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Biomedical subjects

M Wilcox

Publications and source records attributed to M Wilcox.

At least 73 records · Page 4Linked to original sources

The Drosophila PS2 antigen is an invertebrate integrin that, like the fibronectin receptor, becomes localized to muscle attachments.

We establish that the position-specific antigen 2 (PS2), a Drosophila cell surface glycoprotein complex, is an invertebrate member of the vertebrate fibronectin receptor (integrin) family. New monoclonal antibodies show that in Drosophila embryos and larvae PS2 alpha subunits have a size of ca. 140 kd. Analysis of cDNA and genomic clones revealed that the canonical PS2 alpha subunit contains 1394 amino acids and has extensive homology to the heavy and light chains of integrin alpha subunits. The distribution of the PS2 antigen is regulated at the level of PS2 alpha subunit mRNA. In early Drosophila development the protein is restricted to mesoderm and appears to be involved in muscle attachment. We suggest that PS2, like vertebrate fibronectin receptors, mediates changes in cell shape and cell-extracellular matrix adhesion by binding to a basement membrane protein.

Amino Acid Sequence↗

Drosophila position-specific antigens resemble the vertebrate fibronectin-receptor family.

The Drosophila position-specific (PS) antigens are a family of cell surface glycoprotein complexes thought to be involved in morphogenesis. Their overall structures and biochemical properties are similar to those of a group of vertebrate receptors, including those for fibronectin, fibrinogen and vitronectin, and also the leukocyte antigens Mac-1, LFA-1 and p150,95 and the VLA family of cell surface antigens. The N-terminal sequences of the alpha subunits of some of these molecules are homologous to the N-terminus of a PS antigen component. The Drosophila PS antigens thus appear to be homologous to these vertebrate receptors.

Animals↗

Distribution of Ultrabithorax proteins in Drosophila.

We have used a monoclonal antibody to examine the distribution of Ultrabithorax (Ubx) proteins in Drosophila embryos and imaginal discs by immunofluorescence. Ubx proteins are nuclear and show a spatially restricted distribution in the nervous system, epidermis and mesoderm. Labelling extends from the first thoracic segment (T1) to the eighth abdominal segment (A8) in the midline cells, from T2 to A8 in the ventral nervous system and epidermis and from A1 to A8 in the somatic mesoderm. In the nervous systems and epidermis the patterns of labelling exhibit a repeat unit, the Ubx metamere, that is out of phase with the segmental repeat unit. At least in the epidermis this repeat unit appears to extend between anterior-posterior compartment boundaries and consists of a posterior compartment together with the succeeding anterior compartment. The most prominently labelled metamere in the nervous system and epidermis is that comprising the posterior region of T3 and the anterior region of A1. Within each metamere the nuclei are heterogeneously labelled. Clear heterogeneity of labelling is also seen amongst the nuclei of the T3 imaginal discs.

Journal Article↗

Stimulated drug uptake in a photoreceptor cell.

In many cells, organelle translocation requires structural integrity of microtubules [2,6,11,14, 5]. We have found that extracellular application of the antimicrotubular drug colchicine, which was expected to enter cells simply by diffusion, was ineffective at blocking pigment granule migration (PGM) inside fly photoreceptors. However, illumination of these receptors in the presence of colchicine resulted in complete PGM block. From a combined study using microphotometry, microfluorometry and intracellular recordings, we inferred that light permeabilized the cell to the drug. This hypothesis was supported by the observation that illumination caused the photoreceptor to take up an inert dye. The results show that target cells optically selected from a homogeneous population can be induced to take up an agent from the extracellular fluid. Remarkably, this process of light-induced drug uptake does not affect cell viability.

Animals↗

Illumination induces dye incorporation in photoreceptor cells.

Illumination of fly photoreceptors in the presence of the fluorescent dye Lucifer yellow initiates incorporation of the dye, which stains each cell down to its synaptic terminal. Unilluminated cells do not become stained. Experiments on animals in vivo show that selected cells can be stained without loss of viability. "Induced endocytosis" provides a plausible mechanism underlying this phenomenon.

Animals↗

The Drosophila position-specific antigens are a family of cell surface glycoprotein complexes.

Position-specific (PS)1 and PS2 monoclonal antibodies bind non-uniformly to the mature wing imaginal disc of Drosophila with respect to the boundary separating the dorsal and ventral developmental compartments. PS1 antibodies preferentially recognize dorsal cells, PS2 antibodies ventral cells. Antibodies of the two classes extract distinct sets of glycoproteins from an imaginal disc lysate. PS3 antibodies bind to both dorsal and ventral disc cells and extract both PS1 and PS2 glycoprotein sets together with an additional component. We show that the PS antigens are related multimeric glycoprotein complexes on the cell surface. PS3 antibodies recognize a glycoprotein present in all complexes, while PS1 and PS2 antibodies recognize unique components of their own complexes. Spatial and temporal correlations suggest the molecules may have a function in development.

