Lectin uptake and incorporation into the calcitic spicule of sea urchin embryos

SummaryPrimary mesenchyme cells (PMCs) are skeletogenenic cells that produce a calcareous endoskeleton in developing sea urchin larvae. The PMCs fuse to form a cavity in which spicule matrix proteins and calcium are secreted forming the mineralized spicule. In this study, living sea urchin embryos were stained with fluorescently conjugated wheat germ agglutinin, a lectin that preferentially binds to PMCs, and the redistribution of this fluorescent tag was examined during sea urchin development. Initially, fluorescence was associated primarily with the surface of PMCs. Subsequently, the fluorescent label redistributed to intracellular vesicles in the PMCs. As the larval skeleton developed, intracellular granular staining diminished and fluorescence appeared in the spicules. Spicules that were cleaned to remove membranous material associated with the surface exhibited bright fluorescence, which indicated that fluorescently labelled lectin had been incorporated into the spicule matrix. The results provide evidence for a cellular pathway in which material is taken up at the cell surface, sequestered in intracellular vesicles and then incorporated into the developing spicule.

Observation about Architecture Skin Using New Material

This article researches the newly emerged architectural skin materials such as ETFE membranous material, paper, fabric, plastic, mental tapping board and mental weaving net, printing glass and colored glass, and various special materials. It analyses their characters and impression effects in architecture.

Histological and morphometric changes in untraumatised rabbit skeletal muscle treated with deep transverse friction

Deep transverse friction (DTF) is used in clinical practice and by its nature it may cause muscle injury. This study investigates the morphologic and morphometric changes in untraumatised rabbit skeletal muscle treated with DTF.Method: 16 New Zealand white rabbits were studied. The right vastus lateralis muscle was used as a control and the left vastus lateralis was treated with DTF. Muscle biopsies were taken 10 min, 24 h and 48 h after 1 treatment, 48 h after 2 treatments and 48 h and 6 days after 3 treatments. Treatments were 48 h apart. Biopsies were prepared for light microscopy and tissue morphometry.Results: After 1 DTF treatment, intracellular and extracellular oedema was noted. Contraction bands seen throughout the fibres suggested severe mechanical trauma to the muscle. 48 hours after 1, 2, and 3 treatments, the muscle appeared to be recovering with reduced oedema, and the contraction banding was limited to small focal areas throughout each fibre. Six days after the last treatment, the myofibers, although normal in diameter, showed small focal areas of super contraction and large internalised inclusion bodies composed of a pool of myofilaments or whorls of membranous material. Morphometry showed oedema to be maximal immediately after treatment.Conclusion: DTF causes a severe but reversible injury to untraumatized myofibers. Its possible mode of action in treatment of injured muscle requires further investigation.

Dictyostelium prespore-specific gene Dp87 encodes a sorus matrix protein

In this paper we report on the characteristics of the product of a prespore-specific gene (Dp87) of Dictyostelium discoideum. Polyclonal antibody was made against a bacterially synthesized Dp87-encoded protein fragment. Using this antibody, the product was characterized by immunochemical and immunocytological methods. It was shown that the Dp87-encoded protein is a prespore-specific protein with a molecular mass of 83 kDa, which first appears at the standing slug stage and persists in mature fruiting bodies. Western blot studies revealed the presence of an additional 81 kDa protein prior to the appearance of the 83 kDa protein from the tipped aggregate to the standing slug stage, thus indicating the former to be a precursor protein. Immunocytochemical and immunoelectron microscopic studies showed that the protein is bound to ER at the early stages of development when only the 81 kDa protein is present. At the later stages when the 83 kDa protein predominates, however, it becomes localized in prespore-specific vacuoles (PSVs) and is associated with the inner fibrous material of PSVs, but not with the peripheral membranous material. This is in contrast to spore coat proteins, which are localized in PSVs from the beginning of their appearance and associated with both structures of PSVs. In mature fruiting bodies, most Dp87 protein is localized to the interspore space (matrix) of the sori, with some left on the surface of the stalk tube. Disruptants of the Dp87 gene were also produced. Although they contained neither 81 kDa nor 83 kDa protein, they showed no phenotypic defects as compared to the parental strain.

