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At least 1,135 records · Page 63Linked to original sources
PubMed2002

Neurogenesis in the mossy chiton, Mopalia muscosa (Gould) (Polyplacophora): evidence against molluscan metamerism.

Neurogenesis in the chiton Mopalia muscosa (Gould, 1846) was investigated by applying differential interference contrast microscopy, semithin serial sectioning combined with reconstruction techniques, as well as confocal laser scanning microscopy for the detection of fluorescence-conjugated antibodies against serotonin and FMRFamide. The ontogeny of serotonergic nervous structures starts with cells of the apical organ followed by those of the cerebral commissure, whereas the serotonergic prototroch innervation, pedal system, and the lateral cords develop later. In addition, there are eight symmetrically arranged serotonergic sensory cells in the dorsal pretrochal area of the larva. FMRFamide-positive neural elements include the cerebral commissure, specific "ampullary" sensory cells in the pretrochal region, as well as the larval lateral and pedal system. In the early juvenile the cerebral system no longer stains with either of the two antibodies and the pedal system lacks anti-FMRFamide immunoreactivity. Outgroup comparison with all other molluscan classes and related phyla suggests that the cord-like, nonganglionized cerebral system in the Polyplacophora is a reduced condition rather than a primitive molluscan condition. The immunosensitivity of the pedal commissures develops from posterior to anterior, suggesting independent serial repetition rather than annelid-like conditions and there is no trace of true segmentation during nervous system development. Polyplacophoran neurogenesis and all other available data on the subject contradict the idea of a segmented molluscan stem species.

Animals↗
PubMedDate not supplied

Interference reflection microscopic study of dorsal root growth cones on different substrates: assessment of growth cone-substrate contacts.

The contact between dorsal root growth cones and substrates of type IV collagen, fibronectin, poly-L-lysine, and laminin were studied using interference reflection microscopy (IRM) coupled with detaching growth cones from these substrates and observing substrate-associated membrane (SAM). IRM images of growth cones on collagen and fibronectin indicate substantial dark areas of close association between the substrate and basal region of the growth cone. The thin membranous veils and lamellapodia appear bright, indicating a greatly decreased association with the substrate. Filopodia can appear either entirely dark or light but usually are dark interspersed with brighter areas indicating varying degrees of contact. IRM images of growth cones on polylysine and laminin suggest a decreased association between the basal region of the growth cone and the substrate. The appearance of veils and lamellapodia was similar to that observed on collagen and fibronectin. Observations of SAM indicate an increased degree of contact between growth cones and substrates of collagen and fibronectin compared to laminin and polylysine. Collectively these data indicate that simple increases in adhesion are not adequate to explain substrate preference and enhancement of neurite elongation. Overall decreases in adhesion may accomplish the same phenomena, suggesting that it is perhaps more important to consider the regional distribution of adhesive contacts in relation to growth cone movement.

Animals↗
PubMed2001

Influence of cationic lipids on the stability and membrane properties of paclitaxel-containing liposomes.

Paclitaxel (taxol) is a poorly soluble anticancer agent that is in widespread clinical use. Liposomes provide a less toxic vehicle for solubilizing the drug and increasing the therapeutic index of paclitaxel in model tumor systems. The role of liposome membrane composition in the stability of paclitaxel-containing formulations is understood partially for neutral and anionic liposomes, but poorly for other compositions. We investigated the effect of dialkyl cationic lipids on the stability and physical properties of paclitaxel-containing liposomes, using circular dichroism (CD), fluorescence spectroscopy, and differential interference contrast microscopy (DIC). DOTAP (1,2-dioleoyl-3-trimethylammonium propane), a cationic lipid used frequently for gene delivery, was combined at various ratios with dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), or distearoylphosphatidylcholine (DSPC). In the absence of DOTAP, the stability of liposomes containing > or =3 mol% paclitaxel was observed to follow the following rank order: DPPC >DSPC > DMPC. Increasing concentrations of DOTAP increased the physical stability of all compositions, and maximal stabilization was achieved at 30-50 mol% DOTAP, depending on the paclitaxel concentration and the acyl chain length of the phosphatidylcholine. The relationship between stability and mole fraction of DOTAP was complex for some compositions. DOTAP exerted a major fluidizing effect on DMPC, DPPC, and DSPC membranes, and the addition of paclitaxel at 3-8 mol% did not increase fluidity further. Studies of membrane phase domain behavior using the probe Laurdan (6-dodecanoyl-2-dimethylaminonaphthalene) indicated that both paclitaxel and DOTAP were miscible with the phosphatidylcholine phase. The physical events leading to destabilization of formulations are hypothesized to arise from concentration-dependent paclitaxel self-association rather than immiscibility of the membrane lipids. Given the increased incorporation and stability of paclitaxel in DOTAP-containing membranes and the potential for enhanced interaction with cells, cationic liposomes may provide a therapeutic advantage over previously described liposome formulations.

