Fig. 1.

Perivascular-resident cells identified as melanocyte-like macrophages are in close contact with capillaries through cytoplasmic processes. (A) 3D reconstructed confocal image of the SV. PVM/Ms are labeled with an antibody for F4/80 (white), and the cytoskeleton is labeled with phalloidin (red). The PVM/Ms are sandwiched between marginal (MC) and basal cell (BC) layers of the SV. (B) 3D reconstructed images at different angles show that PVM/Ms are situated in or under subepithelial marginal cells (Left) and have less contact with basal cells (Right). (C and D) An overview (C) and a close-up view (D) of the 3D rendering of confocal stacks show the ramified processes of PVM/Ms interfacing with the endothelial tube. Capillaries are labeled with antibody for collagen IV. (E) PVM/Ms contain melanin pigment granules (arrow). (F) Triple labeling for F4/80 (green), GSTα4 (blue), and capillaries (red) in whole-mount SV shows that PVM/Ms express GSTα4. (G) Double-labeling for F4/80 (green) and GST (red) in the whole-mount SV shows that PVM/Ms express GST. (H) Double-labeling for F4/80 (green) and Kir4.1 (red) in the whole-mount SV shows that PVM/Ms express Kir4.1. (I) mRNA for Gpf480, Gst, and Kir4.1 was detected in isolated and purified PVM/Ms by RT-PCR analysis. (J and K) Primary cultured PVM/Ms are triple-labeled for GST (red) or GST4a (red), F4/80 (green), and nuclei (blue). (L) Primary cultured PVM/Ms are double-labeled for F4/80 (green) and Kir4.1. (M) mRNA for Gpf480, Gst, Gst4α, and Kir4.1 was found in primary cultured PVM/Ms by RT-PCR analysis.