Effect of starvation on glucose transport and membrane fluidity in rat intestinal epithelial cells

Studies on the surface area of microvilli (MV), fluidity of brush border membranes (BBM) and -glucose uptake were carried out in rat intestinal epithelial cells (IEC) during progressive starvation and under re-feed conditions. The surface area of MV, fluidity of BBM and -glucose transport through IEC membranes showed an increase during starvation when compared to well-fed controls. Re-feeding experiments restored the control values of all the three parameters within a short time. The results showed that the increase in -glucose transport through IEC membranes during starvation is due to increased surface area of MV and increased fluidity of BBM.     

The contractility of elongated microvilli in early sea urchin embryos

Summary Elongated microvilli attach the early sea urchin embryo to the fertilization envelope and support it in a concentric position within the perivitelline space. The contractility of the elongated microvilli was demonstrated in several ways. (1) During normal cleavage, these microvilli change their length to adapt to the change in shape and numbers of blastomeres. (2) When treated with calcium-free sea water, embryos become eccentrically located and the microvilli extend further than normal on one side; when returned to normal sea water, the embryos become centered again. (3) Several agents cause the fertilization envelope to become higher and thinner than normal and the elongated microvilli to extend correspondingly if treated within ten min after fertilization. In some cases, both elongated microvilli and fertilization envelope return to normal size when returned to normal sea water. (4) Fertilization in a papain solution causes the elongated microvilli and the fertilization envelope to contract to the surface of the embryo. (5) Refertilization after the papain-induced contraction can bring about the elongation of these microvilli and the elevation of the fertilization envelope a second time. It was also shown that elongated microvilli are extended immediately upon fertilization, at the same time as the short microvilli. The firm adherence of the tips of elongated microvilli to the fertilization envelope by means of extracellular matrix fibers is shown in a high voltage electron microscope stereoimage. This allows us to understand why it is that when the elongated microvilli extend or contract, the fertilization envelope also extends and contracts accordingly.     

Small Espin: A Third Actin-bundling Protein and Potential Forked Protein Ortholog in Brush Border Microvilli

An ~30-kD isoform of the actin-binding/ bundling protein espin has been discovered in the brush borders of absorptive epithelial cells in rat intestine and kidney. Small espin is identical in sequence to the COOH terminus of the larger (~110-kD) espin isoform identified in the actin bundles of Sertoli cell–spermatid junctional plaques (Bartles, J.R., A. Wierda, and L. Zheng. 1996. J. Cell Sci. 109:1229–1239), but it contains two unique peptides at its NH₂ terminus. Small espin was localized to the parallel actin bundles of brush border microvilli, resisted extraction with Triton X-100, and accumulated in the brush border during enterocyte differentiation/migration along the crypt–villus axis in adults. In transfected BHK fibroblasts, green fluorescent protein–small espin decorated F-actin–containing fibers and appeared to elicit their accumulation and/or bundling. Recombinant small espin bound to skeletal muscle and nonmuscle F-actin with high affinity (K_d = 150 and 50 nM) and cross-linked the filaments into bundles. Sedimentation, gel filtration, and circular dichroism analyses suggested that recombinant small espin was a monomer with an asymmetrical shape and a high percentage of α-helix. Deletion mutagenesis suggested that small espin contained two actin-binding sites in its COOH-terminal 116–amino acid peptide and that the NH₂-terminal half of its forked homology peptide was necessary for bundling activity.     

