Responsiveness to Retinoic Acid Changes during Chondrocyte Maturation

We previously showed that retinoic acid (RA) participates in the regulation of chondrocyte maturation during endochondral ossification, a process involving multiple developmental stages. To assess whether the responsiveness to RA treatment changes during chondrocyte maturation, immature chondrocytes were isolated from the caudal portion of Day 18-19 chick embryo sterna, a portion that remains cartilaginous through early postnatal life but ossifies with age. The immature cells were allowed to reach different stages of maturation by growth for different time in culture. Progression by the cells toward the mature phenotype during culture was confirmed by increases in average cell diameter, proteoglycan synthesis, and alkaline phosphatase (APase) activity. When developmentally immature passage 0 (PO) cultures were treated with RA (10-100 nM) for 72 h, the cells readily became fibroblastic, reduced drastically their proteoglycan synthesis, and failed to activate type X collagen gene expression. When older cultures (P1 and P2) were treated with RA, the cells acquired a characteristic epithelioid shape and increased their APase activity. Moreover, 5-10% of P1 cells and 20-25% of P2 cells activated type X collagen synthesis in response to RA. RA treatment markedly induced expression of the gene encoding the β isoform of retinoic acid receptor (RARβ) and also provoked a moderate 2.5-fold increase in RARα gene expression. A similar change in responsiveness to RA was observed during maturation in vivo. Chondrocytes were isolated from the cephalic portion of Day 10, 11, 13, and 16 chick embryo sterna, and were treated with different doses of RA (10-100 nM) for 72 h. The cells from the Day 10 sternum failed to activate type X collagen gene expression in response to RA. In contrast, with increasing age of the embryos, an increasing fraction of cells induced type X collagen gene expression in response to RA. We conclude that responsiveness to RA changes during the early stages of chondrocyte maturation and that maturation depends on interactions between exogenous retinoids and the endogenous developmental program of chondrocytes.     

Role of Pax-5 in the regulation of a mid-hindbrain organizer''s activity

The mes-metencephalic boundary (isthmus) has been suggested to act as an organizer in the development of the optic tectum. Pax-5 was cloned as a candidate for regulator of the organizing center. Isthmus-specific expression of Pax-5 and analogy with the genetic cascade in Drosophila suggest that Pax-5 may be at a higher hierarchical position in the gene regulation cascade of tectum development. To examine this possibility, a gain-of-function experiment on Pax-5 was carried out. In ovo electroporation on E2 chick brain with the eucaryotic expression vector that encodes chick Pax-5 cDNA was used. Not only was a considerable amount of Pax-5 expressed ectopically in the transfected brain, but irregular bulging of the neuroepithelium was induced in the diencephalon and mesencephalon. At Pax-5 misexpressing sites, uptake of BrdU was increased. Histological examination of E7 transfected brain revealed that Pax-5 caused transdifferentiation of diencephalon into the tectum-like structure. In the bulges of the E7 mesencephalon, differentiation of laminar structure was repressed when compared to the normal side. In transfected embryos, En-2, Wnt-1 and Fgf8 were up-regulated ectopically, and Otx2 was down-regulated in the diencephalon to mesencephalon. Moreover, Ephrin-A2, which is expressed specifically in the tectum with a gradient highest at the caudal end, is suggested to be involved in pathfinding of the retinal fibers, and was induced in the bulges. When the mouse Fgf8 expression vector was electroporated, Pax-5 and chick Fgf8 were also induced ectopically. These results suggest that Pax-5, together with Fgf8, hold a higher position in the genetic hierarchy of the isthmus organizing center and regulate its activity.     

Carbohydrate structures of a normal counterpart of the carcinoembryonic antigen produced by colon epithelial cells of normal adults

Normal Faecal antigen-2 (NFA-2) and non-specific crossreactingantigen-2 (NCA-2), cross-reacting with anticarcinoembryonicantigen (CEA) antibodies, were found in normal human faececand meconium, respectively. Because NFA-2, NCA-2 and CEA areconsidered as the same gene products, NFA-2 and NCA-2 shouldbe normal counterparts of CEA produced by colon epithelial cellsof normal adults and fetuses, respectively. Comparison of sugarchain structures of these three antigens is indispensable inorder to unravel the stnaural alteration induced by malignanttransformation and development of colon epithelial cells. Thesugar chain structures of CEA (Yamashita,K. et al., Cancer Res.,47,3451–3459,1987) and NCA-2 (Yarnashita,K. et al., J.Bid Chem, 264,17873-17881,1989) were previously reported. Inthis paper, the structures of the oligosaccharides releasedfrom four NFA-2 samples by hydrazinolysis were studied by meansof lectin-affinity column chromatography, endo- and em-glycosidasedigestion, methylation analysis, hydrazinolys-nitrous acid deaminationand electrospray ionization mass spectrometry. NFA-2 contains24–27 mol of N-linked sugar chains/molecule, which issimilar to NCA-2 (27 mol) and CEA (24–27 mol). In contrastto CEA, which contains {small tilde} 8% high-mannose-type sugarchains all sugar chains of NFA-2 are mono- to tetra-antennarycomplex-type chains having four types of tri-mannosyl cores,with or without bisecting N-acetylglucosamine and fucose residues.The structures of their outer chain moieties comprise Galß1     

Lifetime list of publications by Hiroya Kawanabe

Hiroya Kawanabe produced more than 780 scientific papers, popular articles, governmental reports, chapters in books, edited or co-edited books, encyclopedia entries, and Japanese translations of books from 1952 until the compilation of this bibliography (Fall 1997). He was the sole or first author of 88% of these publications and shared authorship with 286 collaborators. Eighty-nine percent of his publications were in Japanese, others were in English, German, Chinese, Korean and Italian. His publications were devoted mainly to four topics: (1) ecology of freshwater fishes, especially the territorial behavior of ayu, Plecoglossus altivelis, and interspecific relationships and food segregation among fishes from 1956; (2) fish community ecology in Lake Tanganyika since 1979; (3) political articles on ecological research after 1991; and (4) museum activities after 1996. He also produced a number of newspaper articles (over 25% of total publications) addressing topics not only of science but literature, culture and philosophy. Kawanabe's articles in newspapers were produced mainly when he was the Director of the Center for Ecological Research of Kyoto University, and most of these articles concentrated on promoting the development of ecological research in Japan. The publications are arranged chronologically by year. The decision to add Japanese titles was based on the fact that their English translations, which follow in parentheses, are often too loose and imprecise. Unfortunately, our printers could not insert these titles in the Japanese characters so Roman transliterations had to be substituted.