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Biomedical subjects

D Volkmann

Publications and source records attributed to D Volkmann.

At least 37 records · Page 2Linked to original sources
PubMed1999

Characterization of the unconventional myosin VIII in plant cells and its localization at the post-cytokinetic cell wall.

Myosins are a large superfamily of motor proteins which, in association with actin, are involved in intra- cellular motile processes. In addition to the conventional myosins involved in muscle contractility, there is, in animal cells, a wide range of unconventional myosins implicated in membrane-associated processes, such as vesicle transport and membrane dynamics. In plant cells, however, very little is known about myosins. We have raised an antibody to the recombinant tail region of Arabidopsis thaliana myosin 1 (a class VIII myosin) and used it in immunofluorescence and EM studies on root cells from cress and maize. The plant myosin VIII is found to be concentrated at newly formed cross walls at the stage in which the phragmoplast cytoskeleton has depolymerized and the new cell plate is beginning to mature. These walls are rich in plasmodesmata and we show that they are the regions where the longitudinal actin cables appear to attach. Myosin VIII appears to be localized in these plasmodesmata and we suggest that this protein is involved in maturation of the cell plate and the re-establishment of cytoplasmic actin cables at sites of intercellular communication.

Arabidopsis↗
PubMed1999

Statoliths motions in gravity-perceiving plant cells: does actomyosin counteract gravity?

Statocytes from plant root caps are characterized by a polar arrangement of cell organelles and sedimented statoliths. Cortical microtubules and actin microfilaments contribute to development and maintenance of this polarity, whereas the lack of endoplasmic microtubules and prominent bundles of actin microfilaments probably facilitates sedimentation of statoliths. High-resolution video microscopy shows permanent motion of statoliths even when sedimented. After immunofluorescence microscopy using antibodies against actin and myosin II the most prominent labeling was observed at and around sedimented statoliths. Experiments under microgravity have demonstrated that the positioning of statoliths depends on the external gravitational force and on internal forces, probably exerted by the actomyosin complex, and that transformation of the gravistimulus evidently occurs in close vicinity to the statoliths. These results suggest that graviperception occurs dynamically within the cytoplasm via small-distance sedimentation rather than statically at the lowermost site of sedimentation. It is hypothesized that root cap cells are comparing randomized motions with oriented motions of statoliths and thereby perceiving gravity.

Actomyosin↗
PubMed1999

Impacts of aluminum on the cytoskeleton of the maize root apex. short-term effects on the distal part of the transition zone

Using monoclonal tubulin and actin antibodies, Al-mediated alterations to microtubules (MTs) and actin microfilaments (MFs) were shown to be most prominent in cells of the distal part of the transition zone (DTZ) of an Al-sensitive maize (Zea mays L.) cultivar. An early response to Al (1 h, 90 μM) was the depletion of MTs in cells of the DTZ, specifically in the outermost cortical cell file. However, no prominent changes to the MT cytoskeleton were found in elongating cells treated with Al for 1 h in spite of severe inhibition of root elongation. Al-induced early alterations to actin MFs were less dramatic and consisted of increased actin fluorescence of partially disintegrated MF arrays in cells of the DTZ. These tissue- and development-specific alterations to the cytoskeleton were preceded by and/or coincided with Al-induced depolarization of the plasma membrane and with callose formation, particularly in the outer cortex cells of the DTZ. Longer Al supplies (>6 h) led to progressive enhancements of lesions to the MT cytoskeleton in the epidermis and two to three outer cortex cell files. Our data show that the cytoskeleton in the cells of the DTZ is especially sensitive to Al, consistent with the recently proposed specific Al sensitivity of this unique, apical maize root zone.

Journal Article↗
PubMed1998

Profilin is associated with the plasma membrane in microspores and pollen.

In higher plants, a large number of isoforms for the actin monomer-binding protein profilin have been identified, whereas other organisms express only few profilins. Furthermore, plant profilin isoforms are expressed in a tissue-specific manner. These observations raise questions concerning functional and locational differences between isoforms of plant profilins. In this paper, we introduce three polyclonal antisera and one monoclonal antibody developed against purified pollen profilins from Zea mays and against recombinant maize profilin. Immunoblot analyses of native profilins and four recombinant maize pollen profilin isoforms show that three of the antibodies display a preference for certain isoforms. In situ immunofluorescence of pollen of Zea mays and two developmental stages of microspores of Betula pumila indicates that all antibodies label plasma membrane-associated domains. Thus, we show that at least some profilin isoforms are located at a distinct subcellular domain within developing microspores and, less distinctly, in mature pollen. This contrasts previously reported uniform distributions throughout the cytoplasm of mature pollen and pollen tubes. The results are discussed in light of the large number of profilins co-expressed in plants and with reference to accumulating evidence for functional differences between profilin isoforms.

