Keiichi Ozaki

A monoclonal antibody directed to Tn antigen

A murine monoclonal antibody, MLS 128, that was assigned to an anti-Tn antibody has been established by immunizing mice with human colonic cancer cells (LS 180). MLS 128 bound to mucin glycopeptides from LS 180 cells and their asialo forms to the same extent as well as to ovine submaxillary mucin (OSM) and asialo OSM. Special non-sialylated GalNAc residue(s) attached to a certain peptide region in the antigens seems to be involved in the binding since N-acetylgalactosaminidase treatment of the antigen abolished the binding and pronase digestion diminished the binding markedly.

Localization of fibroblast growth factor-9 mRNA in the rat brain

We examined the localization of fibroblast growth factor-9 (FGF-9) mRNA in the rat brain by in situ hybridization. FGF-9 mRNA was moderately or weakly expressed in widespread regions including the olfactory bulb, caudate putamen, cerebral cortex, hippocampus, thalamus, hypothalamus, midbrain, brainstem and cerebellum. However, FGF-9 mRNA was also strongly expressed in several specific nuclei including the red nucleus and oculomotor nucleus in the midbrain, the vestibular nucleus and facial nucleus in the brainstem and the medial cerebellar nucleus, interposed cerebellar nucleus and lateral cerebellar nucleus in the cerebellum. The cellular localization of FGF-9 mRNA indicated that the mRNA in the rat brain was expressed preferentially in neurons, although FGF-9 was originally isolated from human glioma cells. The localization profile of FGF-9 mRNA is different from those of aFGF, bFGF and FGF-5 mRNAs reported previously. The present findings indicate that FGF-9 has a unique role in the brain.

The differences in structural specificity for recognition and binding between asialoglycoprotein receptors of liver and macrophages

The Gal/GalNAc-specific lectin on the surface of rat peritoneal macrophages (macrophage asialoglycoprotein binding protein, M-ASGP-BP), which consists of a single polypeptide chain of 42 kDa, can form a homooligomeric receptor exhibiting high affinity for asialoorosomucoid (ASOR) [Ozaki K., Ii M., Itoh N., Kawasaki T. (1992)J Biol Chem267: 9229–35]. In this study, the binding affinity of M-ASGP-BP was studied by using a series of synthetic or natural glycosides as inhibitors of125I-ASOR binding to recombinant M-ASGP-BP expressed on COS-1 cells (rM-ASGP-BP), and the results were compared with those of human hepatic lectin (HHL) on Hep G2 cells. Clustering of multiple Gal (or GalNAc) residues increased the binding affinity to M-ASGP-BP as well as to HHL. In contrast to HHL and other mammalian hepatic lectins, rM-ASGP-BP bound Gal residues tighter than GalNAc residues. A galactose-terminated triantennary N-glycoside, having oneN-acetyl-lactosamine unit on the 6 branch and twoN-acetyl-lactosamine units on the 3 branch of the trimannosyl core structure, showed affinity enhancement of ∼105 over a monovalent ligand for HHL, while the same glycopeptide showed enhancement of about 2000-fold for rM-ASGP-BP. These results suggest that spatial arrangements of sugar combining sites and subunit organization of macrophage and hepatic lectins are different.

Rat FGF receptor-4 mRNA in the brain is expressed preferentially in the medial habenular nucleus

The fibroblast growth factor (FGF) receptor family consists of four members, FGFR-1, FGFR-2, FGFR-3 and FGFR-4, that are closely related receptor tyrosine kinases. We examined the expression of rat FGFR-4 mRNA in the brain by in situ hybridization and compared it with that of the mRNAs for other FGF receptors. In contrast with FGFR-1, FGFR-2 and FGFR-3 mRNAs which are expressed widely in the brain, the FGFR-4 mRNA in the brain is expressed preferentially in the medial habenular nucleus neurons. The present finding indicates that FGFR-4 has a function specific to the medial habenular nucleus.

Expression of insulin receptor-related receptor in the rat brain examined by in situ hybridization and immunohistochemistry.

Insulin receptor-related receptor (IRR) is a member of the insulin receptor family. However, its endogenous ligand and physiological roles are unknown. To elucidate the physiological roles of IRR, an orphan receptor, in the brain, we examined its expression at mRNA and protein levels in the brain by in situ hybridization and immunohistochemistry, respectively. The expression of IRR mRNA in the brain was highly restricted to the forebrain including the nucleus of the diagonal band, medial septal nucleus, ventral pallidum, accumbens nucleus and caudate putamen, and the brainstem including the prepositus hypoglossal nucleus, medial vestibular nucleus, gigantocell reticular nucleus, paragigantocellular nucleus and ventral cochlear nucleus. Most IRR mRNA-positive cells in the forebrain but not in the brainstem were cholinergic neurons. However, most IRR mRNA in the forebrain and brainstem was coexpressed with that of trkA, a high-affinity receptor for nerve growth factor. IRR-immunoreactive cell bodies were also detected in the forebrain and brainstem. The pattern of IRR immunoreactivity was similar to that of IRR mRNA. Its restricted pattern indicates that IRR plays unique roles in the brain, in contrast to insulin and insulin-like growth factor-I receptors, other members of the insulin receptor family, which are widely expressed in the brain.

Expression of insulin receptor-related receptor rnRNA in the rat brain is highly restricted to forebrain cholinergic neurons☆

Insulin receptor-related receptor (IRR) is a member of the insulin receptor family. However, its endogenous ligand and the physiological roles of IRR are unknown. To elucidate the physiological roles of IRR in the brain, we examined the expression of its mRNA in the rat brain by in situ hybridization. In contrast to the widespread expression of insulin receptor and insulin-like growth factor-1 receptor mRNAs in the brain, the expression of IRR mRNA was highly restricted to the forebrain cholinergic neurons. All the forebrain cholinergic neurons expressed IRR mRNA. The present findings indicate that IRR has a selective role in the brain for forebrain cholinergic function.