Antineoplastic Agents, Phytogenic↗
PubMed1990

Oogenesis: chromatin and microtubule dynamics during meiotic prophase.

Changes in the organization of germinal vesicle chromatin in mouse oocytes have been analyzed by fluorescence microscopy with respect to progressive stages of follicular development and the disposition of oocyte cytoplasmic microtubules. Four discrete patterns of chromatin organization exist in germinal vesicle (GV)-stage oocytes isolated from the ovaries of 21-25-day-old gonadotropin-primed mice. Analysis of ovarian cryosections stained with the DNA-binding fluorochrome Hoechst 33258 indicates that sequential changes in GV chromatin occur during folliculogenesis that result in the formation of a continuous perinucleolar chromatin sheath at the time of antrum formation. Specific alterations in the cytoplasmic microtubule complex of GV-stage oocytes were observed that correlate with chromatin patterns. The extensive cytoplasmic microtubule complex seen in oocytes of preantral follicles initially localizes to perinuclear areas of the ooplasm. This is followed by a progressive reduction in cytoplasmic microtubules and the appearance of prominent microtubule-organizing centers at the nuclear periphery. Coordinated nuclear and microtubular alterations also occur under in vitro conditions prior to progression of meiosis to prometaphase-1. The results are discussed with respect to the ongoing differentiation of the oocyte nucleus and the microtubule cytoskeleton during folliculogenesis in preparation for the resumption of meiosis.

Animals↗
PubMed2005

Development of a genetic transformation system using new selectable markers for fission yeast Schizosaccharomyces pombe.

We describe the development of a new transformation system, using multiple auxotrophic marker genes, for the fission yeast Schizosaccharomyces pombe. We developed three new auxotrophic marker genes (arg12(+), tyr1(+) and ade7(+)) and generated a new host strain, YF043, by Cre-loxP-mediated gene disruption. YF043 possessed six mutated biosynthetic genes (leu1-32, ura4-M190T, arg12::loxP, tyr1::loxP, ade7::loxP and his2::loxP). The combination of this host strain and the new selectable markers can be used for gene disruption using the same preexisting transformation systems. In addition, Sz. pombe vectors were constructed, containing selectable marker genes that complement the auxotrophies of YF043. These new vectors are available for gene disruption and heterologous protein expression in strain YF043. The new Sz. pombe host strain will be a useful tool for molecular genetic studies of Sz. pombe where multiple recombinant modifications or multiple mutations are needed.

Blotting, Western↗
PubMed2007

The 160 N-terminal residues of calnexin define a novel region supporting viability in Schizosaccharomyces pombe.

Protein secretion is a complex process that can be modulated by folding factors in the endoplasmic reticulum (ER), such as calnexin, a highly-conserved molecular chaperone involved in quality control. In Schizosaccharomyces pombe, calnexin (Cnx1p) is essential for cell viability. The calnexin/Cnx1p determinants required for viability have been mapped within the last 123 residues of its C-terminus. To better understand the role(s) of calnexin/Cnx1p in secretion, we screened for cnx1 mutants 'super-secreting' cellulase. We identified ss14_cnx1, a mutant secreting 10-fold higher levels of the glycoprotein cellulase than the wild-type strain. While cellulase did not interact with ss14_Cnx1p, the ratio of secreted activity/quantity for this enzyme was not affected, suggesting that the quality control of folding in the ER was adequate in the mutant strain. Surprisingly, the ss14_Cnx1p mutant is composed of the 160 N-terminal amino acids of the mature molecule, thus this mutant defines a novel calnexin/Cnx1p region supporting Sz. pombe viability. Interestingly, like viable mutants spanning the last 52 aa of calnexin/Cnx1p, the 160 N-terminal residues encoded by ss14_cnx1 also forms a complex with the essential BiP chaperone. These results reveal the so far unidentified importance of the N-terminal region of calnexin/Cnx1p.

Amino Acid Motifs↗
PubMed1999

Rotokinesis, a novel phenomenon of cell locomotion-assisted cytokinesis in the ciliate Tetrahymena thermophila.

