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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 159 (1979), S. 81-87 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Study of the fine structure of the macronucleus in Euplotes eurystomus, a ciliate protozoon, during various stages of the cell division cycle has yielded new information about intranuclear helices. They are frequently observed at the periphery of chromatin bodies or next to the nuclear envelope, and they appear to be a constituent of nucleoli. The fibril that forms a helix is about 11-15 nm thick, and torus profiles of helices cut in cross section are about 35 nm in diameter. In substructure the helix is composed of a thin strand 3-5 nm thick which is coiled to form the 11-15 nm fibril; so the helix is a super-coiled structure. The intranuclear helices are present in the macronucleus throughout the cell cycle. They do not show obvious changes of relative abundance nor changes of relative localization in the nucleus, with one exception: they were never observed in the diffuse zone of replication bands. Evidence is presented indicating that nuclear helices migrate to the cytoplasm through nuclear pores. Although the chemical composition of the Euplotes intranuclear helices is unknown, information in the literature on similar helices in Amoeba indicates that they contain RNA and not DNA. The observations on Euplotes helices are consistent with a concept of “packaged” RNA for transport to the cytoplasm.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 128 (1969), S. 369-385 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The fine structure of the thyroid of the newt, Notophthalmus viridescens, was studied with the electron microscope. Specimens were injected I.P. with 30 μc of I131 and sample thryoids were examined at 12 hour intervals thereafter. The ultrastructure of the normal thyroid gland is described, and compared with that of the irradiated glands. The first visible ultrastructural change observed after injection of the radioiodin was a striking alteration of nuclear morphology. This effect was followed by an increase in the frequency of whorled lamellar structures, a decrease in the number of microvilli, and degeneration of the Golgi apparatus and endoplasmic reticulum. Further effects observed included an increase in the number of large cytoplasmic granules and a decrease in the number of smaller ones, the presence of autophagic vacuoles, and finally, an increase in the number of degenerated mitochondria.
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  • 3
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The fine structure of the dorsal bristle complex and pellicle of non-developing Euplotes eurystomus is described in detail by scanning and transmission electron microscopy. The bristle-pit unit is a highly differentiated complex of organelles. The bristle complex is composed of a pair of kinetosomes (basal bodies) joined by a connective. The anterior kinetosome bears the bristle cilium, which contains a polarized network of particles (“lasiosomes”). The posterior kinetosome bears a very short, knob-like “condylocilium,” and has an associated striated fiber. Accessory ribbons of microtubules are also associated with the kinetosome couplets. Parasomal sacs, a septum connecting the bristle cilium to the anterior wall of the pit, core granules of the kinetosomes, and large membranous ampules are described. The organization of the bristle complex bears many similarities to the somatic ciliature of other ciliates. The pellicle of Euplotes is composed of a continucus outer cell membrane subtended by membranous alveoli, which contain a “fibrous mat.” Two sheets of subpellicular microtubules (longitudinal and transverse) are located just beneath the alveoli. The “epiplasm” seen in some other ciliates is apparently absent in Euplotes. The texture of the cell surface is a pattern of folds or rugae composed of the outer cell membrane and the upper membrane of the alveolus. The pattern of rugae probably defines the “silverline-system” of light microscopy.
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  • 4
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Hypotrichs are among the most complex ciliates in terms of morphology and development. To study the fine structure of cortical morphogenesis associated with cell division in Euplotes eurystomus, three different methods of observation were employed: light microscopy of protargol-stained specimens, scanning electron microscopy of cells prepared by critical point drying, and transmission electron microscopy of sectioned material. Observations on the stages of morphogenesis give much new information about cortical development, particularly about proliferation and aggregation of kinetosomes (basal bodies), ciliary outgrowth, the topography of morphogenesis, cirrus resorption, and growth of the pellicle. During the formation of new cirrus the process of kinetosome proliferation is atypical, i.e., groups of prokinetosomes are seen oriented at random and, in some cases, prokinetosomes apparently are formed at a distance from nearby young kinetosomes. That the new cirri develop in surface grooves, the grooves elongate into “tracks,” and (in some cases) grooves are partitioned into separate tracks suggests that the grooves play a role in the orderly migration of the new cirri on the cell surface. Conspicuous morphogcnctic changes in the cell surface involve local growth of the pellicle. The process of pellicle growth apparently involves two basic steps: (a) growth of the outer cell membrane to form “bare regions,” and (b) formation of alveoli in the bare regions. Alveolar sheets are formed by fusion of alveolus precursor particles. Cirrus resorption is sequential over several stages of development, and old cirri are resorbed as the new cirri impinge on them. As the old cirri regress, both in situ resorption and retraction of axonemes into the cytoplasm occur.
