The nucleolus and meiosis during microsporogenesis in Endymion non-scriptus (L.)

1977 ◽  
Vol 25 (1) ◽  
pp. 111-123
Author(s):  
B.T. Luck ◽  
E.G. Jordan

Stages of meiosis from the bluebell Endymion non-scriptus (L.) were studied by electron microscopy. The segregated components of the nucleolus at meiotic prophase underwent fragmentation and dissolution at pachytene-diplotene. Nucleoli were absent during both meiotic divisions and reformed on the nucleolus organizer into a fibrillar mass from scattered fibrillar components at the dyad and tetrad stages. Ti is argued that the fibrillar region shows continuity through nuclear division though undergoing structural transformations in the process. Nucleolar reformation occurs on condensed nucleolus organizers. Processing of the ribosomal precursors and the resumption of RNA synthesis is discussed in relation to the dispersal of the nucleolus organizer into the fibrillar region of the reformed nucleolus.

1972 ◽  
Vol 11 (3) ◽  
pp. 689-697
Author(s):  
H. M. KRIDER ◽  
W. PLAUT

The influence of conditions resulting in bobbed phenotypes on nucleolar RNA synthesis and the formation of constrictions at nucleolus organizers was examined in larval tissues of Drosophila melanogaster. By means of [3H]uridine incorporation and autoradiographic analysis, a mutation at the bobbed locus was shown to limit the rate of nucleolar RNA synthesis in salivary glands of XO larvae. The formation of constrictions at the organizer sites of a 4-nucleolus-organizer stock was monitored in dividing neuroblast cells stained with acridine orange. Loss of the ribosomal cistrons had been reported by other workers when such stocks were maintained for several generations. In the first generation in our work, constrictions were visible at only 2 of the 4 nucleolus organizers. This situation persisted until the fifth generation, when constrictions appeared at all 4 of the organizer sites. An increase in the rate of nucleolar RNA synthesis in the salivary glands was temporally correlated with the appearance of the extra constrictions. We interpret these observations to mean that 2 of the organizers of the 4-nucleolus-organizer stock were caused to function through the loss of ribosomal RNA cistrons; thus the functional status of an organizer would appear to be subject to control.


1972 ◽  
Vol 11 (3) ◽  
pp. 675-687
Author(s):  
H. M. KRIDER ◽  
W. PLAUT

Nucleolar RNA synthesis in salivary glands from XX, XY, and XO larvae having different numbers of nucleolus organizers was examined autoradiographically following incubation of the tissues in [3H]uridine. In addition, the presence or absence of secondary constrictions was monitored in neuroblast preparations using acridine orange staining. It was observed that: (1) The rate of nucleolar [3H]uridine incorporation is independent of the number of nucleolus organizers in the cell; (2) nucleolar incorporation in XO tissues is elevated relative to that observed in XX or XY material; and (3) where the number of organizers is in excess of the wild type (2), secondary constrictions form at only 2 of the nucleolus organizer sites. From these and related observations we suggest that there are 2 forms of transcriptional control for nucleolar RNA synthesis. One acts as an on-off control, influencing the formation of secondary constrictions at the nucleolus organizers. The other form modulates rates of transcription at organizers where constrictions have been formed.


Author(s):  
Jean-Luc Rouvière ◽  
Alain Bourret

The possible structural transformations during the sample preparations and the sample observations are important issues in electron microscopy. Several publications of High Resolution Electron Microscopy (HREM) have reported that structural transformations and evaporation of the thin parts of a specimen could happen in the microscope. Diffusion and preferential etchings could also occur during the sample preparation.Here we report a structural transformation of a germanium Σ=13 (510) [001] tilt grain boundary that occurred in a medium-voltage electron microscopy (JEOL 400KV).Among the different (001) tilt grain boundaries whose atomic structures were entirely determined by High Resolution Electron Microscopy (Σ = 5(310), Σ = 13 (320), Σ = 13 (510), Σ = 65 (1130), Σ = 25 (710) and Σ = 41 (910), the Σ = 13 (510) interface is the most interesting. It exhibits two kinds of structures. One of them, the M-structure, has tetracoordinated covalent bonds and is periodic (fig. 1). The other, the U-structure, is also tetracoordinated but is not strictly periodic (fig. 2). It is composed of a periodically repeated constant part that separates variable cores where some atoms can have several stable positions. The M-structure has a mirror glide symmetry. At Scherzer defocus, its HREM images have characteristic groups of three big white dots that are distributed on alternatively facing right and left arcs (fig. 1). The (001) projection of the U-structure has an apparent mirror symmetry, the portions of good coincidence zones (“perfect crystal structure”) regularly separate the variable cores regions (fig. 2).


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Goran Kokic ◽  
Hauke S. Hillen ◽  
Dimitry Tegunov ◽  
Christian Dienemann ◽  
Florian Seitz ◽  
...  

