Transverse Microfibrils Research Articles

Microstructure Evolution of Polyacrylonitrile-Based Fibers during Thermal Pre-Oxidation

In this work, pre-oxidized polyacrylonitrile fibers are treated with ultrasonic etching and solution etching to produce ultra-thin sections. The evolution of the fibers’ microstructure in the pre-oxidation process is observed, and the transformation model of the microstructure of the pre-oxidized fibers is proposed. Scanning electron microscopy and high-resolution transmission electron microscopy were used to observe the microstructure changes of the fibers. Fourier transform infrared spectroscopy and X-ray diffraction were used to observe the chemical structure transformation and crystallization degree of the fibers in the pre-oxidation process. The results revealed that pre-oxidized fibers exhibited a smooth surface, while their interior consisted of fibrils. The longitudinal microfibrils were connected by the transverse microfibrils and amorphous regions. The fracture morphology of the fibers shifted from ductile to brittle, and the cross-section gradually became smoother. The linear molecular chain of PAN transformed into a ring structure as pre-oxidation progressed, subsequently leading to the cross-linking of this ring structure into an orderly trapezoidal configuration. The connection between the fibrils was enhanced, and the fiber structure became more compact and stable.

Dynamic Structural Change of Plant Epidermal Cell Walls under Strain.

The molecular foundations of epidermal cell wall mechanics are critical for understanding structure-function relationships of primary cell walls in plants and facilitating the design of bioinspired materials. To uncover the molecular mechanisms regulating the high extensibility and strength of the cell wall, the onion epidermal wall is stretched uniaxially to various strains and cell wall structures from mesoscale to atomic scale are characterized. Upon longitudinal stretching to high strain, epidermal walls contract in the transverse direction, resulting in a reduced area. Atomic force microscopy shows that cellulose microfibrils exhibit orientation-dependent rearrangements at high strains: longitudinal microfibrils are straightened out and become highly ordered, while transverse microfibrils curve and kink. Small-angle X-ray scattering detects a 7.4nm spacing aligned along the stretch direction at high strain, which is attributed to distances between individual cellulose microfibrils. Furthermore, wide-angle X-ray scattering reveals a widening of (004) lattice spacing and contraction of (200) lattice spacing in longitudinally aligned cellulose microfibrils at high strain, which implies longitudinal stretching of the cellulose crystal. These findings provide molecular insights into the ability of the wall to bear additional load after yielding: the aggregation of longitudinal microfibrils impedes sliding and enables further stretching of the cellulose to bear increased loads.

Three-dimensional regular arrangement of the annular ligament of the rat stapediovestibular joint

The stapes footplate articulates with the vestibular window through the annular ligament. This articulation is known as the stapediovestibular joint (SVJ). We investigated the ultrastructure of adult rat SVJ and report here on the characteristic ultrastructure of the corresponding annular ligament. Transmission electron microscopy showed that this annular ligament comprises thick ligament fibers consisting of a peripheral mantle of microfibrils and an electron-lucent central amorphous substance that is regularly arranged in a linear fashion, forming laminated structures parallel to the horizontal plane of the SVJ. Scanning electron microscopy revealed that transverse microfibrils cross the thick ligament fibers, showing a lattice-like structure. The annular ligament was vividly stained with elastica van Gieson’s stain and the Verhoeff’s iron hematoxylin method. Staining of the electron-lucent central amorphous substance of the thick ligament fibers by the tannate-metal salt method revealed an intense electron density. These results indicate that the annular ligament of the SVJ is mainly composed of mature elastic fibers.

Progress in understanding the role of microtubules in plant cells

Microtubules have long been known to play a key role in plant cell morphogenesis, but just how they fulfill this function is unclear. Transverse microtubules have been thought to constrain the movement of cellulose synthase complexes in order to generate transverse microfibrils that are essential for elongation growth. Surprisingly, some recent studies demonstrate that organized cortical microtubules are not essential for maintaining or re-establishing transversely oriented cellulose microfibrils in expanding cells. At the same time, however, there is strong evidence that microtubules are intimately associated with cellulose synthesis activity, especially during secondary wall deposition. These apparently conflicting results provide important clues as to what microtubules do at the interface between the cell and its wall. I hypothesize that cellulose microfibril length is an important parameter of wall mechanics and suggest ways in which microtubule organization may influence microfibril length. This concept is in line with current evidence that links cellulose synthesis levels and microfibril orientation. Furthermore, in light of new evidence showing that a wide variety of proteins bind to microtubules, I raise the broader question of whether a major function of plant microtubules is in modulating signaling pathways as plants respond to sensory inputs from the environment.

Wall architecture in the cellulose-deficient rsw1 mutant of Arabidopsis thaliana: microfibrils but not microtubules lose their transverse alignment before microfibrils become unrecognizable in the mitotic and elongation zones of roots.

