The transport of exogenous peroxidase across the blood-tissue-lymph interface1

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Horseradish peroxidase has been used as an electron opaque probe to trace movement of this protein across the blood-tissue-lymph interface at the electron microscopic level. Peroxidase was injected into the tail veins of mice and localized by the cytochemical procedures of Graham and Karnovsky within blood capillaries, connective tissue, and lymphatics in the dermis of mouse ears. This procedure provides the advantage of investigating the transport of a protein across the blood-tissue-lymph interface without the increase of fluid pressure that is possible with microinjection procedures. Within 1 minute peroxidase staining is observed in the blood capillaries, throughout the connective tissue areas between collagen fibrils, and within macrophages. A strong reaction product is observed along the endothelial plasma membranes and within the lymphatic lumen. Also at the early time periods (1–5 minutes), reaction product is observed in plasmalemmal vesicles which open onto both connective tissue and luminal surfaces of the endothelium, as well as in vesicles which appear to be in transit through the cytoplasm. Enzyme staining is also prominent throughout the clefts of intercellular junctions. The reaction product within the lymphatic lumen reaches its maximal concentration between 15 and 30 minutes; thereafter its staining intensity declines markedly so that by 24 hours very little staining is observed in either the interstitium or the lymphatic lumen. However, peroxidase staining is observed in macrophages and vesicles of varying sizes in the lymphatic endothelium. It is concluded that clefts of intercellular junctions provide the major passageway for transendothelial movement between the interstitium and the lymphatic capillary lumen. The presence of peroxidase in vacuoles for time periods of up to 24 hours is discussed in relation to previous findings, which suggest that the accretion of proteins and tracers within lysosomal vacuoles is indicative of an intracellular digestion of engulfed substances for release from the lymphatic endothelium.

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      Citation Excerpt :

      Other than intercellular pathways, there is ample evidence for transendothelial pathways for solute and lipid transport in the initial lymphatics. Leak has used a number of solutes such as horseradish peroxidase, ferritin, carbon, and latex microspheres to demonstrate plasmalemmal vesicles in the skin [19,64,68,118]. This pathway may be especially important in organs where open junctions in the initial lymphatics are seldom seen, such as the lung [119], intestine [120], and diaphragm [121].

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    1

    This work was supported by USPHS Grant Number AI-07348 and the Shriners Burns Institute, Boston Unit, and USPHS Grant Number HE-13901.

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