The mechanism responsible for final cell separation at the end of cytokinesis is currently unknown. Knockout strains of the ciliate, Tetrahymena thermophila lacking the kinesin-II homologous molecular motors, Kin1p and Kin2p are paralyzed due to their complete loss of cilia and undergo frequent cytokinesis failures. Observations of live dividing cells revealed that cleavage furrow ingression is normal in kinesin-II double knockout cells until the final stage of cell separation (Brown et al., 1999). During closer inspection of dividing cells using video differential interference contrast microscopy, we found that wild-type cells undergo an extremely complex motile behavior near the end of cytokinesis. This process, which we have named rotokinesis, appears to facilitate the physical separation of daughter cells. Here we present recent work on Tetrahymena rotokinesis, and review studies in other organisms which suggest that the use of cell locomotion in the completion of cytokinesis is a general phenomenon of motile cell types.

Animals↗
PubMed1999

Venation pattern formation in Arabidopsis thaliana vegetative leaves.

Branching net-like structures are a trait common to most multicellular organisms. However, our knowledge is still poor when it comes to the genetic operations at work in pattern formation of complex network structures such as the vasculature of plants and animals. In order to initiate a causal analysis of venation pattern formation in dicotyledonous plant leaves, we have first studied its developmental profile in vegetative leaves of a wild-type strain of the model organism Arabidopsis thaliana. As landmarks of the complexity of the venation pattern, we have defined three main developmental parameters, which have been quantitatively followed in time: the ratios of (a) the length and (b) the number of branchpoints of the vein network with the surface of the lamina, which decrease in parallel as the leaf grows, only small differences existing between successive leaves, and (c) the number of hydathodes per leaf, which increases both during leaf expansion and from juvenile to adult rosette leaves. We next searched for natural variations in the first vegetative leaves of 266 ecotypes, finding only 2 which showed a venation pattern unequivocally different from that of the rest, Ba-1 and Ei-5, the latter displaying an extremely simple pattern that we have called Hemivenata. This phenotype, which is inherited as a monogenic recessive trait, is visible both in leaves and in cotyledons and seems to arise from a perturbation in an early acting patterning mechanism. Finally, we have screened for mutants with abnormal venation pattern but normally shaped leaves, concluding that such a phenotype is rare, since only one recessive mutation was obtained, extrahydathodes, characterized by the presence of an increased number of hydathodes per leaf.

Arabidopsis↗
PubMed2002

3D-DIASemb: a computer-assisted system for reconstructing and motion analyzing in 4D every cell and nucleus in a developing embryo.

A computer-assisted three-dimensional (3D) system, 3D-DIASemb, has been developed that allows reconstruction and motion analysis of cells and nuclei in a developing embryo. In the system, 75 optical sections through a live embryo are collected in the z axis by using differential interference contrast microscopy. Optical sections for one reconstruction are collected in a 2.5-s period, and this process is repeated every 5 s. The outer perimeter and nuclear perimeter of each cell in the embryo are outlined in each optical section, converted into beta-spline models, and then used to construct 3D faceted images of the surface and nucleus of every cell in the developing embryo. Because all individual components of the embryo (i.e., each cell surface and each nuclear surface) are individually reconstructed, 3D-DIASemb allows isolation and analysis of (1) all or select nuclei in the absence of cell surfaces, (2) any single cell lineage, and (3) any single nuclear lineage through embryogenesis. Because all reconstructions represent mathematical models, 3D-DIASemb computes over 100 motility and dynamic morphology parameters for every cell, nucleus, or group of cells in the developing embryo at time intervals as short as 5 s. Finally, 3D-DIASemb reconstructs and motion analyzes cytoplasmic flow through the generation and analysis of "vector flow plots." To demonstrate the unique capabilities of this new technology, a Caenorhabditis elegans embryo is reconstructed and motion analyzed through the 28-cell stage. Although 3D-DIASemb was developed by using the C. elegans embryo as the experimental model, it can be applied to other embryonic systems. 3D-DIASemb therefore provides a new method for reconstructing and motion analyzing in 4D every cell and nucleus in a live, developing embryo, and should provide a powerful tool for assessing the effects of drugs, environmental perturbations, and mutations on the cellular and nuclear dynamics accompanying embryogenesis.

Animals↗
PubMed1995

Localization of utrophin and aciculin at sites of cell-matrix and cell-cell adhesion in cultured cells.