The rsw1 mutant of Arabidopsis thaliana has a single amino acid substitution in a putative glycosyl transferase that causes a temperature-dependent reduction in cellulose production. We used recently described methods to examine root growth by surface marker particles, cell wall structure by field emission scanning electron microscopy and microtubule alignment by immunofluorescence after the mutant is transferred to its restrictive temperature. We find that raising the temperature quickly accelerates root elongation in both wild type and mutant, presumably as a result of general metabolic stimulation, but that in the mutant, the rate declines within 7-8 h and elongation almost ceases after 24 h. Radial swelling begins at about 6 h in the mutant and root diameter continues to increase until about 24 h. The normal transverse alignment of microfibrils is severely impaired in the mutant after 8 h, and chemical inhibition of cellulose synthesis by 2,6-dichlorobenzonitrile causes a similar loss of orientation. After 24 h, microfibrils are not clearly visible in the walls of cells that would have been in the mitotic and early-elongation zone of wild-type roots. Changes in older cells are less marked; loss of transverse microfibril orientation occurs without disruption to the transverse orientation of cortical microtubules. The wild type shows none of the changes except for acceleration of elongation, which in its case is sustained. We conclude that microfibril alignment requires the normal functioning of RSW1 and that, in view of the effects of dichlorobenzonitrile, there may be a more general linkage between the rate of cellulose production and its proper alignment.

F-actin involvement in apical cell morphogenesis of Sphacelaria rigidula (Phaeophyceae): mutual alignment between cortical actin filaments and cellulose microfibrils

The polarized apical cells of Sphacelaria rigidula display a well-organized cortical F-actin cytoskeleton. This consists of bundles of actin filaments (AFs), assuming definite patterns of organization in different regions of the cell cortex. At the tip region of the apical dome the AFs appear randomly oriented, showing a diffuse fluorescence. Immediately below, at the base of the apical hemisphere, the AFs form a ring-like band around the plasmalemma transverse to the polar cell axis. The rest of the cell cortex is traversed by AFs showing an axial or slightly inclined or helical orientation. Examination of the apical cells of S. rigidula in appropriate thin sections revealed that the wall has a multi-layered structure. In the tip region of the apical dome the cell wall bears randomly oriented cellulose microfibrils (MFs), while in the basal part of the apical dome it is reinforced by a layer of densely arranged transverse MFs. As the cell grows at the apex, the transverse MFs are continuously displaced towards the cell base. Below the transverse MF layer, an additional layer with axial or slightly oblique MFs starts being depositing internally, on the tubular part of the cell. Externally to them, the layer of transversely oriented MFs remains visible. The above observations were confirmed in apical cells of S. tribuloides. MF orientation in the innermost wall layer of the apical cells coincides with that of the cortical AFs observed by fluorescence. This mutual alignment between AFs and MFs in a cell that lacks cortical microtubules (MTs) suggests that the AFs are involved in the oriented deposition of MFs. Experimental disruption of AFs with cytochalasin B caused abnormal MF deposition, a fact strongly supporting the above hypothesis. The transverse MFs forming at the base of the apical dome define the diameter and consequently the cylindrical shape of the apical cells. It is suggested that in the brown algal cells examined the AFs play a morphogenetic role similar to that of cortical microtubules in higher plant cells.

Connecting gene and hormone action to form, pattern and organogenesis: biophysical transductions

Abstract Morphological effects of mutations and hormones in shoots and flowers are common. Hence the causal chain between genome and phenotype must contain biophysical steps which link soluble control compounds to the production of specific solid tissue configuration. Extension of cylindrical form results from the iterated directional expansion of cell and organ surface. This is thought to be based on transverse microfibril and microtubule (MT) alignment in cells. It is proposed that the key biophysical link is a dynamic ‘self-cinching’ process involving shear between adjacent cortical MTs in a ring. The MT array is maximally stable when the MTs are transverse. Gene products and hormones could act on form by influencing this minimal energy process. Production of pattern is the iterated emergence of new primordia in specific relation to pre-existing ones. It is proposed that the key biophysical step here is the minimal energy buckling (folding) of the tunica layer as it develops excess surface adjacent to older primordia. This physical response can also initiate regular undulations, primordia, in originally uniform tissue. Gene products and hormones could influence meristem size or tissueyielding properties and thereby govern both pattern and organ number. Organogenesis, connecting pattern to form, is proposed to involve the destabilization and restabilization of MT alignment controls within, and among, cells. All three processes are analysed in terms of minimal energy configurations of tissue and cell components.

Effects of gibberellin and colchicine on microfibril arrangement in epidermal cell walls of Vigna angularis Ohwi et Ohashi epicotyls

Gibberellic-acid (GA3) treatment of azukibean epicotyls resulted in alterations of the direction of newly deposited microfibrils, on the cell walls. Cells having transverse microfibrils on the inner surface of the wall were observed more frequently in GA3-treated epicotyls than in untreated or water-treated ones. This effect of GA3 was negated by simultaneously supplied colchicine. A crossed polylamellate structure was observed in the inner portion of the walls of GA3-treated cells, but not in the inner portion of the walls of colchicine-treated cells. The wall formed under the influence of colchicine consisted of microfibrils running in the same direction.

