Single injection into the cerebrospinal fluid of antibodies against the secretory material of the subcommissural organ reversibly blocks formation of Reissner's fiber: immunocytochemical investigations in the rat

Abstract

An antibody (cf. Rodríguez et al. 1984b) raised in rabbits against the glycoproteins of the bovine Reissner's fiber (RF) was injected into the lateral brain ventricle of 38 rats with the aim to interfere with RF formation. The rats were killed 20 min; 1, 4, 8, 12 h; and 1, 2, 3, 5, and 8 days after the injection. Based on the fact that the material secreted by the subcommissural organ (SCO) into the cerebrospinal fluid (CSF) first condenses on the organ surface as a distinct layer (pre-RF material) and then becomes assembled to form RF and that both structures are distinguishable in tissue sections, three immunostaining procedures were applied. They served to visualize: (i) secretory material that had not bound the injected antibody; (ii) secretory material-antibody complexes formed in vivo; and (iii) antibody not bound to its antigen and present in the ventricles and the subarachnoid space. After a single injection of the above-mentioned antibody the following events were observed: (1) The antibody was present in the brain cavities for at least 8 h. (2) The injected antibody bound selectively to the pre-RF and RF. (3) Pre-RF displayed antibody binding during the 24 h following the injection. During the 2nd and 3rd post-injection days, the pre-RF was free of antibody, indicating that it was formed by newly released secretory material. (4) Approximately 4 h after the injection, the RF detached from the SCO and underwent fragmentation. Clusters of these fragments were found in the Sylvian aqueduct and fourth ventricle. (5) In the fragmented original RF the injected antibody against Reissner's fiber remained bound throughout the entire period of observation, i.e. for 8 days. (6) In rats of the 1-, 3-, 5- and 8-day-groups, RF was missing from the central canal of the spinal cord. (7) One day after the injection, a new RF structure started to grow from the rostral end of the SCO. This newly formed fiber could be distinguished from the original RF because of (i) its normal appearance; (ii) it did not display binding of the injected antibody. (8) At day 3, the growing RF had not yet extended to the Sylvian aqueduct. (9) At day 8, the new RF reached the fourth ventricle. Control experiments involved the intraventricular administration of (i) an antibody against the secretory material extracted from the entire bovine SCO; (ii) antivasopressin; and (iii) rabbit IgG. From these only antibody (i) bound to pre-RF and RF.