Effect of papain digested, univalent antibody on the morphology, cleavage and fertilizing capacity of sea urchin eggs

• Published in: Experimental cell research Vol. 45; no. 2; pp. 433 - 449
• Main Authors: Metz, Ch.B., Thompson, Pamela H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.02.1967
• Subjects: Animals; Antigen-Antibody Reactions; Cell Division - drug effects; Coombs Test; Echinodermata - drug effects; Echinodermata - embryology; Fertilization - drug effects; gamma-Globulins - pharmacology; Immunoglobulin G - pharmacology; Papain - pharmacology; Rabbits
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/0014-4827(67)90192-9

1. 1. Antisera were prepared by immunizing rabbits with egg jelly and jellyless egg homogenates of the sea urchin, Lytechinus variegatus. Samples of the γ-globulin fraction of the antisera were degraded to the non-agglutinating, non-precipitating, univalent (3.5S) form by papain digestion and reduction. 2. 2. Most of the univalent (3.5S) antibody preparations failed to produce the morphological changes in eggs that characteristically result from treatment with multivalent (7S) antibody. These changes include egg jelly precipitation by anti-egg jelly antibody and egg wrinkling produced by anti-egg homogenate antibody. Univalent anti-egg jelly sera produced a hyaline surface effect in most egg batches and membrane elevation in a few others. 3. 3. Univalent antibody pretreated eggs failed to undergo the characteristic morphological changes upon subsequent addition of multivalent antibody. Such inhibition was complete in the case of egg surface wrinkling and partial for egg jelly precipitation. 4. 4. Univalent antibody pretreated dejellied eggs showed surface wrinkling following treatment with anti-rabbit γ-globulin sheep serum (antiglobulin or Coombs' test). It is concluded that the egg wrinkling produced by anti-egg homogenate sera depends upon the cross-linking of antigens by multivalent antibody. 5. 5. Univalent anti-egg jelly γ-globulin inhibited cleavage of fertilized eggs. However, this inhibiting action was reduced in titer as compared to the parent multivalent antibody. 6. 6. Univalent anti-egg homogenate antibody failed to affect the fertilizability of dejellied or demembranated (protease treated) eggs. 7. 7. Dejellied eggs failed to fertilize if first treated with univalent anti-egg homogenate antibody and subsequently exposed to anti-rabbit γ-globulin sheep serum (antiglobulin or Coombs' test). 8. 8. Dejellied eggs which have been rendered unfertilizable by treatment with normal, multivalent anti-egg homogenate serum recover considerable fertilizability following treatment with protease. 9. 9. It is concluded that in Lytechinus variegatus the fertilization inhibiting action of normal multivalent, 7S, anti-egg homogenate antibody depends upon cross-linking of neighboring antigens, not to the blocking of specific antigenic sites by complementary antibody.