Animals↗
PubMed1998

Gravisensitivity of cress roots.

The minimum dose (stimulus x time [gs]) eliciting a visible gravitropic response, has been determined using continuous and intermittent stimulation and two different accelerations at 1 g and 0.l g. The minimum dose of 20-30 gs estimated for microgravity roots and of 50-60 gs for roots grown on a 1 g-centrifuge indicated a higher sensitivity of microgravity roots. Applying intermittent stimuli to microgravity-grown roots, gravitropic responses were observed after two stimuli of 13.5 gs separated by a stimulus free interval of 118 s. The curvature of microgravity-grown roots to lateral stimulation by 0.1 g was remarkably smaller than by 1g in spite of the same doses which were applied to the seedlings. Microscopic investigations corresponding to stimulations in the range of the threshold values, demonstrated small displacement (< 2 micrometers) of statoliths in root statocytes. Accepting the statolith theory, one can conclude that stimulus transformation has to occur in the cytoplasm in close vicinity to the statoliths and that this transformation system was affected during seedling cultivation in microgravity.

Brassicaceae↗
PubMed1998

Autotropism, automorphogenesis, and gravity.

Segments of organs that have undergone gravitropic curvature later straighten during the course of gravitropism or after the g-vector becomes randomized on a clinostat. Little is known about the mechanism underlying these and perhaps related phenomena which have been described with various overlapping terms such as autotropism, autotropic straightening, automorphosis, automorphogenesis, automorphic curvature, and gravitropic straightening. The types of phenomena that historically have been named by the above terms are reviewed critically with respect to an interaction with gravitropism. We suggest that the term "autotropism" should not be applied to the phenomenon of organ straightening that occurs during the course of gravitropism, since this straightening is part of a complex series of local growth adjustments overall through time, and since this phenomenon is not itself a tropistic response to a directional exogenous stimulus. It is suggested that the term autotropism should be used only for the phenomenon of organ straightening that occurs after the g-vector is randomized on a clinostat or withdrawn in the microgravity conditions of spaceflight. Usage of the term automorphogenesis is most appropriate for describing curvatures or orientations that result from morphological relationships such as in nastic curvatures.

Cotyledon↗
PubMed1998

Autonomic straightening after gravitropic curvature of cress roots.

Few studies have documented the response of gravitropically curved organs to a withdrawal of a constant gravitational stimulus. The effects of stimulus withdrawal on gravitropic curvature were studied by following individual roots of cress (Lepidium sativum L.) through reorientation and clinostat rotation. Roots turned to the horizontal curved down 62 degrees and 88 degrees after 1 and 5 h, respectively. Subsequent rotation on a clinostat for 6 h resulted in root straightening through a loss of gravitropic curvature in older regions and through new growth becoming aligned closer to the prestimulus vertical. However, these roots did not return completely to the prestimulus vertical, indicating the retention of some gravitropic response. Clinostat rotation shifted the mean root angle -36 degrees closer to the prestimulus vertical, regardless of the duration of prior horizontal stimulation. Control roots (no horizontal stimulation) were slanted at various angles after clinostat rotation. These findings indicate that gravitropic curvature is not necessarily permanent, and that the root retains some commitment to its equilibrium orientation prior to gravitropic stimulation.

Brassicaceae↗
PubMed1997

Statocyte polarity and gravisensitivity in seedling roots grown in microgravity.

