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.
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.