Aldosterone signaling modifies capillary formation by human bone marrow endothelial cells.
We investigated the regulation of the epithelial sodium channel (ENaC) in human bone marrow endothelial cells (HBMEC) responding to mineralocorticoid hormones and other accessory effectors. The message for both the mineralocorticoid receptor (MCR) and the alpha subunit of ENaC was expressed in HBMEC as predicted bands of 838 and 521 bp, respectively. In Western blots, the MCR of about 107 kDa was localized primarily in the cytoplasmic compartment but migrated to the nucleus when cell cultures were exposed to exogenous aldosterone. On the other hand, the alphaENaC was revealed as a membrane-bound protein of approximately 82 kDa, whose abundance increased after aldosterone treatment. Confocal microscopy confirmed the presence of both the MCR and ENaC as nucleocytoplasmic and membrane-bound proteins, respectively, and both colocalized with tubulin in situ. On Matrigel, the mineralocorticoid aldosterone, by itself, did not influence capillary formation by HBMEC, but the diuretic amiloride reduced the organization of HBMEC into capillary-like networks; curiously, aldosterone further exacerbated this inhibitory effect of amiloride. On the fibrin matrix, aldosterone had no influence at all on the length of the newly formed capillaries, but the capillary diameter was highly increased over the control. Aldosterone-mediated capillary swelling was totally reversed by amiloride, which, by itself, also inhibited capillary elongation by HBMEC. Thus, cell signaling by mineralocorticoid hormones in HBMEC appears to proceed in a manner very similar to that in the epithelial cell, thereby leading to an increase in the endothelial cell volume, which may underline the hypertensive state and which may also modify angiogenesis.
Chen
W
W
Inserm and Cnrs, Paris, France.
Valamanesh
F
F
Mirshahi
T
T
Soria
J
J
Tang
R
R
Agarwal
M K
MK
Mirshahi
M
M
D000450:Aldosterone / Q000494:pharmacology* / Q000502:physiology; D000584:Amiloride / Q000494:pharmacology; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002196:Capillaries / Q000187:drug effects* / Q000502:physiology; D002460:Cell Line; D004232:Diuretics / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D021122:Protein Subunits / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D018161:Receptors, Mineralocorticoid / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects*; D015222:Sodium Channels / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D014404:Tubulin / Q000378:metabolism
D004232:Diuretics; D021122:Protein Subunits; D018161:Receptors, Mineralocorticoid; D015222:Sodium Channels; D014404:Tubulin; D000450:Aldosterone; D000584:Amiloride