Space experiments have offered a unique opportunity to analyse the mechanism of gravisensing in plant roots. It has been shown that the strict structural polarity of statocytes observed on the ground is perturbed in microgravity: the amyloplasts move towards the proximal half of the cell and, at least in some cases, the nucleus becomes located further away from the (proximal) plasma membrane. It has thus been demonstrated that the amyloplasts do not move freely in the cytoplasm. Experiments using cytochalasin B (or D) have indicated that these organelles are attached to the actin network, probably by motor proteins. These findings have led to a new hypothesis on gravisensing the basis of which is that the tension in the actin filaments resulting from interaction with the statoliths would be transmitted to stretch-activated ion channels located in the plasma membrane (Sievers et al., 1991, In: Lloyd (ed) The cytoskeletal basis of plant growth and form, Academic Press, London New York, pp 169-182). Recently, it has been shown that the sensitivity of roots grown under 1 g conditions in orbit is less than that of roots grown in microgravity or under simulated weightlessness on clinostats. Since the location of the amyloplasts in microgravity is different from that in 1 g, the greater sensitivity observed could be due to different tensions in the actin network.

Brassicaceae↗
PubMed1997

Nuclear components with microtubule-organizing properties in multicellular eukaryotes: functional and evolutionary considerations.

The nucleus and the microtubular cytoskeleton of eukaryotic cells appear to be structurally and functionally interrelated. Together they constitute a "cell body". One of the most important components of this body is a primary microtubule-organizing center (MTOC-I) located on or near the nuclear surface and composed of material that, in addition to constitutive centrosomal material, also comprises some nuclear matrix components. The MTOC-I shares a continuity with the mitotic spindle and, in animal cells, with the centrosome also. Secondary microtubule-organizing centers (MTOC-IIs) are a special feature of walled plant cells and are found at the plasma membrane where they organize arrays of cortical MTs that are essential for ordered cell wall synthesis and hence for cellular morphogenesis. MTOC-IIs are held to be similar in origin to the MTOC-I, but their material has been translocated to the cell periphery, perhaps by MTs organized and radiating from the MTOC-I. Many intranuclear, matrix-related components have been identified to participate in MT organization during mitosis and cytokinesis; some of them also seem to be related to the condensation and decondensation of chromatin during the mitotic chromosome cycle.

Animals↗
PubMed1997

The 14-3-3 protein interacts directly with the C-terminal region of the plant plasma membrane H(+)-ATPase.

Accumulating evidence suggests that 14-3-3 proteins are involved in the regulation of plant plasma membrane H(+)-ATPase activity. However, it is not known whether the 14-3-3 protein interacts directly or indirectly with the H(+)-ATPase. In this study, detergent-solubilized plasma membrane H(+)-ATPase isolated from fusicoccin-treated maize shoots was copurified with the 14-3-3 protein (as determined by protein gel blotting), and the H(+)-ATPase was recovered in an activated state. In the absence of fusicoccin treatment, H(+)-ATPase and the 14-3-3 protein were well separated, and the H(+)-ATPase was recovered in a nonactivated form. Trypsin treatment removed the 10-kD C-terminal region from the H(+)-ATPase as well as the 14-3-3 protein. Using the yeast two-hybrid system, we could show a direct interaction between Arabidopsis 14-3-3 GF14-phi and the last 98 C-terminal amino acids of the Arabidopsis AHA2 plasma membrane H(+)-ATPase. We propose that the 14-3-3 protein is a natural ligand of the plasma membrane H(+)-ATPase, regulating proton pumping by displacing the C-terminal autoinhibitory domain of the H(+)-ATPase.

14-3-3 Proteins↗
PubMed1997

Immunofluorescence detection of F-actin on low melting point wax sections from plant tissues.

We developed a simple and reliable technique for immunofluorescence detection of F-actin on microtome sections of plant tissues. For the first time, large numbers of plant cells from various tissues that pass through their developmental stages could be consistently visualized on one section from plant organs. n-Maleimidobenzoic acid N-hydroxysuccinimide ester-pretreated and formalin-fixed segments of plant roots and shoots were embedded in low melting point ester wax at 37C and sectioned on a microtome. After dewaxing and rehydration, microfilaments were visualized by indirect immunofluorescence technique with a monoclonal anti-actin antibody. The technique has been successfully used for visualization of tissue- and development-specific F-actin arrays in cells of Zea mays and Lepidium sativum root tips and of maize stem nodes.

Actin Cytoskeleton↗
PubMed1997

Rearrangements of F-actin arrays in growing cells of intact maize root apex tissues: a major developmental switch occurs in the postmitotic transition region.

