Mutant Sensory Cilia in the Nematode Caenorhabditis elegans

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Table of contents  -  Abstract  -   Introduction  -   Materials & Methods  -   Results  -   Discussion  -   References

Material and Methods

Brenner (1974) describes culturing and genetic manipulation of Caenorhabditis elegans. Strains were kindly provided by Martin Chalfie, David Dusenbery, Jonathan Hodgkin, Donald Riddle, Richard Russell, and the Caenorhabditis Genetics Center at the University of Missouri, Columbia.

Chemosensory neurons were stained with fluorescein isothiocyanate and examined by fluorescence microscopy as in Hedgecock et al (1985). New mutants with abnormal staining were induced with ethylmethanesulphonate (Brenner, 1974). The cat-6 (e1861) mutation was separated from the strain CB246.

Animals were fixed for electron microscopy using glutaraldehyde and then osmium as in Sulston et al. (1983). Usually, two or three individuals of each mutant strain were embedded and sectioned together. About 200 contiguous sections, 50 nm thick, were collected from the tips of the heads. One animal, selected for good fixation quality and orientation, was photographed approximately every third section at 5000X magnification to reconstruct the head sensilla. The other animals were examined directly in the microscope.

Adapted by Yusuf KARABEY for WORMATLAS, 2003