Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

scholarly journals The behavior of fibroblasts from the developing avian cornea. Morphology and movement in situ and in vitro.

1975 ◽  
Vol 67 (2) ◽  
pp. 400-418 ◽  
Author(s):  
J B Bard ◽  
E D Hay
Keyword(s):  
Three Dimensional ◽  
Cell Movement ◽  
Cell Process ◽  
General Mechanism ◽  
Dividing Cells ◽  
Collagenous Stroma ◽  
Collagen Lattices

The early chick cornea is composed of an acellular collagenous stroma lined with an anterior epithelium and a posterior endothelium. At stage 27-28 of development (5 1/2 days), this stroma swells so that the cornea is 75-120 mum thick. At the same time, fibroblasts that originate from the neural crest begin to invade this stroma. Using Nomarski light microscopy, we have compared the behavior of moving cells in isolated corneas with the migratory activities of the same cells in artificial collagen lattices and on glass. In situ, fibroblasts have cyclindrical bodies from which extend several thick pseudopodia and/or finer filopodia. Movement is accompanied by activity in these cytoplasmic processes. The flat ruffling lamelli-podia that characterize these cells on glass are not seen in situ, but the general mechanism of cell movement seems to be the same as that observed in vitro: either gross contraction or recoil of the cell body (now pear shaped) into the forward cell process, or more subtle "flowing" of cytoplasm into the forward cell process without immediate loss of the trailing cell process. We filmed collisions between cells in situ and in three-dimensional collagen lattices. These fibroblasts show, in their pair-wise collisions, the classical contact inhibition of movement (CIM) exhibited in vitro even though they lack ruffled borders. On glass these cells multi-layer, showing that, while CIM affects cell movement, fibroblasts can use one another as a substratum. Postmitotic cells show CIM in moving away from each other. Interestingly, dividing cells in situ do not exhibit surface blebbing, but do extend filopodia at telophase. The role of CIM in controlling cell movement in vivo and in vitro is stressed in the discussion.

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

scholarly journals Distribution of Potato spindle tuber viroid in Reproductive Organs of Petunia During Its Developmental Stages

2014 ◽  
Vol 104 (9) ◽  
pp. 964-969 ◽  
Author(s):  
Yosuke Matsushita ◽  
Shinya Tsuda
Keyword(s):  
Developmental Stages ◽  
Cell Movement ◽  
Seed Transmission ◽  
Potato Spindle Tuber Viroid ◽  
Reproductive Organs ◽  
Infected Male ◽  
Infected Female ◽  
Reproductive Tissues ◽  
Healthy Female

Embryo infection is important for efficient seed transmission of viroids. To identify the major pattern of seed transmission of viroids, we used in situ hybridization to histochemically analyze the distribution of Potato spindle tuber viroid (PSTVd) in each developmental stage of petunia (flowering to mature seed stages). In floral organs, PSTVd was present in the reproductive tissues of infected female × infected male and infected female × healthy male but not of healthy female × infected male before embryogenesis. After pollination, PSTVd was detected in the developed embryo and endosperm in all three crosses. These findings indicate that PSTVd is indirectly delivered to the embryo through ovule or pollen during the development of reproductive tissues before embryogenesis but not directly through maternal tissues as cell-to-cell movement during embryogenesis.

Spatial- and temporal-restricted pattern for amelogenin gene expression during mouse molar tooth organogenesis

Development ◽  
1988 ◽  
Vol 104 (1) ◽  
pp. 77-85 ◽  
Author(s):  
M.L. Snead ◽  
W. Luo ◽  
E.C. Lau ◽  
H.C. Slavkin
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Gene Transcription ◽  
Developmental Stages ◽  
Three Dimensional ◽  
Molar Tooth ◽  
Specific Expression ◽  
Amelogenin Gene ◽  
Stage Of Development

Position- and time-restricted amelogenin gene transcription was analysed in developing tooth organs using in situ hybridization with asymmetric complementary RNA probes produced from a cDNA specific to the mouse 26 × 10(3) Mr amelogenin. In situ analysis was performed on developmentally staged fetal and neonatal mouse mandibular first (M1) and maxillary first (M1) molar tooth organs using serial sections and three-dimensional reconstruction. Amelogenin mRNA was first detected in a cluster of ameloblasts along one cusp of the M1 molar at the newborn stage of development. In subsequent developmental stages, amelogenin transcripts were detected within foci of ameloblasts lining each of the five cusps comprising the molar crown form. The number of amelogenin transcripts appeared to be position-dependent, being more abundant on one cusp surface while reduced along the opposite surface. Amelogenin gene transcription was found to be bilaterally symmetric between the developing right and left M1 molars, and complementary between the M1 and M1 developing molars; indicating position-restricted gene expression resulting in organ stereoisomerism. The application of in situ hybridization to forming tooth organ geometry provides a novel strategy to define epithelial-mesenchymal signal(s) which are believed to be responsible for organ morphogenesis, as well as for temporal- and spatial-restricted tissue-specific expression of enamel extracellular matrix.

scholarly journals Kinesin-14 motors drive a right-handed helical motion of antiparallel microtubules around each other

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Aniruddha Mitra ◽  
Laura Meißner ◽  
Rojapriyadharshini Gandhimathi ◽  
Roman Renger ◽  
Felix Ruhnow ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Three Dimensional ◽  
Helical Motion ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Torque Generation ◽  
Free Microtubules ◽  
20 Nm

