Neuropeptide Y expression, localization and cellular transducing effects in HUVEC

Details

Serval ID
serval:BIB_D08AFD113770
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Neuropeptide Y expression, localization and cellular transducing effects in HUVEC
Journal
Biologie Cellulaire
Author(s)
Silva  A. P., Kaufmann  J. E., Vivancos  C., Fakan  S., Cavadas  C., Shaw  P., Brunner  H. R., Vischer  U., Grouzmann  E.
ISSN
0248-4900 (Print)
Publication state
Published
Issued date
06/2005
Volume
97
Number
6
Pages
457-67
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jun
Abstract
BACKGROUND INFORMATION: NPY (neuropeptide Y) may have an effect on the properties of vascular endothelial cells such as pro-angiogenic effects and potentiation of noradrenaline-induced vasoconstriction. In HUVEC (human umbilical-vein endothelial cells), immunoreactive neuropeptide Y has been detected, but NPY synthesis, storage and secretion have not been studied. The aim of the present study was to establish NPY expression, storage and cellular transducing effects in HUVEC. RESULTS: HUVEC contain 0.19 fmol of NPY/microg of protein and 0.46 fmol of pro-NPY/microg of protein, as measured by ELISA. RT (reverse transcriptase)-PCR confirmed the expression of NPY in HUVEC. Immunofluorescence revealed the presence of NPY in small punctate structures, with a fluorescence pattern different from that observed for von Willebrand factor, indicating distinct storage compartments. Double labelling for NPY and Rab3A demonstrated similar granular patterns, with at least partial co-localization. Electron microscopy showed NPY immunoreactivity in vesicle-like cytoplasmic structures, of a fine fibrillar texture, as well as in mitochondria and in the nucleus. A similar general distribution pattern was also obtained for Rab3A. Y1 and Y2 receptors were expressed in HUVEC as assessed by RT-PCR, and they were functional since NPY induced a 42 nM intracellular calcium increase within 100 s, representing 22% of the histamine-induced response. In contrast with histamine, NPY did not induce acute von Willebrand factor secretion. CONCLUSIONS: HUVEC produce, store and respond to NPY, suggesting an autocrine regulatory role for NPY in the endothelium.
Keywords
Calcium/metabolism Cell Nucleus/metabolism Cells, Cultured Culture Media, Serum-Free/pharmacology Cyclic AMP/metabolism Endothelium, Vascular/*metabolism Enzyme-Linked Immunosorbent Assay Gold Colloid/pharmacology Histamine/metabolism Humans Immunohistochemistry Kinetics Microscopy, Electron Microscopy, Fluorescence Microscopy, Immunoelectron Mitochondria/metabolism Neuropeptide Y/*biosynthesis RNA, Messenger/metabolism Reverse Transcriptase Polymerase Chain Reaction Signal Transduction Umbilical Veins/*cytology rab3A GTP-Binding Protein/metabolism von Willebrand Factor/metabolism/secretion
Pubmed
Web of science
Open Access
Yes
Create date
25/01/2008 10:55
Last modification date
20/08/2019 15:50
Usage data