Aggregates of acetylcholinesterase induced by acetylcholine receptor-aggregating factor

Basal lamina-rich extracts of Torpedo californica electric organ contain a factor that causes acetylcholine receptors (AChRs) on cultured myotubes to aggregate into patches. Our previous studies have indicated that the active component of these extracts is similar to the molecules in the basal lamina which direct the aggregation of AChRs in the muscle fibre plasma membrane at regenerating neuromuscular junctions in vivo. Because it can be obtained in large amounts and assayed in controlled conditions in cell culture, the AChR-aggregating factor from electric organ may be especially useful for examining in detail how the postsynaptic apparatus of regenerating muscle is assembled. Here we demonstrate that the electric organ factor causes not only the formation of AChR aggregates on cultured myotubes, but also the formation of patches of acetylcholinesterase (AChE). This finding, together with the observation that basal lamina directs the formation of both AChR and AChE aggregates at regenerating neuromuscular junctions in vivo, leads us to hypothesize that a single component of the synaptic basal lamina causes the formation of both these synaptic specializations on regenerating myofibres.     

Identification of angiogenic activity and the cloning and expression of platelet-derived endothelial cell growth factor

Cloning and sequencing of the complementary DNA for platelet-derived endothelial cell growth factor indicates that it is a novel factor distinct from previously characterized proteins. The factor, a protein with a relative molecular mass of about 45,000, stimulates endothelial cell growth and chemotaxis in vitro and angiogenesis in vivo.     

Selective activation of Lyt 2+ precursor T cells by ligation of the antigen receptor

Resting T lymphocytes may be activated either physiologically, by the specific recognition of antigen in association with molecules encoded by the major histocompatibility complex (MHC), or non-physiologically using mitogens such as concanavalin A (Con A). The former activation process is difficult to analyse because resting precursor T cells specific for a particular antigen-MHC combination can only be isolated in the presence of a large excess of bystander cells of irrelevant specificity; clonal populations of uniform specificity are not useful for studying the activation of naive T cells because there is no reason to believe that such cloned cells ever return to the state of resting precursors. Mitogens may activate a large fraction of resting T cells, but analysis is again complicated because the target molecule(s) of most mitogens is unknown and the relationship of this kind of activation to physiological induction by antigen plus MHC molecules remains unclear. By using a monoclonal antibody specific for the antigen receptors on approximately 25% of all T cells of both Lyt 2+ and Lyt 2- subsets, we have studied the induction of lymphokine responsiveness in resting normal T cells. This antibody, immobilized on Sepharose beads, is sufficient to activate Lyt 2+ T cells, but not Lyt 2- T cells, to clonal expansion in the presence of a mixture of lymphokines (10% rat spleen Con A supernatant). We report here that clonal growth of the T cells obeys single-hit kinetics in limiting-dilution microcultures, suggesting that a single cell type is limiting. We conclude that cytotoxic T-lymphocyte (Tc) precursors require only ligation of the antigen receptor before they become responsive to lymphokines, whereas helper T-lymphocyte (Th) precursors require additional signals.     

Isolation and characterization of hemagglutinins from the sponge Dysidea herbacea

The sponge Dysidea herbacea (Keller) was found to possess hemagglutinins. The major component, DHA-I, is a protein with a mol. wt of 26,000, which dissociates into subunits of equal size (14,000). It contains large amounts of glutamic acid and aspartic acid residues, but no half-cystine, methionine or histidine residues. DHA-I reacted with rabbit and human AB0 erythrocytes. D-galactose and lactose were effective inhibitors of DHA-I. The sponge also contained a minor component(s) which reacted preferentially with rabbit erythrocytes but not with human AB0 erythrocytes.     

Differences in cardiac myosin light chain LC1 among human, monkey and sheep

The atrial and ventricular myosin light chains of human, monkey and sheep hearts were compared by dodecylsulfate polyacrylamide gel electrophoresis. The atrial light chain 2 and ventricular light chain 2 are similar among these mammals. However, the atrial light chain 1 of monkey has different electrophoretic mobility from those of human and sheep. The monkey ventricular light chain 1 has same mobility as that of sheep but different from that of human.     

Light-dependent antibody labelling of photoreceptors

Monoclonal antibodies are tools widely used to analyse the structure of the nervous system. Whereas some labelling patterns are highly reproducible, others appear to vary from one preparation to the next, as we noticed in particular for some antibodies with respect to photoreceptor labelling. To establish whether some of this variability can be linked to functional criteria, we tested for light-dependence. We found two antibodies that label photoreceptor outer segments, only when the retina has been illuminated, and a third antibody that has a selective affinity for dark-adapted outer segments. The two antibodies against light-activated sites are primarily directed against the highly phosphorylated neurofilament subunit at relative molecular mass 200,000 (200K). One of them, RT97, recognizes on immunoblots, in addition to neurofilaments, a light-activated epitope on a protein that resembles the photopigment rhodopsin, presumably a phosphorylation-dependent site. Antibodies like those described here may allow the study of physiological processes such as light and dark adaptation using morphological techniques.