Differential effects of protease digestion on photoreceptor lectin binding sites.
The use of lectin cytochemistry together with proteolytic digestion techniques to partially characterize lectin binding sites of several intracellular compartments in frog photoreceptors was studied. Uniform access of reagents to all intracellular compartments was obtained by performing the experiments directly on semithin sections of retinal tissue embedded in a hydrophilic plastic resin. Protease pretreatment of sections of Xenopus laevis eyecup leads to a loss of wheat germ agglutinin (WGA) binding sites from most of the rod outer segment. Under experimental conditions used here, cone outer segment WGA binding sites are resistant to proteolytic digestion. Another major difference between rod and cone under segments is that rod outer segments are heavily labeled with succinylated WGA, whereas cone outer segments are barely labeled except for a region of intense staining thought to be at the connecting cilium. WGA binding sites in the shed outer segment tip (phagosome) are also relatively resistant to proteolytic digestion, as is the tip region of a few rod outer segments. This difference in lectin binding properties between the bulk of the outer segment membrane and the shed outer segment membrane is the only distinction we have observed between the two compartments in terms of their glycoconjugates. These results may be useful in terms of designing experiments to isolate cone and rod outer segments separately. They indicate that a change in outer segment glycoconjugates may accompany the shedding and phagocytosis events, as previously suggested, but this change does not necessarily involve the addition of saccharides to outer segment glycoproteins.
D000818:Animals; D001665:Binding Sites; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006651:Histocytochemistry; D037102:Lectins / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000378:metabolism*; D012374:Rod Cell Outer Segment / Q000378:metabolism; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins; D014982:Xenopus laevis
D005452:Fluoresceins; D037102:Lectins; D011975:Receptors, Mitogen; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins; D010447:Peptide Hydrolases; D016650:Fluorescein-5-isothiocyanate