An Ultrastructural and Radio-Autographic Study of the Evolution of the Interphase Nucleus in Plant Meristematic Cells (Allium Porrum)

1974 ◽  
Vol 14 (2) ◽  
pp. 263-287
Author(s):  
J. G. LAFONTAINE ◽  
A. LORD

Radioautography under both light and electron microscopy was exploited to investigate the structural changes of the chromatin reticulum which characterizes the interphase nucleus of a number of plants. Allium porrum meristematic plant cells were used for this purpose. In this species, the telophase chromosomes uncoil into dense strands which, during the G1 period, gradually give rise to a coarse reticulum. There then follows an extensive unravelling of portions of these strands, and high-resolution radioautography reveals that labelling with tritiated thymidine predominantly occurs over zones of the nucleus consisting of diffuse fine fibrillar material. As the S-period progresses, a chromatin reticulum reappears throughout the nuclear cavity, the tortuous strands being approximately 0.25 µm in diameter. Most of the radioautographic grains still remain over the light nucleoplasmic areas but a number of these are now located on the outermost portion of the dense chromatin profiles. By the end of the S-period, the chromatin strands are slightly thicker (ca. 0.3 µm) and form a looser reticulum. Labelling has decreased noticeably in nuclei of that period, the radioautographic grains being grouped into clusters resting over more or less spherical regions of the chromatin reticulum. Judging from their localization at the surface of the nucleolus or close to the nuclear envelope, these structures correspond to chromocentres. The additional interesting finding that such nuclear structures appear much less compactly organized strongly suggests that chromocentres undergo important conformational modifications during duplication of their DNA.

1963 ◽  
Vol 16 (1) ◽  
pp. 29-51 ◽  
Author(s):  
Elizabeth D. Hay ◽  
J. P. Revel

In the present investigation, the sites of deoxyribonucleic acid (DNA) synthesis and the fate of labeled deoxyribonucleoprotein (DNP) were studied in autoradiographs of ultrathin sections viewed with the electron microscope. Tritiated thymidine was employed as a label for DNA in the nuclei of proliferating cells of regenerating salamander limbs. In the autoradiographic method reported here, dilute NaOH was used to remove the gelatin of the emulsion after exposure and development. The exposed silver grains are not displaced by this treatment and the resolution of fine structure in the underlying section is greatly improved. Our observations suggest that the DNP component is a meshwork of interconnected filaments 50 to 75 A in diameter, which may be cross-linked to form what Frey-Wyssling would term a "reticular gel." The filamentous DNP meshwork is dispersed throughout the interphase nucleus during DNA synthesis, whereas in chromosomes, which are relatively inert metabolically, the meshwork is denser and is aggregated into compact masses. Dense chromatin centers in interphase nuclei are similar in fine structure to chromosomes and are also inert with respect to DNA synthesis. In the Discussion, the structure of the filamentous meshwork in chromatin is compared with that in chromosomes, and speculations are made as to the functional significance of the variations in DNP fine structure observed.


1976 ◽  
Vol 21 (1) ◽  
pp. 193-207
Author(s):  
A. Lord ◽  
J.G. Lafontaine

In Raphanus sativus, the mitotic chromosomes are quite short and, on reaching the cell poles, soon undergo extensive unravelling. By late telophase and early interphase, only a few chromosome segments, believed to correspond to the centromeric regions, are still visible in the form of chromocentres closely associated with the nuclear envelope. Although interphase nuclei show little internal structural differentiation, high-resolution radioautography has permitted us to establish which of them have reached the early, mid and late S periods. In early S nuclei, only the nucleolus and the euchromatin which pervades the nuclear cavity become labelled. By the mid S-period, the diffuse chromatin and nucleolus incorporate less thymidine and DNA synthesis is initiated within the peripheral chromocentres. Subsequently, the radioautographic grains become restricted to the chromocentres. The finding that certain late S nuclei exhibit loosely organized chromocentres strongly suggests that these heterochromatic chromosome segments undergo important conformational modifications during DNA replication. Finally, the presence of radioautographic grains over the lacunar regions of the nucleolus in early and mid S nuclei demonstrates that intranucleolar DNA replicates during the earlier portion of the S-period.


1973 ◽  
Vol 12 (2) ◽  
pp. 369-383
Author(s):  
J.-G. LAFONTAINE ◽  
A. LORD

The interphase nucleolus in Allium porrum meristematic cells is characterized by the presence of 1-4 dense fibrillar zones of rather complex organization. Each such zone appears to consist essentially of a convoluted, evacuolated, filamentous structure approximately 1.5 µm in diameter. At the ultrastructural level, these structures exhibit an intricate array of lacunar spaces each of which is surrounded by a dense coating. These lacunae are filled with a loose fibrillar material and the largest ones sometimes also show a dense central core. In appropriate preparations, certain of the peripherally located lacunae are found to be continuous with segments of chromosomes. High-resolution radioautography reveals, moreover, that DNA is present within both the dense and lighter portions of the nucleolar loops. These observations add further support to the hypothesis that the convoluted filamentous structures in question correspond to loops of chromosomal origin and are thus related to the nucleolar organizer.


