Premium
Measurement of mouse vasoactive intestinal peptide receptor – 1 in RAW 264.7 cells
Author(s) -
Nelson Ashley,
Bakke Danika,
Dorsam Glenn
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb114
Subject(s) - vasoactive intestinal peptide , receptor , microbiology and biotechnology , flow cytometry , g protein coupled receptor , biology , immune system , signal transduction , cell , neuropeptide , immunology , biochemistry
Mouse vasoactive intestinal peptide receptor – 1 (VPAC1) signaling in lymphocytes has been shown to regulate proliferation, apoptosis and differentiation. VPAC1 is expressed in numerous cell types within the nervous and immune system. Recently, our laboratory has demonstrated that VPAC1 is dramatically downregulated in activated T cells mediated by a Src/ZAP70/Jnk pathway. Based on these results, we hypothesized that this neuropeptide receptor may also be regulated in a similar fashion in macrophages dependent on its activation status. Therefore, this study asked whether functional VPAC1 protein levels changed in resting and LPS treated RAW 264.7 macrophage cells. End‐point analyses will focus on 1.) mRNA levels by RT‐qPCR, 2.) cell surface protein levels by flow cytometry, and 3.) functional coupling to G as by i[cAMP] ELISA. To date, our results have shown that there is no detectable VPAC1 protein on the surface of resting RAW cells. We expect to show that functional VPAC1 protein is upregulated at the mRNA and protein levels upon LPS exposure of RAW cells that facilitates greater G as signaling. Funding: (COBRE) 2P20RR015566