Aciculin is a phosphoglucomutase-related cytoskeletal protein associated with dystrophin and/or utrophin in various tissues and cell types. Comparison of expression patterns for aciculin, dystrophin, and utrophin in cultured cells demonstrated that aciculin is coexpressed with utrophin, but not with dystrophin, in cultures of A7r5 smooth muscle cells and REF52 fibroblasts. Some other nonmuscle cells synthesized only trace levels of or no aciculin and utrophin. Aciculin was detected by immunoblotting in antiutrophin immunoprecipitates from A7r5 and REF52 cultured cells, indicating an association between these two proteins. The aciculin-utrophin complex in fibroblasts and smooth muscle cells was mostly resistant to Triton X-100 extraction and was detected predominantly in the Triton-insoluble fraction, enriched in actin and actin-associated proteins. By immunofluorescence both aciculin and utrophin were identified in a similar dot-like or streak-like pattern in A7r5 and REF52 cultured cells. Immunolocalization of utrophin in cultured fibroblasts and smooth muscle cells in combination with interference reflection microscopy demonstrated that utrophin staining was mostly codistributed, but not exclusively confined to the areas of focal adhesions, sites of closest cell attachment to the substrate. Double immunostaining of A7r5 and REF52 cells for aciculin and utrophin revealed a precise colocalization of both cytoskeletal proteins at focal adhesions and along microfilaments. Costaining of cultured fibroblasts and smooth muscle cells with antibodies against utrophin and major focal adhesion components, vinculin and talin, showed that utrophin is concentrated in focal adhesions both at initial stages of cell spreading and in well spread cells of nearly confluent monolayers. In MCF10 breast epithelial cells both utrophin and aciculin were localized at cell-cell adherens-type junctions. Our data show that utrophin is a cytoskeletal component of cell-matrix and cell-cell adhesions in various cultured cells. In certain cell types the aciculin-utrophin complexes may contribute to the linking actin filaments to the plasma membrane.

Actin Cytoskeleton↗
PubMed1997

Asymmetry in the distribution of free versus cytoskeletal myosin II in locomoting microcapillary endothelial cells.

Myosin II is required for normal amoeboid locomotion. In order to understand how myosin II elicits its effects on locomotive behavior, we have mapped myosin II-cytoskeleton interactions in locomoting endothelial cells. Bovine microcapillary endothelial cells were microinjected with fluorescently labeled myosin II, and the distribution of myosin II was imaged in the living cells by fluorescence microscopy. The same cells were then permeabilized with Triton X-100 and imaged again. The second set of images showed only myosin II that was associated with detergent-insoluble cytoskeleton. Dividing the image of retained myosin II by that of total myosin II produced a map of the extent to which myosin II was associated with the detergent-resistant cytoskeleton at any point in the cell. In cells migrating at the edge of a scrape wound, myosin II was preferentially retained in a region approximately 10 microm wide located just behind the cells' leading lamellipodia. Relatively little myosin II was retained in perinuclear cytoplasm. A vector representation of the distribution of total versus retained myosin II demonstrated that myosin II retention was sharply polarized with respect to locomotion, favoring the front of migrating cells. Myosin II-enriched cytoskeleton in this region may help polarize protrusive activity and/or move cytoplasmic bulk forward. Patches of myosin II retention were also observed in adherent tails of many cells, consistent with a role in pulling the rear of the cell forward.

Animals↗
PubMed1999

Cell surface receptors transmit sufficient force to bend collagen fibrils.

To better understand the dynamic interaction of cells with their surrounding extracellular matrix, chondrocytes and rat embryo fibroblasts were overlaid with individual collagen fibrils and observed with high-resolution video-enhanced differential interference contrast microscopy. Although the cells had a polygonal shape characteristic of nonmotile cells, they used processes usually associated with cell locomotion to acquire the collagen fibrils. Instead of being transported in a retrograde direction, fibrils on the dorsal cell surface were bent, and regions of the bent fibrils were shifted in diverse directions. A blocking antibody to the beta1 integrin subunit significantly inhibited collagen fibril acquisition and bending. Enhanced actin assembly was only occasionally associated with fibrils undergoing rearrangement. Considering that the relatively stiff collagen fibrils require the application of force to be bent, this study shows that cells with a polygonal morphology (as opposed to a polarized, motile shape) are capable of exerting force through the beta1 integrins on the dorsal surface of the cell. Analysis of the bending patterns indicates that fibril buckling was induced by retrograde force combined with regions held stationary and/or the fibrils were bent by forces acting in opposing directions.

