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Neuropeptide Y expression, localization and cellular transducing effects in HUVEC
Author(s) -
Silva Antonio P.,
Kaufmann Jocelyne E.,
Vivancos Cécile,
Fakan Stanislav,
Cavadas Claudia,
Shaw Phillip,
Brunner Hans R.,
Vischer Ulrich,
Grouzmann Eric
Publication year - 2005
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1042/bc20040102
Subject(s) - neuropeptide y receptor , biology , histamine , medicine , receptor , endocrinology , autocrine signalling , umbilical vein , immunofluorescence , microbiology and biotechnology , neuropeptide , biochemistry , immunology , in vitro , antibody
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/μg of protein and 0.46 fmol of pro‐NPY/μg 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.