Surface glycoproteins of differentiating neuroblastoma cells analyzed by lectin binding and flow cytometry.

Cell surface glycoproteins of mitotic neuroblastoma cells and cells differentiated by prostaglandin cyclic adenosine monophosphate treatment were quantified by flow cytometric analysis and specific fluorescent lectins. No differences in fluorescent lectin binding were seen between suspensions of mitotically active and differentiated N2AB-1 cells following exposure to either fluorescein (FL)-labeled soy bean agglutinin (FL-SBA) specific for N acetyl galactosamine or FL-concanavalin A (FL-CON A) which binds to mannose residues. These lectins, however, were shown to bind specifically to these cells as revealed by competitive blocking studies with hapten sugars. When FL Ulex europaeus (FL-UEA) specific for fucose was reacted with control or differentiated cells, no binding was seen even with an increased dose of lectin before or after enzyme treatment. However, differentiated N2AB-1 cells, reacted with FL-wheat germ agglutinin (FL-WGA) specific for N acetyl glucosamine, bound more FL-WGA than that seen for control cultures. Furthermore, specific sites for FL-WGA were shown to be saturable and were lost upon pretreatment of cells with neuraminidase. Neuraminidase pretreatment revealed masked sites for FL-CON A and FL-SBA since binding was increased at least twofold for these lectins on mitotic and differentiated cells. These data indicate that single cell measurements of surface glycoproteins can be made on living neural cells and that differentiation induces an increase in cell surface N-acetyl glucosamine residues.

[1]  J. Leary,et al.  Tetanus toxin binding to neuroblastoma cells differentiated by antimitotic agents. , 1986, Brain research.

[2]  P. Marini,et al.  Muscle reinnervation—III. Motoneuron sprouting capacity, enhancement by exogenous gangliosides , 1983, Neuroscience.

[3]  J. Bee.  A cytochemical study of lectin receptors on isolated chick neural retina neurons in vitro. , 1982, Journal of cell science.

[4]  K. Pfenninger,et al.  Lectin labeling of sprouting neurons. II. Relative movement and appearance of glycoconjugates during plasmalemmal expansion , 1981, The Journal of cell biology.

[5]  K. Pfenninger,et al.  Lectin labeling of sprouting neurons. I. Regional distribution of surface glycoconjugates , 1981, The Journal of cell biology.

[6]  J. Wood,et al.  Lectin cytochemistry of carbohydrates on cell membranes of rat cerebellum , 1981, Journal of neurocytology.

[7]  M. Glick,et al.  An uncommon fucosyl linkage in surface membranes of human neuroblastoma cells. , 1980, Biochemical and biophysical research communications.

[8]  M. Potier,et al.  Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-alpha-D-N-acetylneuraminate) substrate. , 1979, Analytical biochemistry.

[9]  J. Gurd,et al.  Developmental alteration of rat brain synaptic membranes. Reaction of glycoproteins with plant lectins , 1979, Brain Research.

[10]  J F Leary,et al.  Laser flow cytometric light scatter and fluorescence pulse width and pulse rise-time sizing of mammalian cells. , 1979, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[11]  R. Akeson,et al.  Identification of a high molecular weight nervous system specific cell surface glycoprotein on murine neuroblastoma cells. , 1978, Experimental cell research.

[12]  R. Sidman,et al.  Cell reassociation behavior and lectin-induced agglutination of embryonic mouse cells from different brain regions. , 1978, Experimental cell research.

[13]  H. Hansson,et al.  Ultrastructural localization of cell membrane GM1 ganglioside by cholera toxin. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[14]  F. Charalampous,et al.  Interaction of concanavalin A with differentiated and undifferentiated murine neuroblastoma cells. , 1977, Archives of biochemistry and biophysics.

[15]  M. Glick,et al.  Glycopeptides from surface membranes of neuroblastoma cells. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[16]  F. Jacob,et al.  5-bromodeoxyuridine-induced differentiation of a neuroblastoma. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[17]  A. Raedler,et al.  Developmental modulation of neuronal cell surface determinants. , 1986, Bibliotheca anatomica.

[18]  P. Kelly.  Nervous System Glycoproteins , 1984 .

[19]  G. Brückner,et al.  Wheat germ agglutinin indicates the commencement of neuronal differentiation in the embryonic brain. A comparative carbohydrate histochemical study. , 1981, Acta histochemica.

[20]  Y. Kimhi.  Nerve Cells in Clonal Systems , 1981 .

[21]  S. Kornfeld,et al.  Structure of Glycoproteins and Their Oligosaccharide Units , 1980 .

[22]  S. Denis‐Donini,et al.  Molecular and lectin probe analyses of neuronal differentiation. , 1980, Current topics in developmental biology.

[23]  H. Mahler.  Glycoproteins of the Synapse , 1979 .

[24]  R. U. Margolis,et al.  Structure and Distribution of Glycoproteins and Glycosaminoglycans , 1979 .

[25]  I. Pastan,et al.  Role of cell surface carbohydrates and proteins in cell behavior: studies on the biochemical reversion of an N-acetylglucosamine-deficient fibroblast mutant. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[26]  R. Jeanloz,et al.  The Biological Role of Sialic Acid at the Surface of the Cell , 1976 .

[27]  N. Sharon,et al.  The biochemistry of plant lectins (phytohemagglutinins). , 1973, Annual review of biochemistry.