Cell expansion and microfibril deposition inClosterium ehrenbergii

Cells ofClosterium ehrenbergii expanded for about 4.5 hr after vegetative cell division and about 3 hr after sexual cell division. Short cells were produced by the sexual expansion. At the early stage of the vegetative and the sexual expansion, most microfibrils were deposited transversely to the cell axis on the inner surface of the new wall. Then, bundles of microfibrils, running in various directions, overlaid the transverse microfibrils already deposited, at the late stage of the both types of expansion. The pattern of microfibril deposition changed from the transverse to the bundled pattern about 4 hr after the vegetative and about 2 hr after the sexual cell division, respectively. The change of pattern of microfibril deposition seemed to be highly correlated with cessation of cell expansion.

Microfibrils in primary and secondary wall growth develop trellis configurations

In primary growth, protoplasmic pressure expands the cell wall as its cellulosic framework is being built up and strengthened. Numerous micrographs provide evidence that as each lamella of the wall is differentiated, randomly spaced bonds tend to develop between its adjacent microfibrils. Apparently the number of bonds so developed, and the number retained unbroken subsequently, are related to the rate of cell wall extension.Between unbroken bonds, tensions associated with growth cause adjacent transverse microfibrils to bend successively to each side of their original general orientation. Thus, within a lamella they form lenticular, trellis-like configurations. Large extensions cause large local variations in direction; some microfibrils remain adjacent within groups, but the groups may be widely separated. Such microfibril configurations are generally incompatible with the multinet theory. Apparently also, the very large extensions of the lamellae formed soon after cell division can cause breaking of bonds, wide dispersion, and even breaking of microfibrils.In secondary growth, similar bonding between microfibrils occurs within lamellae and also between lamellae of the cell wall. Again, there is associated development of trellis configurations, but in this case it is due to swelling within the wall during lignification. Resulting changes in microfibrillar directions between bonded positions are much less extreme than in primary walls, but they are highly significant physiologically.

Arrangement of cellulose microfibrils in walls of elongating parenchyma cells.

The arrangement of cellulose microfibrils in walls of elongating parenchyma cells of Avena coleoptiles, onion roots, and celery petioles was studied in polarizing and electron microscopes by examining whole cell walls and sections. Walls of these cells consist firstly of regions containing the primary pit fields and composed of microfibrils oriented predominantly transversely. The transverse microfibrils show a progressive disorientation from the inside to the outside of the wall which is consistent with the multinet model of wall growth. Between the pit-field regions and running the length of the cells are ribs composed of longitudinally oriented microfibrils. Two types of rib have been found at all stages of cell elongation. In some regions, the wall appears to consist entirely of longitudinal microfibrils so that the rib forms an integral part of the wall. At the edges of such ribs the microfibrils can be seen to change direction from longitudinal in the rib to transverse in the pit-field region. Often, however, the rib appears to consist of an extra separate layer of longitudinal microfibrils outside a continuous wall of transverse microfibrils. These ribs are quite distinct from secondary wall, which consists of longitudinal microfibrils deposited within the primary wall after elongation has ceased. It is evident that the arrangement of cellulose microfibrils in a primary wall can be complex and is probably an expression of specific cellular differentiation.

Changes in cell wall organization resulting from surface growth in parenchyma of oat coleoptiles

The primary walls present during the phase of extension growth in oat coleoptiles possess an almost transverse microfibril orientation on their inner surfaces but on the outer surface the microfibrils are considerably disoriented from this direction, which is consistent with the concept of multi-net mechanism of growth. Coleoptile segments grown at 2°C to depress cell wall formation show no difference in orientation on their inner and outer surfaces; this is also considered to be consistent with the multi-net mechanism. It is shown that the longitudinal ribs of microfibrils present at the cell corners, and hitherto referred to as secondary thickening, are on the outer surface of the cell wall and are considered to arise from a disorientation of microfibrils as a result of multi-net growth. As a result of this microfibril disorientation there is a tendency for the pit fields to be reduced in area. After surface growth has ceased a secondary wall is formed with a well-defined helical organization distinctly different from that of the primary wall. The implications of these results in terms of previous investigations are discussed.

Cell-wall studies in the Chlorophyceae. II. A preliminary study of the effect of continuous illumination on wall structure in Cladophora rupestris.

Cultures of Cladophora rupestris under continuous illumination, for periods up to six months, have shown that the wall structure is modified as compared with control cultures with more normal illumination. This has been demonstrated by means of X-ray analysis and by polarization optics, and supported by ancillary observations oh both cultures. Under continuous illumination transverse microfibrils are laid down much more abundantly than are the longitudinal microfibrils. This may form the first step towards an understanding of the genesis of the crossed fibrillar structure of plant cell walls.