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  • 5
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The differentiation and degeneration of the cement gland in Xenopus laevis is described. The gland is first observed histologically at stage 19 (neural tube stage) as a packed group of apical ectoderm cells heavily laden with oocyte pigment granules, lying ventral to the cranial neural fold. By tailbud stage 35/36, the gland cells have increased in height and are approximately ten times taller than nonglandular apical ectoderm cells. The nuclei divide the gland cells into an apical region that is eosinophilic and contains oocyte pigment granules, and a basal region that contains clear droplets. The cells are decreasing in height by stage 40 (early tadpole) and begin to lose their pigment granules. Between stages 45 and 48, the pigment is extruded and the clear basal droplets diminish in number. From stage 48 to 49 the cells become vacuolated and the histotypic characteristics of the functional gland are lost. The gland is not vascularized, nor do phagocytic cells appear in its vicinity during any stage of its development. It remains bordered at its base by subjacent basal ectoderm during its entire life cycle of 10 to 12 days at room temperature.
    Additional Material: 1 Tab.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 202 (1989), S. 29-51 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The developing skeleton of a pelobatid frog, Spea bombifrons, is described based on a series of 81 cleared-and-stained tadpoles and juveniles. Most limb and limb-girdle elements commence ossification premetamorphically, along with the parasphenoid, frontoparietals, exoccipitals, and vertebral column, during Gosner Stage 36. The chondrocranium undergoes dramatic restructuring at the beginning of metamorphosis (ca. Stage 40); and the nasal cartilages, prootics, premaxillae, nasals, maxillae, septomaxillae, and ischium appear. Near the end of metamorphosis (ca. Stage 44), the branchial arches are resorbed, the hyoid plate and quadrate form, and the angulosplenials, vomers, squamosals, dentaries, and pterygoids ossify. After metamorphosis (Stage 46), the laryngeal cartilages, sternum, omosternum, and plectral apparatus chondrify; and the carpals, tarsals, sphenethmoid, posteromedial hyoid processes, mentomeckelian bones, quadrates, columellae, and opercula ossify. The development of the fused sacrococcygeal articulation in S. bombifrons is described and compared to the more widespread bicondylar articulation. The presumed palatine bone and transverse processes of the coccyx are discussed, as are several seemingly paedomorphic skeletal features of Spea. Comparison with the sequence and timing of ossification in other anurans reveals that ossification of the skull and vertebral column occurs later in S. bombifrons than in other anurans; nonetheless, many aspects of the ossification sequence seem to be shared by a surprisingly wide range of anuran taxa.
    Additional Material: 18 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 25 (1993), S. 282-297 
    ISSN: 0886-1544
    Keywords: isoelectric focusing ; proteolysis ; cation-tubulin interactions ; microtubules ; GDP-tubulin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Limited digestion of pig brain GDP-tubulin by subtilisin was carried out in the presence of Mg2+, Mn2+, Ca2+, Zn2+, or Be2+. Isoelectric focusing, followed by SDS-PAGE, revealed characteristic divalent cation-dependent changes in the α- and β-tubulin cleavage patterns. Previous studies revealed that the β-cleavage pattern is different for heterodimers and microtubules [Lobert and Correia, 1992: Arch. Biochem. Biophys. 296:152-160]. Divalent cation effects on subtilisin digestion of tubulin indicate different classes of divalent cation binding sites. Western blot analysis locates the proteolytic zone at residue 430 or higher in both subunits for all conditions. Turbidity and electron microscopy reveal that GDP-tubulin cleaved by subtilisin in the presence of Mg2+, Ca2+, or Mn2+ forms sheets of rings. Mn2+ induces ring formation in uncleaved GDP-tubulin. Isotype-depleted tubulin was generated by the removal of class III β-tubulin using immunoaffinity chromatography. Subtilisin digestion of the depleted fraction and the purified class III β-tubulin demonstrates that cleavage occurs at three to four distinct sites. Thus, subtilisin-digested tubulin is more heterogeneous than was previously reported and the cleavage sites depend on solution conditions, divalent cations, and the state of assembly. This has important implications for experiments that utilize subtilisin-digested tubulin for studying microtubule-associated protein binding. © 1993 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 139 (1989), S. 93-99 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have reported previously that murine mammary tumor cell subpopulations isolated from one spontaneous adenocarcinoma are heterogenous in terms of prostaglandin E2 (PGE2) syntnetic capacity. We have also shown that tumor-PGE2 contributes to the ability of these cells to grow and metastasize in vivo (Fulton and Heppner: Cancer Research 45:4779-4784, 1985). In the present study, we have asked whether exogenous PGE2 has direct effects on the proliferation of these cells in vitro and if such responses can be attributed to the capacity of these cells to (1) bind PGE2 and (2) activate adenylate cyclase via the PGE2 receptor. We report that PGE2, at concentrations