Animals↗

Protein products of the bithorax complex in Drosophila.

A sequence from the Ubx 5' exon in the bithorax complex of Drosophila melanogaster was expressed as a fusion protein in bacteria. This protein was used to raise rabbit antisera and monoclonal antibodies. These antibodies detect antigens that, on protein blots and by immunofluorescence on whole mounts of imaginal discs, show the predicted segmental distribution of Ubx products. These products are predominantly, if not totally, localized in the cell nucleus. In the embryonic nervous system nuclei are labeled from the second thoracic segment to the eighth abdominal segment. There is no labeling in homozygous Df bxd100 embryos.

Animals↗

Related cell-surface antigens expressed with positional specificity in Drosophila imaginal discs.

We have isolated three classes of monoclonal antibodies against Drosophila cell-surface antigens that are expressed with positional specificity in imaginal discs. Comparison of immunofluorescence patterns with the wing-disc fate map reveals that expression of the antigens is not directly related to the specific type of cuticular structure that a cell will make upon differentiation but depends on the position of the cell in the undifferentiated disc epithelium. On mature wing discs, each class of position-specific (PS) antibody binds nonuniformly with respect to the dorso-ventral compartment boundary, with PS1 antibodies binding primarily to dorsal cells and PS2 antibodies, to ventral cells. Antibodies of the different PS classes extract similar but nonidentical sets of large glycoproteins from cell lysates, and antibodies of the most general class, PS3, recognize the PS1 and PS2 antigens in addition to PS3-specific components. Thus, the distributions and molecular characteristics of the PS antigens suggest that the molecules are structurally and functionally related to one another.

Animals↗

Identification of tubulin isoforms in the plasmodium of Physarum polycephalum by in vitro microtubule assembly.

The tubulins of the plasmodium of Physarum polycephalum have been identified by in vitro microtubule assembly from partially purified extracts of asynchronous microplasmodia and late G2 macroplasmodia. The plasmodial tubulin group comprised of 2 alpha tubulins (app. m.w. 51000 daltons) and 2 beta tubulins (app. m.w. 58000 daltons and 55000 daltons) and appeared to be identical with a group of polypeptides which are synthesized periodically in late G2. Two of the plasmodial tubulin subunits (one alpha and one beta) were identical to the Physarum amoebal tubulin alpha and beta subunits as characterised by 2D gel positions.

Electrophoresis, Polyacrylamide Gel↗

A position-specific cell surface antigen in the drosophila wing imaginal disc.

The antibody produced by the hybrid cell line DK. 1A4 recognizes an antigen present initially on all the epithelial cells of the D. melanogaster wing imaginal disc. This antigen becomes progressively restricted to cells in the dorsal region of the disc during the final larval instar. The presence of the antigen does not correlate with the specific adult structures to which the cells will eventually contribute, but rather with the position of the cells in the disc. In late discs, the line bounding the region in which the antigen persists corresponds to the boundary between the dorsal and ventral compartments are revealed by a clonal analysis of the undifferentiated disc. Together, these data suggest that the antigen's disappearance may be specific to the cells of the ventral compartment of the wing disc.

Animals↗

Differentiation within the gonads of Drosophila revealed by immunofluorescence.

The antibody produced by the hybrid cell line DA.1B6 binds to the diploid epithelial cells of Drosophila. In this paper, we describe the immunofluorescence-binding pattern of the antibody to the gonads. A bright sheath of fluorescence extends from the seminal vesicle onto the most proximal part of the adult testis. The only other significant binding to the organ is to the apical cells of the germinal proliferation centre, which fluoresce brightly in testes from adults and from third instar larvae. In the adult ovary, there is strong binding to the cells of the follicular epithelium, although this binding is reduced in the latter stages of follicle development. Soon after the formation of a follicle, a pair of epithelial cells at each pole of the follicle can be seen to fluoresce much more brightly than the other cells. This early differentiation is reflected in the morphogenetic behaviour of these polar cells as the follicle develops. The anterior pair are among the 'border cells' which migrate between the nurse cells to the anterior pole of the developing oocyte; and, when the follicular epithelium around the oocyte becomes columnar, the posterior pair of cells do not elongate as much as the surrounding cells.

Animals↗