Figure 2

Terminal end bud (TEB) and duct morphology. (a) High-magnification carmine alum-stained wholemount of a primary duct that has recently passed the central lymph node (upper left corner). The bifurcating TEB is in the final stages of forming two new primary ducts with independent TEBs. Three newly formed lateral (secondary) side-branches are also present along the trailing duct (open arrowhead), as is an area of increased cellularity that may represent a nascent lateral bud (filled arrowhead). Increased stromal cellularity is also apparent about the bifurcating TEB. Scale bar, 200 μm. (b) Immunophotomicrograph of a TEB illustrating its considerable proliferative activity, as indicated by the large number of cells that have undergone DNA replication and have thus incorporated bromodeoxyuridine (brown diaminobenzidine-stained nuclei) during a 2-hour chase period. Rather than pulling themselves forward, TEBs seem to be pushed through the adipose-rich stroma by virtue of this high proliferative activity [6]. Hematoxylin counterstaining also reveals the stromal collar, rich in fibroblasts and collagen, that characteristically surrounds the TEB neck (arrow) and its conspicuous absence beyond the invading distal cap. Scale bar, 100 μm. (c) Schematic diagram depicting the salient architectural features of TEBs and their subtending ducts, including their fibroblast-rich stromal collar and high mitotic index. Though there is no evidence that normal ductal cells ever cross the basal lamina, thinning of the basement membrane (dotted lines) does seem to occur at the tips of invading ducts as a result of their partial enzymatic degradation and/or incomplete de novo synthesis. Stromal macrophages and eosinophils are also depicted.