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Microarray analysis of gamma melanocyte stimulating hormone (gamma‐MSH) responsive genes in melanocortin 3‐receptor (MC‐3‐R) transfected neuronal cells
Author(s) -
HughesDarden Cleo Adelle,
Redmond Brian,
Bernard Segun,
Broadway Marquita,
Wachira James
Publication year - 2007
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.21.5.a255
Subject(s) - protein kinase c , biology , melanocortin 3 receptor , transfection , signal transduction , microarray analysis techniques , microbiology and biotechnology , melanocortin , receptor , gene , gene expression , genetics , melanocortin receptor
Melanocortins are peptide hormones derived from the precursor polypeptide pro‐opiomelanocortin. They mediate their effects through a family of five G‐protein coupled receptors, by stimulating adenylate cyclase. Studies have indicated other signaling pathways such as the PKC and MAP kinase. MC3R and MC4R were implicated in obesity, insulin resistance and salt‐sensitive hypertension through gene knockout studies. Our previous studies indicated that (−2‐MSH/MC3R functions via the PKC pathway. In order to understand the molecular mechanisms involved in MC3R signaling, we treated MC3R/GFP and GFP transfected cells with 100 nM gamma‐MSH and isolated total RNA for gene transcription analysis using oligonucleotide microarrays. The GeneSifter software identified 88 up‐regulated genes and 91 down‐regulated genes. Several signaling pathway genes were altered. Q‐PCR data indicated that PTPN13 and PKC nu/PKD3 genes were up‐regulated as a result of MC3R activation by gamma‐MSH. Preliminary immunocytochemistry analyses of PKC nu/PKD3 suggested that in MC3R transfected cells, PKC nu/PKD3 was localized to the cell periphery and treatment with gamma‐MSH resulted in cytoplasmic distribution. These data suggest that PKC nu/PKD3 may play a role in MC3R signaling. This work was supported by NIH/NCRR/RCMI grant G12RR17581‐05 and the RCMI funded Molecular and Cellular and Cytology/Histology core facilities.