,6,7]. Mutations in glp-1, tbx-35, or lag-1 result in a loss of all pharynx cells derived from ABa or MS lineage, resulting in formation of a half pharynx. In the cases of pha-4, glp-1, tbx-35, and lag-1, the loss of pharynx cells is not cell-type specific, rather entire regions of the pharynx are deleted such as the anterior ABa derived-pharynx in glp-1 mutants [5,8,10,11,12,13]

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