Actins↗
PubMed1997

Cholinergic stimulation of lacrimal acinar cells promotes redistribution of membrane-associated kinesin and the secretory protein, beta-hexosaminidase, and increases kinesin motor activity.

The role of the microtubule-based motor, kinesin, in membrane trafficking has been investigated in resting and stimulated acinar cells from rabbit lacrimal gland, a cholinergically controlled secretory tissue. Microtubule-dependent motors from extracts of control and carbachol-treated acini were isolated by microtubule-affinity purification and their activity was determined using a video-enhanced differential interference contrast microscopy assay for microtubule gliding. The observation that carbachol treatment resulted in a 2.2-fold stimulation of the frequency of GTP-dependent microtubule gliding in fractions isolated by microtubule-affinity purification and GTP release suggested that kinesin was a target of carbachol-induced stimulation. Resolution of membranes from resting cells by fractionation on a sorbitol density gradient followed by partitioning analysis in a dextran-polyethyleneglycol two-phase system revealed that membrane-associated kinesin codistributed with Golgi-derived membranes, a post-Golgi secretory compartment designated Hex1, membranes from a trans Golgi network-like compartment, endoplasmic reticulum and a group of putative lysosomal membranes containing cathepsin B. Comparable fractionation of carbachol-treated acini showed that stimulation caused redistributions of membrane-associated kinesin, the secretory enzyme beta-hexosaminidase, and galactosyltransferase that appeared to reflect both a reorganization within the Golgi complex and a return of material to the Golgi complex from the secretory pathway. Our findings that carbachol promotes activation of lacrimal acinar kinesin as well as major shifts in kinesin-membrane association within the secretory pathway suggests that kinesin plays a major role in secretory vesicle assembly, apical secretion, and/or secretory vesicle membrane recycling in the lacrimal gland.

Animals↗
PubMed2001

Ancylostoma caninum: the finger cell neurons mediate thermotactic behavior by infective larvae of the dog hookworm.

Bhopale, V. M., Kupprion, E. K., Ashton, F. T., Boston, R., and Schad, G. A. 2001. Ancylostoma caninum: The finger cell neurons mediate thermotactic behavior by infective larvae of the dog hookworm. Experimental Parasitology 97, 70-76. In the amphids (anteriorly positioned, paired sensilla) of the free-living nematode Caenorhabditis elegans, the so-called finger cells (AFD), a pair of neurons, each of which ends in a cluster of microvilli-like projections, are known to be the primary thermoreceptors. A similar neuron pair in the amphids of the parasitic nematode Haemonchus contortus is also known to be thermoreceptive. The hookworm of dogs, Ancylostoma caninum, has apparent structural homologs of finger cells in its amphids. The neuroanatomy of the amphids of A. caninum and H. contortus is strikingly similar, and the amphidial cell bodies in the lateral ganglia of the latter nematode have been identified and mapped. When the lateral ganglia of first-stage larvae (L1) of A. caninum are examined with differential interference contrast microscopy, positional homologs of the recognized amphidial cell bodies in the lateral ganglia of H. contortus L1 are readily identified in A. caninum. The amphidial neurons in A. caninum were consequently given the same names as those of their apparent homologs in H. contortus. It was hypothesized that the finger cell neurons (AFD) might mediate thermotaxis by the skin-penetrating infective larvae (L3) of A. caninum. Laser microbeam ablation experiments with A. caninum were conducted, using the H. contortus L1 neuronal map as a guide. A. caninum L1 were anesthetized and the paired AFD class neurons were ablated. The larvae were then cultured to L3 and assayed for thermotaxis on a thermal gradient. L3 with ablated AFD-class neuron pairs showed significantly reduced thermotaxis compared to control groups. The thermoreceptive function of the AFD-class neurons associates this neuron pair with the host-finding process of the A. caninum infective larva and shows functional homology with the neurons of class AFD in C. elegans and in H. contortus.

Ancylostoma↗
PubMed1996

Insulin and IGF-I binding in isolated trout cardiomyocytes.