below 1 × 10-5 M, does not affect the proliferation rate of these cells. This unresponsiveness is not due to the absence of receptors for PGE2. However, marked heterogeneity in receptor binding and function was detected in these closely related cell lines. Two metastatic lines (66 and 410.4) have high-affinity receptors for PGE2 (average Kd = 4.3 × 10-9 M/L and 4.2 × 10-9 M/L, respectively) and similar binding capacities (4.1 × 104 and 2.9 × 104 binding sites, respectively). Two nonmetastatic lines, 410 and 67, have receptors with lower affinity (Kd = 8.3 × 10-9 M/L and 1.6 × 10-7 M/L, respectively) and binding capacities of 2.8 × 105/410 cell or 7.3 × 104/67 cell. A third nonmetastatic line (168) exhibits no specific binding. PGE2 receptor stimulation leads to elevated intracellular cAMP in lines 66, 410, and 67. Line 410.4 cells appear to have a functional lesion in the PGE2 receptor resulting in a failure to elevate cAMP in response to receptor occupancy. Adenylate cyclase can, however, be activated in these cells by cholera toxin, NaF, or forskolin. In comparison to the other cell lines, line 168 cells respond poorly to all cAMP-stimulating agents. Thus, we have found that PGE2 binding is a heterogenous property for these cells, and, in addition, we have identified an apparent uncoupling of PGE2 receptor to the adenylate cyclase system in one cell line.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 45 (1991), S. 93-100 
    ISSN: 0730-2312
    Keywords: cytoskeleton ; osteoblast ; cell-cell interaction ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: During bone development, osteoblasts form a contiguous layer along recently deposited osteoid and their morphology changes from fibroblast-like to cuboidal. In culture, similar changes occur with increased cell density. We examined the possible role of cyclic AMP in this process since cyclic AMP was reported to increase in fibroblasts with increased cell density and similar shape changes were seen in response to parathyroid hormone, which also increases cellular cyclic AMP in osteoblastic cells. Osteoblast-enriched rat calvaria cells were seeded at increasing density. The distribution between Triton X-100 extractable and nonextractable actin and myosin was estimated by polyacrylamide gel electrophoresis. Intracellular cyclic AMP was estimated by prelabeling the cellular ATP pool with 3H-adenine, followed by extraction and separation of 3H-cAMP by high-performance liquid chromatography. We found that osteoblastic cells contain about 40 pg actin and 5.3 pg myosin per cell. Around 60% of the actin and 70% of the myosin were in the nonextractable (crosslinked) form at cell densities of 10,000 to 50,000 cells per cm2. Above 50,000 cells/cm2, there was a cell density-dependent reduction in crosslinked actin and myosin and a concomitant increase in cellular cyclic AMP. A comparable rise in cyclic AMP, produced by incubation with phosphodiesterase inhibitors, and treatment with other agents that increase cyclic AMP produced a similar decrease in the level of cytoskeletal actin and myosin. Cytochalasin B treatment, through its effect on actin polymerization, produced similar changes in cell shape and cytoskeletal actin. The findings suggest that an elevation in intracellular cyclic AMP may play a role in the density-dependent changes in cell shape and microfilament organization observed in osteoblasts.
    Additional Material: 6 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 45 (1991), S. 101-111 
    ISSN: 0730-2312
    Keywords: parathyroid hormone ; cyclic AMP ; osteoblasts ; actin ; myosin ; calcium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Parathyroid hormone (PTH) alters the shape of osteoblastic cells both in vivo and in vitro. In this study, we examined the effect of PTH on cytoskeletal actin and myosin, estimated by polyacrylamide gel electrophoresis of Triton X-100 (1%) nonextractable proteins. After 2-5 minutes, PTH caused a rapid and transient decrease of 50-60% in polymerized actin and myosin associated with the Triton X-100 nonextractable cytoskeleton. Polymerized actin returned to control levels by 30 min. The PTH effect was dose-dependent with an IC50 of about 1 nM, and was partially inhibited by the (3-34) PTH antagonist. PTH caused a rapid transient rise in cyclic AMP (cAMP) in these cells that peaked at 4 min, while the nadir in cytoskeletal actin and myosin was recorded around 5 min. The intracellular calcium chelator Quin-2/AM (10 μM) also decreased cytoskeletal actin and myosin, to the same extent as did PTH (100 nM). To distinguish between cAMP elevation and Ca++ reduction as mediators of PTH action, we measured the phosphorylation of the 20 kD (Pl 4.9) myosin light chain in cells preincubated with [32P]-orthophosphate. The phosphorylation of this protein decreased within 2-3 min after PTH addition and returned to control levels after 5 min. The calcium ionophore A-23187 did not antagonize this PTH effect. Visualization of microfilaments with rhodamine-conjugated phalloidin showed that PTH altered the cytoskeleton by decreasing the number of stress fibers. These changes in the cytoskeleton paralleled changes in the shape of the cells from a spread configuration to a stellate form with retracting processes. The above findings indicate that the alteration in osteoblast shape produced by PTH involve relatively rapid and transient changes in cytoskeletal organization that appear to be mediated by cAMP.
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