Biomedical subjects

G M Wessel

Publications and source records attributed to G M Wessel.

40 records · Page 3Linked to original sources

PubMed1986

Two embryonic, tissue-specific molecules identified by a double-label immunofluorescence technique for monoclonal antibodies.

We identify two tissue-specific molecules in the sea urchin embryo by an immunofluorescence technique capable of co-localizing monoclonal antibodies on the same tissue section. The technique uses monovalent Fab-fluorochrome conjugates as secondary reagents to avoid cross-talk of subsequent antibody probes. Using this technique, we show that two cell surface molecules are expressed by different cell populations in the embryo. The technique is generally applicable for antibodies regardless of species or subtype specificity, and uses commercially available reagents. The technique provides sufficient amplification and resolution for analytical work, yet is rapid enough for screening procedures. As a fluorescent counterstain, use of the dye 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS) in the protocol provides a distinct fluorescent background staining of the tissue without interference with the specific antibody staining.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

PubMed1985

Sequential expression of germ-layer specific molecules in the sea urchin embryo.

Described are two germ-layer specific molecules that appear coincident with the formation of two germ layer cell lineages in the sea urchin embryo. Meso1 is a molecule of 380 kDa that is first detected at the time of primary mesenchyme cell delamination from the wall of the blastula. Endo1 is a molecule of 320 kDa that appears on endoderm cells at the time of archenteron formation a few hours after Meso1 appears. Both antigens are identified by monoclonal antibodies. The appearance of these antigens is described by immunofluorescence microscopy, and quantitative data on their localization has been obtained by ultrastructural immunoelectron microscopy. The synthesis of the molecules has been followed by pulse-chase immunoprecipitation. Meso1 is first expressed in trans Golgi-like saccules, is concentrated in peripheral low electron-dense vesicles, and is found throughout the plasma membrane of the mesenchymal cells and their filopodial extensions. Newly translated Meso1 can first be immunoprecipitated upon differentiation of the mesoderm cell lineage, and pulse-chase studies suggest that the determinant is the result of a post-translational modification. [35S]Methionine pulses early in development followed by a chase to the mesenchyme blastula or prism stage show that at least a portion of the molecule is translated well in advance of the mesenchyme blastula stage. Endo1, in contrast, does not appear to be translated until the onset of gastrulation, just preceding the post-translational expression of the Endo1 determinant. Endo1 is localized to the apical and basolateral cell surfaces of the midgut and hindgut. No label is detected in foregut cells, demonstrating a heterogeneity of cell populations within the endoderm cell lineage corresponding to a difference in morphology. In addition, Endo1 is shown to be the result of new transcription by the embryonic genome. Even though the function of neither molecule is known, together they show the spatial and temporal precision of differentiation that accompanies the formation of germ layers.

Animals↗

PubMed1984

Ontogeny of the basal lamina in the sea urchin embryo.

The patterns of expression for several extracellular matrix components during development of the sea urchin embryo are described. An immunofluorescence assay was employed on paraffin-sectioned material using (i) polyclonal antibodies against known vertebrate extracellular matrix components: laminin, fibronectin, heparan sulfate proteoglycan, collagen types I, III, and IV; and (ii) monoclonal antibodies generated against sea urchin embryonic components. Most extracellular matrix components studied were found localized within the unfertilized egg in granules (0.5-2.0 micron) distinct from the cortical granules. Fertilization initiated trafficking of the extracellular matrix (ECM) components from within the egg granules to the basal lamina of the developing embryo. The various ECM components arrived within the developing basal lamina at different times, and not all components were unique to the basal lamina. Two ECM components were not found within the egg. These molecules appeared de novo at the mesenchyme blastula stage, and remained specific to the mesoderm through development. The reactivity of antibodies to vertebrate ECM antigens with components of the sea urchin embryo suggests the presence of immunologically similar ECM molecules between the phyla.

Animals↗

PubMed1981

Intercellular recognition: quantitation of initial binding events.

The hypothesis that intercellular adhesion can be subdivided into two separable phenomena--an initial recognition event and a subsequent stabilization--is supported by the use of a cell binding assay that provides a quantitative measure of intercellular binding strengths. Radioactive single cells are brought into contact with cell monolayers at 4 degrees C in sealed compartments. The compartments are inverted and a centrifugal force is then applied to dislodge the probe cells from the monolayers. By varying the speed of centrifugation, the force maintaining associations between embryonic chicken neural retina cells was determined to be on the order of 10(-5) dyne. Topographic specificities of single neural retina cells for retinal monolayers from various regions of the retina were detected with this assay and corresponded to those observed in more traditional assays at 37 degrees C. Also observed were two time- and temperature-dependent stabilization processes in which the force required for dislodgment increased. One of the stabilization processes was sensitive to dinitrophenol and was inactive at 4 degrees C; the second was still active in metabolically blocked cells. The metabolic-dependent process resulted in interactions at least 13 times as strong as the initial binding. The metabolic-independent process resulted in about a 2-fold increase in binding strength and had a temperature dependence similar to that of membrane diffusional phenomena.

Animals↗