Immunofluorescence labeling, using a monoclonal antibody developed against actin, revealed the relative abundance and rearrangements of F-actin arrays which occur in cells of the maize root apex as they make the developmental transition from proliferative growth in the meristem to a non-proliferative state in more mature root parts, and during the concomitant process of tissue differentiation. Cells in both the root cap and the quiescent center are depleted of F-actin, whereas it is abundant in cells of the central cylinder but less so in the cortex. The cortical cytoplasm associated with the endwalls of both mitotic and postomitotic cells is characterized by a more intense reactivity to the actin antibody than the longitudinal side walls. A major change in F-actin arrangement occurs in the transitional growth region interpolated between the meristem and the zone of rapid cell elongation. The location and nature of these F-actin rearrangements within the root suggest that the F-actin system might be involved in generating a force associated with the developmental transition of cells from their slow near-isotropic mode of growth close to the base of the meristem, to rapid anisotropic growth which is characteristic of the zone of cell elongation. This attraction notion was strongly supported using specific inhibitors of F-actin and myosin.

Actins↗
PubMed1997

Impact of taxol-mediated stabilization of microtubules on nuclear morphology, ploidy levels and cell growth in maize roots.

Direct contact of the radiating perinuclear microtubules (MTs) with the nuclear envelope was visualized with an immunogold technique using specific monoclonal tubulin antibody. The possibility that these perinuclear MT arrays are involved in establishing and maintaining nuclear organization during the interphase of cycling cells in maize root meristems was tested using taxol, a MT-stabilizing agent. Taxol not only stabilized all MTs against the action of the MT-disrupters colchicine and oryzalin but also prevented these agents from their usual induction of nuclear enlargement and decondensation of nuclear chromatin. On the contrary, nuclear size decreased and the chromatin became more compact in mitotically cycling cells of the taxol-treated root apices. Moreover, taxol prevented the stimulation, by colchicine and oryzalin, of the onset of the S phase in cells of the quiescent centre and proximal root meristem. Exposure of maize roots to taxol strongly decreased final cell volumes, suggesting that the more condensed nuclear chromatin is less efficient in genome expression and that this accounts for the restriction of cellular growth. All these findings support the hypothesis that MT arrays, radiating from the nuclear surface, are an essential part of an integrated plant 'cell body' consisting of nucleus and the MT cytoskeleton, and that they regulate, perhaps via their impact on chromatin condensation and activity, progress through the plant cell cycle.

Cell Division↗
PubMed1996

Preservation of viable biological samples for experiments in space laboratories.

Standard viable preservation methods for biological samples using low temperatures have been investigated concerning their storage capabilities under higher temperature levels than usual. For a representative set of organism classes (plants, mammalian cells, arthropods and aquatic invertebrates), the minimum appropriate storage conditions have been identified by screening storage temperatures at -196 degrees, -80 degrees, -20 degrees, +4 degrees, +20 degrees/25 degrees C for periods from 2 days to 4 weeks. For storage below 0 degree C, as a typical cryopreservative, dimethylsulfoxide (DMSO) was used. For some samples, the addition of trehalose (as cryopreservative) and the use of a nitrogen atmosphere were investigated. After storage, the material was tested for vitality. The findings demonstrated that acceptable preservation can be achieved under higher storage temperatures than are typically applied. Small, dense cultured plant cells survive for 21 d when moderately cooled (+4 degrees to -20 degrees C); addition of trehalose enhances viability at -20 degrees C. For mammalian cells, the results show that human lymphocytes can be preserved for 3 d at 25 degrees C, 7 d at 4 degrees C and 28 d at -80 degrees C. Friend leukaemia virus transformed cells can be stored for 3 d at 25 degrees C, 14 d at 4 degrees C and 28 d at -80 degrees C. Hybridoma cells can be kept 7 d at 4 degrees C and 28 d at -20 degrees C or -80 degrees C. Model arthropod systems are well preserved for 2 weeks if maintained at lower temperatures that vary depending on the species and/or stage of development; e.g., 12 degrees C for Drosophila imagoes and 4-6 degrees C for Artemia nauplii. For aquatic invertebrates such as sea urchins, embryonic and larval stages can be preserved for several weeks at +6 degrees C, whereas sperm and eggs can best be stored at + 4 degrees C for up to 5 d at maximum. These results enhance the range of feasible space experiments with biological systems. Moreover, for typical terrestrial preservation methods, considerable modification potential is identified.