IAWA Journal ◽  
1990 ◽  
Vol 11 (4) ◽  
pp. 413-420 ◽  
Author(s):  
Uwe Schmitt ◽  
Walter Liese

Reactions in the xylem parenchyma of Betula pendula Roth following wounding in late spring have been investigated by light and electron microscopy. Structural changes in contact parenchyma cells are described in relation to the formation of plugs in vessels and fibres. Swelling of the Protective Layer (PL) with a simultaneous loss in electron density appears first. Fibrillar material is then synthesised, which accumulates outside the cytoplasm between the plasmalemma and the modified PL. The PL now consists of loosely packed fibrils with a structure similar to the accumulating fibrillar material. After extrusion of fibrils through the pit membrane into the lumina of adjacent vessels a membranous layer on the scalariform perforation plates as well as plugs are formed; the latter are also built up in fibres. These reactions spread axially less extensively in cells near the cambium than in the more centrally located ones.


1973 ◽  
Vol 13 (1) ◽  
pp. 97-119
Author(s):  
A. LORD ◽  
J. G. LAFONTAINE

In the present work the effects of an intermediate dose (20µg/ml) of ethidium bromide on nuclear structures of meristematic plant cells (Raphanus sativus) have been studied under both light and electron microscopy. Radioautography was also exploited to assess the action of this drug on the incorporation activity of nucleic acid precursors. After a 30-min treatment with ethidium bromide, no noticeable ultrastructural modifications are detected but the incorporation of both thymidine and uridine is already markedly impaired. As treatment is extended to 1 h, the numerous light lacunae characterizing the fibrillar nucleolar zones in normal cells transform into elongated areas which are often found to extend to the surface of this organelle and are continuous with perinucleolar chromatin masses. After 2 h, many small bodies consisting of granular material appear in the nucleoplasm. The nucleolus gradually loses most of its granular component and, eventually, becomes predominantly fibrillar in texture; in many cells it breaks up into a number of irregular masses. Throughout various stages of the experiment, certain cells show small spherical nucleoli characterized by the presence on their surface of one or two conspicuous reticulate masses which are heavily labelled with thymidine and, consequently, consist mostly of chromatin. In view of the fact that ethidium bromide has been observed in this study to induce a certain degree of condensation of chromatin within the nuclear cavity, it is surmised that this drug also brings about migration of intranucleolar DNA to the surface of this organelle.


2021 ◽  
Vol 26 (5) ◽  
pp. 30-38
Author(s):  
E. A. Savchuk ◽  
E. P. Golubinskaya ◽  
T. N. Shcherbinina ◽  
G. Yu. Voronin ◽  
E. O. Savchuk ◽  
...  

The article presents an analysis of the literature and a clinical case of a rare disease from the group of diseases of small vessels — cerebral autosomal dominant arteriopathy with subcortical infarction and leukoencephalopathy (CADASIL).It is based on the deposition of osmiophilic granulation material in vessels of small and medium caliber. A mutation in the NOTCH3 gene on chromosome 19p13 leads to significant structural changes in the walls of small arteries due to impaired differentiation and maturation of smooth muscle cells.CADASIL is characterized by four key symptoms: migraines, recurrent ischemic strokes, mental disorders, and cognitive decline. The clinical case study is presented from the standpoint of a multidisciplinary patient-oriented approach of joint work of neurologists and morphologists. On the basis of clinical and laboratory criteria, a probable diagnosis was made. To confirm it, a muscle biopsy was performed (a musculocutaneous flap from the inner surface of the thighs and forearms), in order to conduct light and electron microscopy. The details of the results of the morphological study, which made it possible to verify the patient’s diagnosis, are presented. Differential diagnostic judgments are presented and recommendations for genetic studies in the family, prognosis and treatment of the patient are given.


2011 ◽  
Vol 300 (2) ◽  
pp. H423-H430 ◽  
Author(s):  
Yasumi Uchida ◽  
Yasuto Uchida ◽  
Akimasa Matsuyama ◽  
Atsushi Koga ◽  
Yuko Maezawa ◽  
...  