Quantitation of protein 3 content of circulating erythrocytes at the single-cell level

The density and size of human erythrocytes has been roughly correlated with cell age, with the denser and smaller cells being older. Observations of this type have led to a hypothesis that the membranes of circulating erythrocytes are dynamic with respect to composition and that material is lost from the membrane during cell maturation and circulation. In this study, flow cytofluorimetry was used to investigate the distribution of the human erythrocyte anion transport protein (protein 3) in heterogeneous samples of circulating red cells. We verified that protein 3 can be specifically and quantitatively labeled in intact human erythrocytes with eosin-5-maleimide, a luminescent probe. Individual cells were accordingly analyzed for size by forward light scattering and for protein 3 content by quantitation of eosin fluorescence. Initial results indicated that the smallest erythrocytes had a protein 3 content equal to that of the largest circulating erythrocytes. This result was independently verified by light scatter-activated cell sorting; direct measurement of cell diameters by microscopy verified that the cell sizes of erythrocytes showing the 10% greatest and 10% smallest light-scattering signal were indeed distinct. Independent analysis of the size-sorted erythrocytes for protein 3 content was accomplished by gel electrophoresis of stroma from 150,000 large and small erythrocytes. Quantitative scanning densitometry of silver-stained gels of prepared stroma showed that protein 3 content of each set of fractionated cells was equal and did not vary as a function of cell size. Taken in combination with the reported correlation between increasing red blood cell age and decreasing cell size, these results indicate that any loss of membranous material during the cell aging process is not random.

Quantitation of protein 3 content of circulating erythrocytes at the single-cell level

The density and size of human erythrocytes has been roughly correlated with cell age, with the denser and smaller cells being older. Observations of this type have led to a hypothesis that the membranes of circulating erythrocytes are dynamic with respect to composition and that material is lost from the membrane during cell maturation and circulation. In this study, flow cytofluorimetry was used to investigate the distribution of the human erythrocyte anion transport protein (protein 3) in heterogeneous samples of circulating red cells. We verified that protein 3 can be specifically and quantitatively labeled in intact human erythrocytes with eosin-5-maleimide, a luminescent probe. Individual cells were accordingly analyzed for size by forward light scattering and for protein 3 content by quantitation of eosin fluorescence. Initial results indicated that the smallest erythrocytes had a protein 3 content equal to that of the largest circulating erythrocytes. This result was independently verified by light scatter-activated cell sorting; direct measurement of cell diameters by microscopy verified that the cell sizes of erythrocytes showing the 10% greatest and 10% smallest light-scattering signal were indeed distinct. Independent analysis of the size-sorted erythrocytes for protein 3 content was accomplished by gel electrophoresis of stroma from 150,000 large and small erythrocytes. Quantitative scanning densitometry of silver-stained gels of prepared stroma showed that protein 3 content of each set of fractionated cells was equal and did not vary as a function of cell size. Taken in combination with the reported correlation between increasing red blood cell age and decreasing cell size, these results indicate that any loss of membranous material during the cell aging process is not random.

A light and electron microscopic study of microsporogenesis in Azolla microphylla

SynopsisMicrosporogenesis in cultured material of Azolla microphylla was studied with the light and transmission electron microscopes. The first formed sporangium, a megasporangium, aborts and several microsporangia develop below. Initially, a single sporogenous cell is present, surrounded by a single layered tapetum and the microsporangial wall. Subsequently, several sporogenous cells are connected by plasmodesmata. The microspore mother cells are less densely cytoplasmic than the tapetal cells. Callose-like material is deposited around the microspore mother cells, but disappears before meiosis. The tetrads of microspores contain well defined organelles but less dense cytoplasm than the surrounding periplasmodium. Electron dense material deposited on the plasma membrane of the microspores eventually forms the endospore. The unornamented exospore develops by continued deposition of electron dense material. Degeneration of the periplasmodium gives rise to membranous material which appears to form a template for the massulae.