Animals↗
PubMed1996

Graviresponse of cress roots under varying gravitational forces.

During the spacelab mission IML-2 threshold values concerning gravity controlled growth processes have been estimated in order to test the reciprocity law (dose = stimulus x time = constant) for the first time under exact physiological conditions. Cress seedlings have been cultivated from dry seeds under conditions of microgravity and on a 1 x g-centrifuge in the ESA-BIORACK. With the help of NIZEMI--the slow rotating centrifuge microscope--these seedlings have been stimulated by different doses ranging from 12 to 60 x g x s. Two different values of acceleration--0.1 x g and 1 x g--have been used. Graviresponses of the roots have been documented by video recording for 60 min under conditions of microgravity. The response of roots to accelerations of 0.1 x g was remarkably less than to 1 x g in spite of the same doses being applied to the seedlings. Roots cultivated under conditions of microgravity showed a higher sensitivity than those grown on the 1 x g-centrifuge. Displacement of statoliths in gravity perceiving cells was mainly less than 1 micron under the different stimulation procedures. These results together with results from former space flights do not confirm the validity of the reciprocity law. They indicate that transformation of the gravistimulus has to occur in close vicinity to the statoliths, probably mediated by the ground cytoplasm and the cytoskeleton suspended therein.

Acceleration↗
PubMed1996

Complete disintegration of the microtubular cytoskeleton precedes its auxin-mediated reconstruction in postmitotic maize root cells.

The inhibitory action of 0.1 microM auxin (IAA) on maize root growth was closely associated with a rapid and complete disintegration of the microtubular (MT) cytoskeleton, as visualized by indirect immunofluorescence of tubulin, throughout the growth region. After 30 min of this treatment, only fluorescent spots were present in root cells, accumulating either around nuclei or along cell walls. Six h later, in addition to some background fluorescence, dense but partially oriented oblique or longitudinal arrays of cortical MTs (CMTs) were found in most growing cells of the root apex. After 24 h of treatment, maize roots had adapted to the auxin, as inferred from the slowly recovering elongation rate and from the reassembly of a dense and well-ordered MT cytoskeleton which showed only slight deviations from that of the control root cells. Taxol pretreatment (100 microM, 24 h) prevented not only the rapid auxin-mediated disintegration of the MT cytoskeleton but also a reorientation of the CMT arrays, from transversal to longitudinal. The only tissue to show MTs in their cells throughout the auxin treatment was the epidermis. Significant resistance of transverse CMT arrays in these cells towards auxin was confirmed using a higher auxin concentration (100 microM, 24 h). The latter auxin dose also revealed inter-tissue-specific responses to auxin: outer cortical cell files reoriented their CMTs from the transversal to longitudinal orientation, whereas inner cortical cell files lost their MTs. This high auxin-mediated response, associated with the swelling of root apices, was abolished with the pretreatment of maize root with taxol.

Antineoplastic Agents, Phytogenic↗
PubMed1996

Gravisensitivity of cress roots: investigations of threshold values under specific conditions of sensor physiology in microgravity.

The minimum dose (dose = stimulus x time), one of three threshold values related to gravity, was determined under microgravity conditions for cress roots. Seedlings were cultivated on a 1g centrifuge in orbit and under microgravity, respectively. After continuous stimulation on a threshold centrifuge, minimum doses of 20-30 gs for microgravity roots and 50-60 gs for roots grown on a 1g centrifuge were estimated, which indicated that microgravity roots have a higher sensitivity than 1g roots. These results do not confirm the threshold value of 12gs which was determined for cress roots using the slow rotating clinostat. Following application of intermittent stimuli to microgravity-grown roots, gravitropic responses were observed after two stimuli of 13.5 gs separated by a stimulus-free interval of 118s. Generally, this demonstrates that higher plants are able to 'sum up' stimuli which are below the threshold value. Microscopic investigations of the cellular structure corresponding to stimulations in the range of the threshold value demonstrated a small displacement of statoliths in root statocytes. No significant correlation was observed between gravitropic curvature and statolith displacement. If the statolith theory is accepted, it can be concluded that stimulus transformation must occur in the cytoplasm in the near vicinity of the statoliths and that this transformation system--probably involving cytoskeletal elements--must have been affected during microgravity seedling cultivation.

Centrifugation↗