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Studies on phosphoglycerate mutase in facial processes from rats
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P,+UIIOCLOr';AL ANTIHOt)IES RECO(GNISING STAGE AND REGION SPECIFIC EPITOPES (I+~ EMBRYOr;IC t~UUSE PALATAL EPITHELIAL CELLS. ~.J. Uixon~ P.W.J. Ferquson and A. White. Dept. of Cell ~ Structural t~ioloqy, University of P;anchester, Enqland.
CELL SURFACE GLYCOCONJUGATES AS MARKERS FOR THE ENTODERMAL CELL LINE IN CLEAVAGE STAGE EGGS OF CREPIDULA FORNICATA (MOLLUSCA). M.R.Dohmen. University of Utrecht Department of Zoology, Padualaan 8, Utrecht, The Netherlands.
During maf~alian development bilateral palatal shelves, arise from the maxillary processes, grow i n i t i a l l y v e r t i c a l l y but rapidly elevate to a horizontal position. The rledial edge epithelia contact and fuse to forT1 a seam which subsequently deqenerates. Simultaneously, the nasal epithelia d i f f e r e n t i a t e into pseudostrat i f i e d , c i l i a t e d cells and the oral into s t r a t i f i e d , squamous, keratinising cells. Characterisation of specific e p i t h e l i a l cell surface antigens would enable more accurate assessment of palatal cell lineage and the mechanism of mesenchyme signalling of e p i t h e l i a l d i f f e r e n t i a t i o n . Palatal epithelium, was useO as an immunoqen to produce monoclonal antibodies using conventional technioues. Antibodies were produced which reacted against medial edge e p i t h e l i a l cells and the entire palatal epithelium. A t h i r d antibody bound to the basal and superficial cells of the nasal palatal epithelium, but only to superficial cells of the oral epithelium Supported by grants from Wellcome Trust.
Molluscan development is characterized by a constant cleavage pattern which allows the determination of cell lineage from the first cleavage of the egg up to the formation of the definitive organs. In Crepidula fornicata, the cell lineage has been studied by E.G.Conklin in 1897 (J.Morphol. 13: 1-226). At the 68 cell stage the mesodermal cell line separates from the entodermal one and now I I precursor ceils of the entoderm can be designated. At this stage the surface of the entodermal cells starts to be covered with a layer of glycoconjugate material which reacts intensely and specifically with the lectins Concanavalin A and Lens culinaris hemagglutinin B, thus demarcating the entoderm precursor cells sharply from the adjacent ectodermal and mesodermal ceils, This easily detectable marker can be used profitably in studies aimed at elucidating the mechanism of entodermal determination and early gene expression.
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POLY N-ACETYL LACTOSAMINE DERIVATIVES IN BRANCHING MORPHOGENESIS. S Fleming, Department of Pathology, University of Southampton, SOUTHAMPTON, England.
STUDIES ON PHOSPHOGLYCERATEMUTASE IN FACIAL PROCESSES FROM RATS. G.Granstr~m, H.M~r!]~, L.P.Nilsson and G.Zellin.Department of Hfstology, University of Gothenburg, S-400 33 Gothenburg, Sweden.
Giycoconjugates containing fucosylated derivatives of poly N-acetyl lactosamine have been shown to be important during early embryogenesls. I have studied the expression of some of these carbohydrates during branching morphogenesis of the respiratory and urinary systems in human fetuses by using immunocytochemical methods. Monoclonal antibodies have been used to detect three fucosylated derivatives of type II blood group chains, namely, LE x, LE ~ and blood group substance H. Cryptic antigenic sites and the internal chain structure have been studied by digestion with speciflc exoglycosidases and, respectively, endogalactosidase. The ampulla of the branches of the tracheal and ureteric buds expressed LE x and LE y on a linear carbohydrate chain. This was lost in the differentiated structures, collecting ducts and bronchi which expressed blood group substance H.
Phosphoglycerate mutase (PGM,EC 2.7.5.3) in mammalian tissues show the existence of three isoenzymes. This i n v e s t i g a t i o n was undertaken to study PGM isoenzymes in an experimental model f o r c r a n i o f a c i a l development. Free dissected f a c i a l processes were obtained from 9-15 day embryonic rats. Isoenzyme d i s t r i b u t i o n of PGM was recorded by IEF and by gel f i l t r a t i o n . The isoenzyme s e n s i t i v i t y to reagents of the sulfhydryl groups and to heat treatment was studied. There was a developmental change from the type BB PGM from the 9th embryonic day to isoenzyme MB and MM at the 15th day. Isoenzyme development preceeded morphological signs of muscle d i f f e r e n t i a t i o n . The type BB PGM was not affected by -SH group reagents, the type MB was i n h i b i t e d by 50% and MM was f u l l y i n h i b i t e d . Type MB showed a greater heat l a b i l i t y than type MM. The results support a homodimeric and heterodimeric structure f o r PGM and support the suggestion that PGM subunits B and M are determined by two d i f f e r e n t gene l o c i . 122S
343
P,+UIIOCLOr';AL ANTIHOt)IES RECO(GNISING STAGE AND REGION SPECIFIC EPITOPES (I+~ EMBRYOr;IC t~UUSE PALATAL EPITHELIAL CELLS. ~.J. Uixon~ P.W.J. Ferquson and A. White. Dept. of Cell ~ Structural t~ioloqy, University of P;anchester, Enqland.
