Immuno-gold-labelling of CUT-1, CUT-2 and cuticlin epitopes in Caenorhabditis elegans and Heterorhabditis sp. processed by high pressure freezing and freeze-substitution.
CUT-1 and CUT-2 are two distinct protein components of cuticlin, the insoluble residue of the cuticles of nematodes. In previous experiments of gold-immuno-labelling on sections of chemically fixed Caenorhabditis elegans, CUT-1 and CUT-2 epitopes were specifically lost. Cryo-immobilization of C. elegans under high pressure followed by freeze-substitution, however, resulted in a good preservation of these antigenic sites and of the ultrastructure of the worms. The entomopathogenic nematode Heterorhabditis sp. processed by the same cryopreparation protocol has shown a strong reactivity with anti-sera raised against CUT-1, CUT-2 and against the whole cuticlin residue of C. elegans. The localization of these epitopes was conserved across the two species.
Favre
R
R
International Institute of Genetics and Biophysics, CNR, Naples, Italy.
Hermann
R
R
Cermola
M
M
Hohenberg
H
H
Müller
M
M
Bazzicalupo
P
P
D000818:Animals; D000947:Antigens, Helminth / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000378:metabolism* / Q000648:ultrastructure*; D029742:Caenorhabditis elegans Proteins; D000939:Epitopes / Q000378:metabolism; D005629:Frozen Sections; D015801:Helminth Proteins / Q000276:immunology / Q000378:metabolism*; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D017172:Rhabditoidea / Q000378:metabolism* / Q000648:ultrastructure*; D013045:Species Specificity
D000947:Antigens, Helminth; C089489:CUT-2 protein, C elegans; D029742:Caenorhabditis elegans Proteins; C066374:EPICUT-1 protein, Ascaris suum; D000939:Epitopes; D015801:Helminth Proteins; C070207:CUT-1 protein, C elegans