| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
|
|---|
| Research Abstract |
I investigated the characteristics and mRNA expression of hepatocyte growth factor (HGF) in the adult female monkey submandibular gland. HGF was extracted with PBS containing protease inhibitors, and purified partially by heparin affinity chromatography. With Western blot analysis, the HGF fraction showed an immunopositive protein band of 69 kDa corresponding to the a-subunit of HGF.The gene expression for HGF and its high affinity receptor, c-Met, were revealed in the monkey submandibular gland using RT-PCR, and the PCR products showed high homology to cDNA for human. Furthermore, in situ RT-PCR technique demonstrated that both the mRNA signal for HGF and c-Met were localized exclusively to the striated and excretory ductal cells. Hence, these results indicate that both HGF and c-Met are synthesized in the same cells in monkey submandibular glands.
Moreover, I investigated the characteristics and he mRNA expression of monkey submandibular gland heparan sulfate proteoglycan (HSPG), which is a low affinity receptor of HGF.PBS-soluble HSPG and microsomal membrane HSPG were extracted from submandibular glands, and purified partially by ion exchange chromatography. Western blot analysis demonstrated several PBS-soluble HSPGs and only a microsomal membrane HSPG.The RT-PCR technique revealed the mRNA expression of syndecan-1, syndecan-2, syndecan-4 and glypican-1, which are cell-surface HSPGs. These results indicated that the cell-surface HSPGs are synthesized in the monkey submandibular glands, and that their ectodomains are released into the extracellular matrix.
|