CELL SURFACE GLYCOCONJUGATES AS MARKERS FOR THE ENTODERMAL CELL LINE IN CLEAVAGE STAGE EGGS OF CREPIDULA FORNICATA (MOLLUSCA). M.R.Dohmen. University of Utrecht Department of Zoology, Padualaan 8, Utrecht, The Netherlands.
During maf~alian development bilateral palatal shelves, arise from the maxillary processes, grow i n i t i a l l y v e r t i c a l l y but rapidly elevate to a horizontal position. The rledial edge epithelia contact and fuse to forT1 a seam which subsequently deqenerates. Simultaneously, the nasal epithelia d i f f e r e n t i a t e into pseudostrat i f i e d , c i l i a t e d cells and the oral into s t r a t i f i e d , squamous, keratinising cells. Characterisation of specific e p i t h e l i a l cell surface antigens would enable more accurate assessment of palatal cell lineage and the mechanism of mesenchyme signalling of e p i t h e l i a l d i f f e r e n t i a t i o n . Palatal epithelium, was useO as an immunoqen to produce monoclonal antibodies using conventional technioues. Antibodies were produced which reacted against medial edge e p i t h e l i a l cells and the entire palatal epithelium. A t h i r d antibody bound to the basal and superficial cells of the nasal palatal epithelium, but only to superficial cells of the oral epithelium Supported by grants from Wellcome Trust.
Molluscan development is characterized by a constant cleavage pattern which allows the determination of cell lineage from the first cleavage of the egg up to the formation of the definitive organs. In Crepidula fornicata, the cell lineage has been studied by E.G.Conklin in 1897 (J.Morphol. 13: 1-226). At the 68 cell stage the mesodermal cell line separates from the entodermal one and now I I precursor ceils of the entoderm can be designated. At this stage the surface of the entodermal cells starts to be covered with a layer of glycoconjugate material which reacts intensely and specifically with the lectins Concanavalin A and Lens culinaris hemagglutinin B, thus demarcating the entoderm precursor cells sharply from the adjacent ectodermal and mesodermal ceils, This easily detectable marker can be used profitably in studies aimed at elucidating the mechanism of entodermal determination and early gene expression.
344
345
POLY N-ACETYL LACTOSAMINE DERIVATIVES IN BRANCHING MORPHOGENESIS. S Fleming, Department of Pathology, University of Southampton, SOUTHAMPTON, England.
STUDIES ON PHOSPHOGLYCERATEMUTASE IN FACIAL PROCESSES FROM RATS. G.Granstr~m, H.M~r!]~, L.P.Nilsson and G.Zellin.Department of Hfstology, University of Gothenburg, S-400 33 Gothenburg, Sweden.
Giycoconjugates containing fucosylated derivatives of poly N-acetyl lactosamine have been shown to be important during early embryogenesls. I have studied the expression of some of these carbohydrates during branching morphogenesis of the respiratory and urinary systems in human fetuses by using immunocytochemical methods. Monoclonal antibodies have been used to detect three fucosylated derivatives of type II blood group chains, namely, LE x, LE ~ and blood group substance H. Cryptic antigenic sites and the internal chain structure have been studied by digestion with speciflc exoglycosidases and, respectively, endogalactosidase. The ampulla of the branches of the tracheal and ureteric buds expressed LE x and LE y on a linear carbohydrate chain. This was lost in the differentiated structures, collecting ducts and bronchi which expressed blood group substance H.
Phosphoglycerate mutase (PGM,EC 2.7.5.3) in mammalian tissues show the existence of three isoenzymes. This i n v e s t i g a t i o n was undertaken to study PGM isoenzymes in an experimental model f o r c r a n i o f a c i a l development. Free dissected f a c i a l processes were obtained from 9-15 day embryonic rats. Isoenzyme d i s t r i b u t i o n of PGM was recorded by IEF and by gel f i l t r a t i o n . The isoenzyme s e n s i t i v i t y to reagents of the sulfhydryl groups and to heat treatment was studied. There was a developmental change from the type BB PGM from the 9th embryonic day to isoenzyme MB and MM at the 15th day. Isoenzyme development preceeded morphological signs of muscle d i f f e r e n t i a t i o n . The type BB PGM was not affected by -SH group reagents, the type MB was i n h i b i t e d by 50% and MM was f u l l y i n h i b i t e d . Type MB showed a greater heat l a b i l i t y than type MM. The results support a homodimeric and heterodimeric structure f o r PGM and support the suggestion that PGM subunits B and M are determined by two d i f f e r e n t gene l o c i . 122S