Changes in cell surface primary cilia and microvilli concurrent with measurements of fluid flow across the rabbit corneal endothelium ex vivo

Primary cilia and microvilli have been reported on the mammalian rabbit corneal endothelium but their relationship to cell function is undefined. Six corneas from healthy 2 kg female albino rabbits were glutaraldehyde-fixed post mortem (15:00 h) or twelve corneal stroma-endothelial preparations incubated at 37 degrees C under an applied hydrostatic pressure of 20 cm H2O for 4 h prior to fixation. The corneal endothelium was assessed by quantitative scanning electron microscopy. Cells fixed immediately post mortem were decorated with small stubby microvilli (average 21 +/- 13/100 micron 2), and only 25% of the cells were decorated with primary cilia having an average length of 2.44 +/- 1.56 microns. Following 4 h ex vivo incubation with a phosphate-buffered Ringer solution, conspicuous microvilli developed to an average density of 40 +/- 19/100 micron 2 and primary cilia were found on 12% of the cells, having on average length of 2.27 +/- 1.38 microns. Following 4 h incubation in a bicarbonate-buffered Ringer solution, small stubby microvilli developed to a density of 49 +/- 18/100 micron 2, and 40% of the cells showed primary cilia with an average length of 4.31 +/- 1.93 microns; the net trans-endothelial fluid flow in the latter set was 60% greater. These studies indicate that the primary cilia on corneal endothelial cells might be responsive to fluid flow, but that mild mechanical and/or chemical stress could also be the cause of the change since the elaboration of primary cilia can be accompanied by microvilli as well.     

Cytoskeleton Structure and Composition in Choanoflagellates

The structure and composition of the cytoskeleton has been studied in Monosiga ovata (Protozoa: Order Choanofiagellida Kent 1880) using a combination of methods in association with light and electron microscopy. Supplementary observations are included for Desmarella moniliformis. The basal body of the single anterior flagellum is subtended proximally and at right angles by a second, non-flagellar basal body. The edges of the two basal bodies are connected by a fibrillar bridge. A long, narrow, striated, fibrillar rootlet extends posteriorly from the lower edge of the non-flagellar basal body towards the Golgi apparatus. It is associated throughout most of its length with the surface of a flattened sac. Rootlet microtubules pass radially from a ring of electron dense material which encircles the distal end of the flagellar basal body. These microtubules extend outwards for about one-third of the length of the cell. Within each collar tentacle is a longitudinal bundle of microfilaments composed of actin as illustrated by rhodamine-phalloidin staining for fluorescence microscopy. The base of each microfilament bundle is associated with one or more rootlet microtubules by fine fibrillar bridges. The attachment between microtubules and tentacle microfilaments is further demonstrated by their coordinated displacement when the cytoskeleton becomes dislodged. The role of the cytoskeleton in maintaining the position of the collar tentacles during interphase and cell division is discussed.     

Induced immunity against the mosquito Anopheles stephensi: reactivity characteristics of immune sera

This study shows the progression of immune responses in mice during five sequential immunizations with Anopheles stephensi mosquito extracts, characterized by ELISA, Western blot and immunohistochemistry. When exposed repeatedly to mosquito bites, control mice developed antibodies which reached titres of 1:10(5), reacted weakly in Western blot analysis and were localized to the salivary glands. Mice immunized with mosquito head plus salivary glands, midgut, ovary, fat body, midgut microvilli (Mv) and midgut basolateral plasma membrane (Blm), showed increased titre with each successive boost. Epitopes were shared between sera or were specific to the immunizing or heterologous extract. Anti-Mv and Blm sera recognized proteins labelled by anti-midgut serum and gave specific reactions with the midgut and head. Cross-reactivity was confirmed immunohistochemically.     

The corneal epithelial surface in the eyes of vertebrates: environmental and evolutionary influences on structure and function

