Membranes of animal cells. II. The metabolism and turnover of the surface membrane

Turnover studies of the surface membrane and of cell particulate matter of L cells in tissue culture in logarithmic and plateau phase of growth have been made. The rate of incorporation of isotope into these fractions and the rate of fall of specific activities of labeled L-cell fractions have been observed. The following interpretation of the data appears most likely although other interpretations are possible. Growing and nongrowing cells synthesize approximately similar amounts of surface membrane and particulate material. In the growing cell the material is incorporated with net increases in substance. There is relatively little turnover. In the nongrowing cell newly synthesized material is incorporated, but a corresponding amount of material is eliminated so that there is turnover without net increase of substance. Our results suggest that there is no gross differential turnover between the protein, lipid, and carbohydrate of the surface membrane under the conditions of our experiments. Metabolic inhibitors or omission of amino acids in the culture medium lead to a decrease in synthesis of surface membrane and cell particulates and cause an equivalent decrease in the rate of degradation of surface membrane and of particulates; therefore the synthetic and degradative aspects of turnover appear to be coupled. As cultures of nongrowing cells in suspension or on a glass surface age, their synthetic and turnover capacities diminish. Our results suggest that the cell may exist in a nongrowing state with a level of synthesis similar to that of a growing cell. It can exist in this state with a high level of turnover.     

Centrosome structure and microtubule nucleation in animal cells.

Genetic studies in the budding yeast have led to the molecular characterization of gamma-tubulin associated proteins and to the identification of orthologues in animal cells. While the gamma-tubulin complex is more complex in animal cells than in budding yeast, its function is probably maintained throughout evolution. In this review we discuss some of the possible regulations of the nucleation activity in the light of the centrosome structure. A potential cross-talk between microtubule nucleation and centrosome duplication is suggested by some, still scarce, data.     

The structure of glycophorins of animal erythrocytes

Glycophorins are red cell membrane sialoglycoproteins, which contain multipleO-linked oligosacchride chains and carry most of the cell surface sialic acid. Due to this high content of sialic acid the glycophorins are strongly stained with periodic acid-Schiff (PAS) reagent after sodium sodicylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The term glycophorin was proposed initially for human red cell sialoglycoproteins [1,2] and now it is also used for sialoglycoproteins in animal red cell membranes. Furthermore, similar glycoproteins of non-erythrocyte origin have also been identified and given the same name [3], although the terms leukosialin and sialophorin were proposed for a major sialoglycoproteins of human leukocytes [4,5]. In this article the term glycophorin will be used only for sialoglycoproteins existing in the erythrocyte membrane.Glycophorins of human erythrocytes, carrying blood group MN, Ss and other determinants, have been thoroughly studied and their properties described in several review articles [3,6,7,8]. The aim of this article is to summarize studies carriedout on the structure of non-human glycophorins, although some data concerning human glycophorins are included for comparative purposes.Abbreviations SDS-PAGE sodium dodecylsulphate-polycrylamide gel electrophoresis - PAS periodic acid-Schiff - LIS lithium diiodosalicylate     

Coping with the inevitable: how cells repair a torn surface membrane

Disruption of the cell plasma membrane is a commonplace occurrence in many mechanically challenging, biological environments. 'Resealing' is the emergency response required for cell survival. Resealing is triggered by Ca2+ entering through the disruption; this causes vesicles present in cytoplasm underlying the disruption site to fuse rapidly with one another (homotypically) and also with the adjacent plasma membrane (heterotypically/exocytotically). The large vesicular products of homotypic fusion are added as a reparative 'patch' across the disruption, when its resealing requires membrane replacement. The simultaneous activation of the local cytoskeleton supports these membrane fusion events. Resealing is clearly a complex and dynamic cell adaptation, and, as we emphasize here, may be an evolutionarily primitive one that arose shortly after the ancestral eukaryote lost its protective cell wall.     

