Phenotype of normal cutaneous microvasculature - Immunoelectron microscopic observations with emphasis on the differences between blood vessels and lymphatics

The lymphatic system has been poorly characterized in comparison to the blood vessels. We investigated the expression of microvasculature markers in cutaneous lymphatics and blood microvessels in normal skin. Scrotal skin was chosen because of its high density of both types of microvessels. A pre-embedding peroxidase-conjugated immunoelectron microscopy technique was used, allowing both the visualization of the lymph and blood vessels and their immunohistochemical staining. The markers studied included endothelial antigens (recognized by PAL-E, EN-4, and von Willebrand factor/factor VIII-related antigen), structural molecules of the vascular wall (cu-smooth muscle actin, heparan sulfate proteoglycan, collagen type IV), and adhesion molecules (endothelial leukocyte adhesion molecule-1 [E-selectin], intercellular adhesion molecule-1. [ICAM-1], platelet endothelial adhesion molecule-1 [PECAM-1], vascular cell adhesion molecule-1 [VCAM-i]). It is shown that lymphatics of normal skin are phenotypically different from blood microvasculature, only weakly expressing endothelial markers (EN-4(+), von Willebrand factor/factor VIII-related antigen(+/-), PAGE(-)), mural markers (alpha-smooth muscle actin(-), heparan sulfate proteoglycan(-), collagen type IV+) and do not express the studied adhesion molecules except PE-CAM-1 (E-selectin(-), ICAM-1(-), PECAM-1(-), VCAM-1(-)). The results were substantiated by a double-labeling immunoelectron microscopic technique, which facilitates detection and assessment of microvascular segments. By this technique, collagen type IV, recognized by a peroxidase-labeled 2nd antibody, stains the basal lamina by a linear pattern, whereas a second optional epitope is visualized as grains by a silver-enhanced ultra-small gold-conjugated antibody. Our study shows that not only morphology but also antigenic phenotype of lymphatics differs significantly from blood vessels.