SpringerProtocols: Abstract: Quantitative Analysis of Distal Tip Cell Migration in C. elegans

8. Quantitative Analysis of Distal Tip Cell Migration in C. elegans

By: Myeongwoo Lee¹, Erin J. Cram²

Abstract

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Correct distal tip cell (DTC) migration in the nematode C. elegans requires sensing soluble and matrix cues, remodeling extracellular matrix, and signaling through conserved integrin and netrin pathways. The DTC executes a complex path and coordinates its migration with the developmental stages of larval morphogenesis. This chapter outlines a method for investigating DTC migration in C. elegans using feeding RNA interference (RNAi) and light microscopy. To deplete a candidate gene of interest, nematode eggs are added to plates seeded with RNAi-inducing bacterial lawns. The animals hatch and begin to eat the RNAi bacteria, releasing dsRNA and causing the targeted gene to be depleted during larval development. Positions of migratory cells are monitored in larvae and young adults using differential interference contrast (DIC) and epifluorescence microscopy.

Affiliation(s): (1) Department of Biology, Baylor University, Waco, TX, USA
(2) Department of Biology, Northeastern University, Boston, MA, USA

Book Title: Chemotaxis: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 571 | Pub. Date: Sep-08-2009 | Page Range: 125-136 | DOI: 10.1007/978-1-60761-198-1_8

Subject: Cell Biology

Key Words: DTC - Cell migration - C. elegans - RNAi - DIC microscopy - Fluorescence microscopy

References

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