Membranes of animal cells. I. Methods of isolation of the surface membrane

Procedures have been devised for the isolation of the surface membranes of mouse fibroblasts (L cell) and a variety of other cells. The surface membranes are stabilized by various reagents in a hypotonic solution and are then removed intact or as large fragments with a Dounce homogenizer. The membranes are purified by differential centrifugation on solutions of sucrose or glycerol or on a column of fine glass beads. A trilaminar pattern can be seen in thin sections of the membrane in the electron microscope. Sufficient material can be conveniently obtained for chemical analyses.     

The scaling and structure of aquatic animal wakes

Animal generated water movements are visualized and quantifiedusing two-dimensional particle image velocimetry (PIV). Theresulting vector flow fields allow for the study of the distributionof velocity, vorticity and vortices. Structural and temporalaspects of animal-induced flows covering a range of Reynolds(Re) numbers between less than 1 to more than 10⁴ are presented. Maps of flow induced by continuous foraging and intermittentescape responses of tethered nauplius and copepodid stages ofthe marine copepod Temora longicornis offer insight in viscosity-dominatedflow regimes. Fast escape responses of the equally sized largestnauplius stage and the smallest copepodid stage are compared.The nauplius moves by generating a viscous flow pattern withhigh velocities and vorticity; the copepodid moves by usinginertial effects to produce a vortex ring with a rearward jetthrough the center. Larvae and small adult fish (zebra danio) use a burst-and-coast-swimmingmode at Re numbers up to 6,000, shedding a vortex ring withthe associated jet at the tail during the burst phase. Flowpatterns during the coasting phase differ between the smalllarvae and larger adults due to the changes in importance ofviscosity. A 12 cm long mullet swimming in a continuous mode generatesa chain of vortex rings with a backward undulating jet throughthe centers of the rings at Re numbers of 4 x 10⁴ in inertia-dominatedregimes. Our empirical results provide realistic insight in the scaleeffects determining the morphology of the interactions betweenanimals and water.     

The stem cells of animal tissues]

A brief historical review of concepts on stem cells is given with a stress made on the role of Russian scientists A. A. Maksimov and A. A. Zavarzin who have formulated key problems in biology of stem cells and cell renewal. Principal stages in current studies of stem cells are considered. An outline is given of traditional and recent trends in the investigations: comparative studies of tissue systems with stem cells; molecular genetic mechanisms of the self-maintainance and the differentiation of stem cells; stem cells as a subject of gene engineering.     

Membranes of animal cells. V. Biosynthesis of the surface membrane during the cell cycle

KB cells grown in suspension culture were synchronized by using a double thymidine block. At various times throughout the life cycle aliquots of cells were pulsed with ¹⁴C-L-leucine, ¹⁴C-D-glucosamine and ¹⁴C-choline for one hour periods. Surface membranes, cell particulates and soluble proteins were isolated and their ¹⁴C specific activities were determined. It was found that there was a marked increase in the rate of incorporation into surface membrane just after division. The pattern of incorporation was the same for all three isotopic precursors. The rate of incorporation of isotopic precursors into soluble proteins was constant throughout the cycle. Some increase in rate of incorporation of isotope into the particulate fraction was observed during division.     

Effects of the surface wettability and zeta potential of bioceramics on the adhesiveness of anchorage-dependent animal cells

Using calcium phosphate ceramics that have high biocompatibility with the living body, the effects of the surface characteristics of the bioceramics on cell adhesiveness were investigated. In the case of carriers with contact angles from 35° to 60°, the cell adhesiveness increased according to the increase in the wettability. Measurement of the zeta potentials of HAP-TCP sinters showed that these bioceramics had negative potentials from −2 mV to −6 mV. Electrochemical analysis suggested that the initial cell anchoring ratio (R_ia) and adhesive strength (F_a,enz) were affected by the surface ionic condition of the ceramic material. To clarify the effects of the surface potential of the ceramics on cell adhesiveness, the ceramic surface was modified chemically by means of various silane coupling reagents. The surface potential was regulated from −20 mV to +24 mV. Using these ceramics, the affinity and adhesiveness of the cells to the ceramics were found to be dominantly regulated by the surface potential. A negative potential was effective in increasing the adhesiveness, even though living cells have negative charges.