The smooth optical surface of the cornea is maintained by a tear film, which adheres to a variety of microprojections. These microprojections increase the cell surface area and are thought to improve the movement of oxygen, nutrients, and metabolic products across the outer cell membranes. However, little is known of these structural adaptations in vertebrates inhabiting different environments. This field emission scanning electron microscopic study examined the cell density and surface structure of corneal epithelial cells across 51 representative species of all vertebrate classes from a large range of habitats (aquatic, amphibious, terrestrial, and aerial). In particular, we wished to extend the range of vertebrates to include agnathans and some uniquely Australian species, such as the Australian lungfish (Neoceratodus forsteri), the Australian galah (Eolophus roseicapillus), the Australian koala (Phascolarctos cinereus), and the rat-tailed dunnart (Sminthopsis crassicaudata). Epithelial cell densities ranged from 28,860 +/- 9,214 cells mm(-2) in the flathead sole Hippoglossoides elassodon (a marine teleost) to 2,126 +/- 713 cells mm(-2) in the Australian koala (a terrestrial mammal), which may indicate a reduction in osmotic stress across the corneal surface. A similar reduction in cell density occurred from marine to estuarine to freshwater species. The structure and occurrence of microholes, microplicae, microridges, and microvilli are also described with respect to the demands placed on the cornea in different environments. All species that spend significant periods out of an aquatic environment possess microvilli and/or microplicae. These include all of our species of Mammalia, Aves, Reptilia, Amphibia, and even one species of Teleostei (Australian lungfish). Well-developed microridges occur only in teleosts in high osmolarity environments such as marine or estuarine habitats. Clear interspecific differences in corneal surface structure suggest a degree of adaptive plasticity, in addition to some phylogenetic trends.     

Ultrastructural features of the egg envelope of silver carp, <Emphasis Type="Italic">Hypophthalmichthys molitrix</Emphasis> (Osteichthyes,Cyprinidae)

Synopsis We investigated the ultrastructure of the egg membrane surface (unfertilized egg) of Hypophthalmichthys molitrix using scanning electron microscopy. The eggs of silver carp like most other teleosts are surrounded by a relatively thick egg membrane, and have a type III micropyle at the animal pole. The micropyle is almost circular in shape and the micropyle canal is located in its center. The micropyle region is not flat. Round or oval accessory pores are also observed in the canal. The surface of zona radiata is wavy and uneven with a uniform distribution of almost round pores with lips. Microvilli like structure are found in the pore opening region.     

Movement on the cell surface of the gliding bacterium,Mycoplasma mobile,is limited to its head-like structure

Mycoplasma mobile cells glide on solid surfaces such as glass with a fast and continuous motion in the direction of the membrane protrusion (head-like structure) at one cell pole. To examine its cell-surface movement, a latex bead was attached to a cell and behavior in gliding was monitored. The bead was carried without movement relative to the cell body, suggesting that the cell does not roll around the cell axis and the surface movement is limited to a small area. A small percentage of cells showed an elongated head-like structure in an old batch culture. The head-like structure moved forward, sometimes leaving the cell body in one position, resulting in a stretching of this head-like structure. These results indicate that the head-like structure drags the cell body, leading us to conclude that the force for gliding is generated at the head-like structure.     

Microvillus inclusion disease: a genetic defect affecting apical membrane protein traffic in intestinal epithelium

The striking similarities between microvillus inclusions (MIs) in enterocytes in microvillus inclusion disease (MID) and vacuolar apical compartment in tissue culture epithelial cells, led us to analyze endoscopic biopsies of duodenal mucosa of a patient after the samples were used for diagnostic procedures. Samples from another patient with an unrelated disease were used as controls. The MID enterocytes showed a decrease in the thickness of the apical F-actin layer, and normal microtubules. The immunofluorescence analysis of the distribution of five apical membrane markers (sucrase isomaltase, alkaline phosphatase, NHE-3 Na⁺/H⁺ exchanger, cGMP-dependent protein kinase, and cystic fibrosis trans-membrane conductance regulator), showed low levels of these proteins in their standard localization at the apical membrane as compared with normal duodenal epithelium processed in parallel. Instead, four of these markers were found in a diffuse distribution in the apical cytoplasm, below the terminal web (as indicated by co-localization with F-actin and cytokeratin 19), and in MIs as well. The basolateral protein Na⁺-K⁺ATPase, in contrast, was normally localized. These results support the hypothesis that MID may represent the first genetic defect affecting apical membrane traffic, possibly in a late step of apical exocytosis.