Insulin and IGF-I receptor binding were characterized in cardiac muscle cells isolated from the brown trout, Salmo trutta fario. Cardiomyocyte suspensions obtained by perfusion of ventricles with collagenase showed a high degree of viability as judged by trypan blue exclusion, LDH leakage, and morphology. Specific insulin binding was 2.88 +/- 0.28%/10 mg cells after overnight incubation at 4 degrees. Scatchard analysis indicated the presence of high affinity insulin binding sites with an apparent dissociation constant (Kd) of 0.285 +/- 0.043 nM and a binding site density of 1. 61 +/- 0.19 x 10(8)/mg cells. Specificity of insulin binding was determined by displacing labeled insulin with increasing concentrations of IGF-I, and the Kd value obtained was 4.77 +/- 2.82 nM, 17-fold higher than Kd values for displacement of insulin tracer by nonlabeled insulin. The percentage of IGF-I specific binding (6.70 +/- 1.42%/10 mg cells), affinity (Kd = 0.163 +/- 0.023 nM), and binding site density (4.00 +/- 1.13 x 10(8)/mg cells) were higher than those of insulin. Displacement curves of labeled IGF-I with nonlabeled insulin (Kd = 33.6 +/- 9.9 nM), indicated a high specificity of the IGF-I binding site. High concentrations of cold insulin and IGF-I were able to decrease markedly the specific binding to their own receptor. Incubation with cold IGF-I also induced a diminution in insulin binding in agreement with the lower specificity of the insulin receptor. These data suggest that insulin and IGF-I are able to down-regulate their own receptor number in cardiac muscle cells. The present results demonstrate that the isolated cardiac myocyte preparation from brown trout is a useful model for studying insulin and IGF-I binding in fish heart tissue.

Animals↗
PubMed1997

Studies of Structure and Local Wetting Properties on Heterogeneous, Micropatterned Solid Surfaces by Microinterferometry.

The microstructure and the local wettability of functionalized patterns is reported. The patterns are formed by the photolithographic structuring of multilayers of hairy rod polymers on glass/MgF2/SiO2 substrates. By application of quantitative reflection interference contrast microscopy (RICM), the RICM image analysis was improved by accounting for (i) multiple reflections and the finite illumination aperture in stratified systems and (ii) geometrical effects in wedge-shaped systems and by (iii) optimizing the contrast enhancement through adjustment of the MgF2 coating. The surface free energy of hairy rod films were determined by analysing the contact angle of partially wetting liquid droplets as a function of the surface tension of the liquid. Periodic lattices of hydrophobic domains separated by hydrophilic domains were applied to check the theory of Joanny and de Gennes [J. Phys. Chem. 81, 552-562 (1984)] of contact angle hysteresis at periodic arrays of pinning centers. Measurements of the shape of the triple line and the contact angle hysteresis by RICM confirmed the theoretical predictions. Copyright 1997 Academic Press.

Journal Article↗
PubMed2001

Spreading Dynamics of Polydimethylsiloxane Drops: Crossover from Laplace to Van der Waals Spreading.

The spreading dynamics of small polydimethylsiloxane (PDMS) drops was studied on substrates with varying surface energies. For experimental parameters near the wetting transition, we observed small PDMS drops of different drop volumes as a function of time using interference video microscopy. While for large drops the contact angle θ decreases with the well-established power-law relation θ approximately t(-0.3) (Tanner's law), the effect of dispersive van der Waals (VW) interactions must be taken into account when interpreting the evolution of small drops. Two signatures of the VW forces are observed. For a positive Hamaker constant, the disjoining pressure acts as an additional driving force, leading to an acceleration of droplet spreading as soon as the drop height becomes comparable to the range of the VW interactions. In addition, a precursor film forms ahead of the contact line, leading to an apparent volume loss, particularly noticeable for very small drops. Contact line pinning may be a problem and we describe its effect on our experimental results. We present a theory that discusses the interplay of surface tension and VW forces in the case of a spreading drop. This model predicts a new spreading regime for very thin drops, in agreement with our experimental results. Copyright 2001 Academic Press.

Journal Article↗
PubMed1996

Microtubule minus ends can be labelled with a phage display antibody specific to alpha-tubulin.

To investigate the orientation of alpha- and beta-tubulin heterodimers within microtubules, we cloned a phage display antibody to alpha-tubulin. The N-terminal 100 residues of alpha-tubulin were bacterially expressed and used to select clones from a large repertoire of antibody-expressing phagemid particles. One clone reacted with the expressed alpha-tubulin N terminus and native tubulin dimer but not with the expressed beta-tubulin N terminus. Electron microscopy showed 30 nm gold beads coated with the antibody binding to one end of brain microtubules. The beads bound to the minus ends of axonemes but not to the brain tubulin extensions from their plus ends. In sliding motility assays with a plus end directed motor, beads were pushed ahead of the microtubules. Our results indicate that an N-terminal epitope of alpha-tubulin is exposed only at the minus ends of microtubules.

Animals↗