IngentaConnect Inhibition of MAP kinase activity moderates changes in expression...

Authors: Kojima, Takashi¹; Yamamoto, Toshinobu²; Lan, Mengdong²; Murata, Masaki²; Takano, Ken-ichi²; Go, Mitsuru²; Ichimiya, Shingo²; Chiba, Hideki²; Sawada, Norimasa²

Source: Medical Electron Microscopy, Volume 37, Number 2, June 2004 , pp. 101-113(13)

Publisher: Springer

Abstract:

The signal transduction pathways and activation of the MAP kinase or PI3 kinase signaling cascade regulate a variety of cellular processes, including proliferation and differentiation in hepatocytes. To elucidate the mechanisms of signal transmission required for the regulation of gap and tight junctions during DNA synthesis in rat hepatocytes, we determined changes of expression and function of gap and tight junctions of cells grown in primary culture, using inhibitors of signaling pathways for MAP kinase (PD98059) and PI3 kinase (LY294002). During the stimulation of DNA synthesis induced by epidermal growth factor (EGF), immunoreactivity and mRNAs of gap junction protein Cx32 and of tight junction protein claudin-1 markedly decreased with reduction of gap junctional intercellular communication (GJIC) and the fence function of tight junctions. In Western blots, whole-cell lysate of claudin-1 protein decreased and phosphorylated Cx32 protein in the insoluble fraction of Triton X-100 increased during the stimulation of DNA synthesis. During reinhibition of DNA synthesis, the changes of Cx32 and claudin-1 returned to control levels, as did both functions. In treatment with the inhibitors before DNA synthesis, PD98059 inhibited the changes of expression and function of Cx32, but not claudin-1, without inhibition of cell growth, whereas LY294002 completely inhibited cell growth. These findings indicate that the PI3 kinase pathway rather than the MAP kinase pathway plays an important role for EGF-induced proliferation of rat hepatocytes, and that changes of Cx32 in hepatocytes during the stimulation of DNA synthesis may be in part controlled through MAP kinase. Furthermore, Cx32, but not claudin-1, protein may be a target of activated MAP kinase in hepatocytes.

Keywords: Gap junctions; Tight junctions; Hepatocytes; MAPK; PI3K

Document Type: Research article

DOI: 10.1007/s00795-003-0239-7

Affiliations: 1: Department of Pathology, Sapporo Medical University School of Medicine, S1, W17, Sapporo, 060-8556, Japan, Email: ktakashi@sapmed.ac.jp 2: Department of Pathology, Sapporo Medical University School of Medicine, S1, W17, Sapporo, 060-8556, Japan,

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Inhibition of MAP kinase activity moderates changes in expression and function of Cx32 but not claudin-1 during DNA synthesis in primary cultures of rat hepatocytes