Although there are a number of studies on vasospastic angina, the structural changes at the cellular level that occur in the coronary arterial wall during spasm are not well known. Coronary spasm was induced by brushing the coronary adventitia in nine anesthetized beagles, and structural changes in the spastic coronary segments were examined by light and electron microscopy, making comparisons with the adjacent nonspastic segments. The % diameter stenosis of the spastic segments as measured angiographically was 79.4 ± 12% (mean ± SD). Light microscopic changes in the spastic and nonspastic segments were as follows: medial thickness 1,512 vs. 392 μm ( P < 0.0001) and % diameter and % area stenoses of spastic segment 81.0% and 96.5%, respectively, indicating that spasm was induced by medial thickening. Circular smooth muscle cells (SMCs) in the media were arranged in parallel with the internal (IEL) and external (EEL) elastic lamina in nonspastic segments but radially rearranged in spastic segments. SMCs were classified by their patterns of connection to IEL into six types by electron microscopy. Of these, three contracted and pulled the IEL toward the EEL, causing folding of the IEL and waving of EEL resulting in thickening of the media and narrowing of the lumen. We conclude that coronary spasm was elicited by radial rearrangement of the medial SMCs due to their own contraction and resultant medial thickening and folding of IEL, creating a piston effect to narrow the lumen, i.e., spasm.


1997 ◽  
Vol 110 (16) ◽  
pp. 1919-1934
Author(s):  
M. Murate ◽  
Y. Kishimoto ◽  
T. Sugiyama ◽  
T. Fujisawa ◽  
H. Takahashi-Iwanaga ◽  
...  

Hydra tissue consists of the ectodermal and the endodermal layers. When the two layers were separated by procaine treatment and then recombined, the ectodermal epithelial cells spread as a single cell layer over the endoderm as in epiboly in vertebrate embryogenesis, and the resultant spherical structure subsequently regenerated into a complete hydra. In this study, light and electron microscopy were used to examine the structural changes which took place in the cells and tissue during this epibolic ectodermal spreading process. Within a few hours after tissue recombination, the endoderm underwent dramatic changes; it lost its epithelial sheet organization, and turned into a mass of irregularly shaped cells without the apical-basal cell polarity initially present. In contrast, the ectoderm maintained its basic epithelial sheet organization as it spread over the endoderm. Later, the endodermal epithelial cells reorganized themselves into a single-layered epithelial sheet underneath the spreading ectodermal layer. The resultant spherical structure consisted of a single layer of ectodermal epithelial cells outside, a single layer of endodermal epithelial cells inside, and an empty cavity in the center as in normal hydra tissue. This structure regenerated into hydra in the following days. These and other observations demonstrate that the two-layered epithelial sheet organization is highly dynamic, and that its stability is maintained by strong interactions between the two layers in normal hydra. It is suggested that this dynamic nature of the hydra tissue, particularly the high plasticity of the endodermal epithelial sheet organization, may be an important element for the high regenerative capacity of this organism.


1993 ◽  
Vol 30 (5) ◽  
pp. 401-409 ◽  
Author(s):  
G. Abebe ◽  
M. K. Shaw ◽  
R. M. Eley

The pituitary glands of seven Boran cattle ( Bos indicus), five infected with a clone of Trypanosoma congolense IL 1180 (ILNat 3.1) transmitted by Glossina morsitans centralis and two uninfected controls, were examined by light and electron microscopy 43 (experiment 2) or 56 (experiment 1) days after fly challenge. The three cattle used in the first experiment included a 15-month-old female (No. 1), a 24–month-old female (No. 2), and a 21–month-old male (No. 3) as a control. In the second experiment, four cattle were used: two females (Nos. 4, 5) and one male (No. 6), all between 15 and 24 months of age, and one female control (No. 7) of similar age. In all the infected animals, dilation of both the sinusoids and microvasculature was apparent, as was an increase in the thickness of the extracellular matrix between the pituitary lobules. Trypanosomes were found in the microvasculature of the adenohypophysis and neurohypophysis in all the infected animals. Focal degenerative changes were seen in the adenohypophyseal section of glands from the infected animals euthanatized 56 days post-infection. These degenerative structural changes were confined to the somatotrophs cells. The possible role that trypanosomes in the microvasculature may play in inducing pituitary damage and dysfunction is discussed.


1961 ◽  
Vol 201 (1) ◽  
pp. 213-216 ◽  
Author(s):  
R. J. M. Fry ◽  
S. Lesher ◽  
H. I. Kohn

Epithelial cell renewal in the jejunum of CAF1 mice was studied by observing the time for cells labeled with tritiated thymidine (1.5–2.0 µc/g body wt.) to pass from the crypt to the extrusion zone at the tip of the villus. Mice of three ages were studied: 93, 372, and 940 days. The total observed transit times were 44, 52, and 53 hr, respectively. The increase with age was the result of increase in both the crypt and villus transit times. Structural changes could not be made to account for the differences. The increase may be the result of a decreased rate of cell production in the crypts.


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