Abstract Within the mitotic spindle, kinesin motors cross-link and slide overlapping microtubules. Some of these motors exhibit off-axis power strokes, but their impact on motility and force generation in microtubule overlaps has not been investigated. Here, we develop and utilize a three-dimensional in vitro motility assay to explore kinesin-14, Ncd, driven sliding of cross-linked microtubules. We observe that free microtubules, sliding on suspended microtubules, not only rotate around their own axis but also move around the suspended microtubules with right-handed helical trajectories. Importantly, the associated torque is large enough to cause microtubule twisting and coiling. Further, our technique allows us to measure the in situ spatial extension of the motors between cross-linked microtubules to be about 20 nm. We argue that the capability of microtubule-crosslinking kinesins to cause helical motion of overlapping microtubules around each other allows for flexible filament organization, roadblock circumvention and torque generation in the mitotic spindle.

scholarly journals Topology of Chromosomes 18 and X in Human Blastomeres from 3- to 4-Day-old Embryos

2005 ◽  
Vol 53 (3) ◽  
pp. 273-276 ◽  
Author(s):  
Jan Diblík ◽  
Milan Macek ◽  
Maria-Cristina Magli ◽  
Roman Krejčí ◽  
Luca Gianaroli
Keyword(s):  
Three Dimensional ◽  
Dimensional Sphere ◽  
Chromosome X ◽  
Sphere Model ◽  
Chromosome 18 ◽  
Vitro Fertilization ◽  
Signal Distribution ◽  
Human In Vitro Fertilization

The positions of chromosomes 18 and X fluorescence in situ hybridization signals were analyzed in blastomeres generated from human in vitro fertilization 3- to 4-day-old embryos after preimplantation screening of aneuploidy of chromosomes 13, 16, 18, 21, 22, X, and Y. Fluorescent signal localization compared with a three-dimensional sphere model of random signal distribution revealed significant differences, providing evidence of peripheral localization of chromosome 18 in aneuploid ( p=0.0013) and aneuploid/euploid blastomeres ( p=0.0011). No differences were found in localization of chromosome 18 in euploid and in chromosome X in euploid and aneuploid blastomeres.

Fabrication of functional three-dimensional tissues by stacking cell sheets in vitro

2012 ◽  
Vol 7 (5) ◽  
pp. 850-858 ◽  
Author(s):  
Yuji Haraguchi ◽  
Tatsuya Shimizu ◽  
Tadashi Sasagawa ◽  
Hidekazu Sekine ◽  
Katsuhisa Sakaguchi ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Sheets

scholarly journals RE-EVALUATION OF THE STRETCH SENSITIVITY HYPOTHESIS OF CRUSTACEAN HEARTS: HYPOXIA, NOT LACK OF STRETCH, CAUSES REDUCTION IN HEART RATE OF ISOLATED HEARTS

1993 ◽  
Vol 176 (1) ◽  
pp. 223-232
Author(s):  
J. L. Wilkens
Keyword(s):  
Heart Rate ◽  
Isolated Heart ◽  
Carcinus Maenas ◽  
Three Dimensional ◽  
Decapod Crustacean ◽  
Diastolic Filling ◽  
Elastic Recoil ◽  
Isolated Hearts

Decapod crustacean hearts are suspended by a three-dimensional array of alary ligaments. These ligaments are stretched during systole; diastolic filling via the ostia occurs as the ventricle is stretched by ligamental elastic recoil. There is no direct venous return to the hearts in these animals. In the present study, an isolated heart preparation with intact ligaments, hereafter called in situ, was used to evaluate the effects of artificially induced stretch on heart rate. Strongly beating in situ neurogenic hearts of the crab Carcinus maenas responded to direct perfusion of the ventricle with oxygenated saline and the attendant augmentation of natural stretch with a small increase in heart rate (fh); however, fh was well maintained for up to 15 min after eliminating stretch by cutting the alary ligaments. In contrast to crabs, high rates of artificial perfusion usually depressed fh in crayfish hearts. Crab heart rate falls during hypoxia and this is readily reversed by even low rates of perfusion with oxygenated saline. It is concluded that the gradual decline in fh of totally isolated in vitro hearts arises from the deepening intraventricular hypoxia experienced by the cardiac ganglion.

scholarly journals Live 3D imaging and mapping of shear stresses within tissues using incompressible elastic beads

2021 ◽  
Author(s):  
Alexandre SOUCHAUD ◽  
Arthur BOUTILLON ◽  
Gaëlle CHARRON ◽  
Atef ASNACIOS ◽  
Camille NOÛS ◽  
...  
Keyword(s):  
Shear Stress ◽  
Covalent Binding ◽  
Three Dimensional ◽  
Contour Method ◽  
Shear Stresses ◽  
Liquid Droplets ◽  
Mechanical Constraints

To investigate the role of mechanical constraints in morphogenesis and development, we develop a pipeline of techniques based on incompressible elastic sensors. These techniques combine the advantages of incompressible liquid droplets, which have been used as precise in situ shear stress sensors, and of elastic compressible beads, which are easier to tune and to use. Droplets of a polydimethylsiloxane (PDMS) mix, made fluorescent through specific covalent binding to a rhodamin dye, are produced by a microfluidics device. The elastomer rigidity after polymerization is adjusted to the tissue rigidity. Its mechanical properties are carefully calibrated in situ, for a sensor embedded in a cell aggregate and submitted to uniaxial compression. The local shear stress tensor is retrieved from the sensor shape, accurately reconstructed through an active contour method. In vitro, within cell aggregates, and in vivo, in the prechordal plate of the Zebrafish embryo during gastrulation, our pipeline of techniques demonstrates its efficiency to directly measure the three dimensional shear stress repartition within a tissue, and its time evolution.

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