Multiple Hox/HOM-class homeoboxes in Platyhelminthes.

The importance of the Hox/HOM class of homeobox genes in early anterior-posterior pattern formation and the conserved genomic organization of this gene family provides an interesting study in genome evolution. The Platyhelminthes (flatworms) are a basal metazoan group with a simple bilateral body plan. We used the polymerase chain reaction (PCR) to detect Hox/HOM-class homeobox genes from species representing two classes of flatworms. Seven planarian and five trematode Hox/HOM-class homeoboxes were found. The sequences of the genes are consistent with the presence of one Hox/HOM-type cluster in the flatworms. Further analysis of this putative cluster may be expected to provide outgroup information for studying the evolution of the Hox/HOM clusters in the higher metazoa.

Increased assembly of clathrin occurs in response to mitogenic activation of murine lymphocytes.

The unassembled (soluble) and assembled (particulate) pools of clathrin in murine lymphocytes have been separated by centrifugation, and specifically quantified by immunoblotting of cellular extracts with an anticlathrin heavy chain monoclonal antibody. In resting spleen lymphocytes only 25-30% of the total cellular clathrin was found to be present in an assembled form. Upon activation of lymphocytes with B or T cell mitogens (lipopolysaccharide or concanavalin A), the levels of assembled clathrin increased to 60% of the total. These changes in the levels of assembled clathrin were not due to an increase in total cellular clathrin concentration following lymphocyte activation, but rather to changes in the steady state ratio of assembled to unassembled clathrin. The increase in assembled clathrin preceded the expression of transferrin receptors, as measured by the cell surface binding of an antitransferrin receptor monoclonal antibody, and maximal DNA synthesis, indicating that clathrin assembly occurs early after lymphocyte activation and precedes cell division. Immunofluorescence analysis of activated lymphocytes with an anti-clathrin heavy chain monoclonal antibody revealed a punctuate staining pattern characteristic of coated pits and vesicles. Activated B lymphocytes displayed particularly prominent staining in the perinuclear region compared to T cells, suggesting that clathrin assembly may be important for B cell functions such as immunoglobulin synthesis or secretion. These results suggest that in lymphocytes, clathrin assembly is a dynamic process that is triggered by mitogenic stimuli.