Worm Breeder's Gazette 2(2): 11

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Biochemical Studies of C. elegans Cuticles

M. Kusch, J. Laufer, R. Edgar

Cuticles from C.  elegans have been isolated by sonication of adult 
nematodes in tris buffer (10 mM, pH 7.6) containing N-ethyl maleimide (
7.5 mM) and EDTA (10 mM).  Soluble proteins were removed by washing 
and cuticles further extracted by incubation in either sodium chloride 
(0.6 - 1.0 M) on ice or SDS (1%) at room temperature.  Phase contrast 
microscopy of such cuticles reveals a structure with inner and outer 
refractile layers and with regularly spaced refractile dots along the 
annulae.  The dots may correspond to  struts  between the two layers 
of cuticle.  The inner cuticle layer and the struts are solubilized by 
incubation of cuticles in SDS (1%) containing  -mercaptoethanol (1%).  
SDS polyacrylamide gel electrophoresis of  -mercaptoethanol-released 
material reveals several bands of high molecular weight material.  The 
inner layer and struts are sensitive to digestion by trypsin and 
collagenase whereas papain appears to digest only struts.  Cuticles 
are not visibly affected by chitinase or lipase although chitinase 
appears to digest broken egg  shells .  Attempts to solubilize the 
outer cuticle layer with SDS, urea or guanidine hydrochloride have 
been unsuccessful.  Work is now in progress to characterize cuticular 
components using biochemical and immunological techniques.