Glycoconjugate unique to migrating primordial germ cells differs with genera.

Previous cytochemical studies showing that rat primordial germ cells (PGCs) possess a unique surface glycoconjugate containing terminal alpha-N-acetylgalactosamine were extended in this study to determine whether a similar distinctive glycoconjugate coats the surface of PGCs in the mouse. The results showed that mouse PGCs fail to react with peroxidase-conjugated lectins specific for localizing glycoconjugate with terminal N-acetylgalactosamine. All available lectin conjugates with affinity for other terminal sugars or internal sugar linkages also failed to stain mouse PGCs except for the conjugates that bind to alpha-fucose. One fucose-specific lectin conjugate stained only PGCs in the early mouse embryo but stained additional sites in more mature embryos and lost reactivity with PGCs after gestational day 14. Another fucose-specific conjugate stained PGCs until day 15, but with less selectivity, and a third such conjugate bound to several sites, but not to PGCs. The results suggest that the developmental mechanisms mediating cellular interaction, migration, and differentiation may be similar in different genera, but the specific structure of the cell surface glycoconjugate involved in these mechanisms differs.

Lectin binding beneath the epithelium and in smooth muscle cells in the developing bronchial tree.

Components of the subepithelial stratum in developing rat lung reacted transiently with Maclura pomifera agglutinin (MPA) and Aleuria aurantia agglutinin (OFA) conjugated to horseradish peroxidase. These lectins possess selective affinity for and serve to localize glycoconjugates (GCs) with terminal Gal/GalNAc and Fuc, respectively. Staining was strongest with both lectins in the proximal bronchial tree and decreased peripherally to growing buds where it was absent. MPA staining of subepithelial structures decreased from the pseudoglandular through the canalicular period and disappeared by the terminal sac stage. Disappearance of this subepithelial reactivity coincided with appearance of apical MPA-positive glycoconjugate in the canalicular period. OFA stained selectively a layer of flattened cells and a thin extracellular stratum under the epithelium of proximal bronchi in the canalicular period. This lectin affinity extended farther peripherally in the pseudoglandular interval and diminished thereafter. The layer of OFA-positive cells underlying the epithelium was identified immunohistochemically as immature smooth muscle. These muscle cells gained contractile protein while losing surface lectin reactivity during fetal development. The high iron diamine method localized sulfated GC in basement membrane of proximal respiratory passages in the fetal lung. The results attest to the involvement of specific GCs in mediating epithelial-mesenchymal cell interaction during critical stages of bronchial morphogenesis.

Distribution of glycoconjugates in the optic vesicle and optic cup.

Lectin histochemical methods and immunohistochemical techniques have been utilized to investigate and partially characterize glycoconjugates in the developing eye. Peanut-lectin-binding sites associated with radial glial cells were found in the diencephalon. In the optic primordia, binding sites associated with radial glia were masked by terminal sialic acid, and only reacted with peanut lectin when pretreated with sialidase. This finding indicates that glycoconjugates associated with diencephalic radial glia contain terminal galactose-beta-(1----3)N-acetyl galactosamine, but glycoconjugates associated with radial glia in the optic primordia contain sialic acid----galactose-beta(1----3)N-acetyl galactosamine. The selective distribution of galactose, N-acetyl galactosamine and fucose associated with radial glial cells has also been demonstrated. We postulate that these distributions mediate the shaping of the developing eye.

Lectin histochemistry of the embryonic heart: fucose-specific lectin binding sites in developing rats and chicks.

Glycoconjugates, particularly their sugar side chains, play important roles in embryonic development. Changes in cell-surface-associated glycoconjugates are known to affect cell differentiation, cellular interactions, and other developmental phenomena during embryogenesis. The embryonic heart goes through a series of complicated morphologic events during development. Of particular interest is morphogenesis of the outflow tract. This region of the embryonic heart originates from more than one cell population and undergoes a complex process of septation during formation of the great vessels. Histochemical analysis with a series of fucose-specific lectins conjugated to horseradish peroxidase has revealed the presence of a fucosylated glycoconjugate in the outflow tract of the developing heart. The results reveal further that the expression of the fucosylated glycoconjugate is stage-dependent and thus probably genetically regulated. The timing and distribution of staining with the lectin OFA suggest that this fucosylated glycoconjugate may play a role in directing the migration of neural crest cells into the heart and subsequent formation of the conus septum.

Lectin histochemistry of the embryonic heart: expression of terminal and penultimate galactose residues in developing rats and chicks.

Rat embryos at days 10-18 of gestation and chicken embryos at days 3-6 of incubation were fixed and processed for lectin histochemistry. The distribution of binding sites for a lectin from the peanut Arachis hypogaea (PNA) conjugated to horseradish peroxidase (HRP) was determined on tissue sections both before and after enzymatic cleavage of sialic acid with neuraminidase (sialidase). Endocardial cushion tissue in the rat, but not in the chick, reacted with PNA-HRP prior to digestion with sialidase. Endocardium of both species (12 and 13 days in rat, 5 and 6 days in chick), particularly at the level of endocardial cushions, reacted strongly with the sialidase-PNA sequence; this staining decreased markedly after day 14 of gestation in the rat. PNA binding sites capped by sialic acid were most abundant in the developing rat heart during the critical period of endocardial cushion formation and decreased as development proceeded. The marked changes in the appearance and distribution of cardiac cell and tissue glycoconjugates during cardiogenesis support the concept that rapid changes occur in the structure of complex carbohydrates during embryonic and fetal development. The findings also suggest that such glycosylation-related events may be species specific.

Presence of a unique glycoconjugate on the surface of rat primordial germ cells during migration.

This investigation was undertaken to examine the chemical nature of components on the surface of primordial germ cells (PGCs) possibly related to their directed migration during development. To this end, lectins conjugated to horseradish peroxidase were used as specific histochemical probes to characterize the structure of PGC cell surface glycoconjugates and changes in their composition during and after their migration in the rat embryo. A lectin specific for terminal N-acetylgalactosamine (GalNAc) from Dolichos biflorus intensely stained the cell surface and a perinuclear region assumed to be Golgi zone of PGCs only during their migration. With one exception, no other site in the embryo stained with this lectin as migration proceeded. These observations suggest that the GalNAc-containing glycoconjugates on the surface of PGCs may be of functional importance in regulating the guidance and locomotion of these cells during the course of their extensive migration.