AbstractRemdesivir is the only FDA-approved drug for the treatment of COVID-19 patients. The active form of remdesivir acts as a nucleoside analog and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses including SARS-CoV-2. Remdesivir is incorporated by the RdRp into the growing RNA product and allows for addition of three more nucleotides before RNA synthesis stalls. Here we use synthetic RNA chemistry, biochemistry and cryo-electron microscopy to establish the molecular mechanism of remdesivir-induced RdRp stalling. We show that addition of the fourth nucleotide following remdesivir incorporation into the RNA product is impaired by a barrier to further RNA translocation. This translocation barrier causes retention of the RNA 3ʹ-nucleotide in the substrate-binding site of the RdRp and interferes with entry of the next nucleoside triphosphate, thereby stalling RdRp. In the structure of the remdesivir-stalled state, the 3ʹ-nucleotide of the RNA product is matched and located with the template base in the active center, and this may impair proofreading by the viral 3ʹ-exonuclease. These mechanistic insights should facilitate the quest for improved antivirals that target coronavirus replication.


1964 ◽  
Vol 23 (1) ◽  
pp. 63-78 ◽  
Author(s):  
James R. Coleman ◽  
Montrose J. Moses

The indium trichloride method of Watson and Aldridge (38) for staining nucleic acids for electron microscopy was employed to study the relationship of DNA to the structure of the synaptinemal complex in meiotic prophase chromosomes of the domestic rooster. The selectivity of the method was demonstrated in untreated and DNase-digested testis material by comparing the distribution of indium staining in the electron microscope to Feulgen staining and ultraviolet absorption in thicker sections seen with the light microscope. Following staining by indium, DNA was found mainly in the microfibril component of the synaptinemal complex. When DNA was known to have been removed from aldehyde-fixed material by digestion with DNase, indium stainability was also lost. However, staining of the digested material with non-selective heavy metal techniques demonstrated the presence of material other than DNA in the microfibrils and showed that little alteration in appearance of the chromosome resulted from DNA removal. The two dense lateral axial elements of the synaptinemal complex, but not the central one to any extent, also contained DNA, together with non-DNA material.


1958 ◽  
Vol 4 (3) ◽  
pp. 267-274 ◽  
Author(s):  
August Ruthmann

Histochemical procedures for the demonstration of RNA have shown the presence of intensely basophilic bodies in the cytoplasm of spermatocytes of the crayfish, Cambarus virilis. The staining of thick sections, cut alternately with thin sections for electron microscopy, has permitted identification of the basophilic bodies with two types of lamellar systems. One of these, a set of straight annulate lamellae, is restricted to meiotic prophase. The second type of lamellar systems has been found from late prophase to early spermatid stages. It consists of an ellipsoidal lamellar set which intersects a number of straight lamellae. Within the region of intersection, the ellipsoidal lamellae break up into an array of small tubules of about 150 A diameter. The term tubulate lamellar system was chosen to designate this type of lamellar complex. Small RNA-containing granules could not be detected in annulate lamellar systems. While there are a few granules in the marginal regions of the tubulate lamellar system, their distribution cannot be responsible for the basophilia which is intense within all regions of the lamellar body.


1972 ◽  
Vol 53 (2) ◽  
pp. 483-493 ◽  
Author(s):  
Laura L. Tres ◽  
A. L. Kierszenbaum ◽  
C. J. Tandler

Earlier reports indicated the presence of significant amounts of inorganic salts in the nucleus. In the present study the possibility that this might be related to the transcription process was tested on seminiferous epithelium of the adult mouse, using potassium pyroantimonate as a fixative. The results indicated that a correlation exists between the inorganic cations comprising the pyroantimonate-precipitable fraction and the RNA synthetic activity. During meiotic prophase an accumulation of cation-antimonate precipitates occurs dispersed through the middle pachytene nuclei, the stage in which RNA synthesis reaches a maximum. At other stages (zygotene to diplotene), where RNA synthesis falls to a low level, that pattern is not seen; cation-antimonate deposits are restricted to a few masses in areas apparently free of chromatin. The condensed sex chromosomes, the heterochromatin of the "basal knobs," the axial elements, and the synaptonemal complexes are devoid of antimonate deposits during the meiotic prophase. The Sertoli cells, active in RNA synthesis in both nucleoplasm and nucleolus, show cation-antimonate deposits at these sites. In the nucleoplasm some "patches" of precipitates appear coincident with clusters of interchromatin granules; in the nucleolus the inorganic cations are mainly located in the fibrillar and/or amorphous areas, whereas relatively few are shown by the granular component. The condensed chromatin bodies associated with the nucleolus were always free of antimonate precipitates. It is suggested that the observed sites of inorganic cation accumulation within the nucleus may at least partially indicate the presence of RNA polymerases, the activity of which is dependent on divalent cations.


1973 ◽  
Vol 58 (1) ◽  
pp. 54-63 ◽  
Author(s):  
David M. Phillips ◽  
Stephanie Gordon Phillips

The reconstruction of the nucleolus after mitosis was analyzed by electron microscopy in cultured mammalian (L929) cells in which nucleolar RNA synthesis was inhibited for a 3 h period either after or before mitosis. When synchronized mitotic cells were plated into a concentration of actinomycin D sufficient to block nucleolar RNA synthesis preferentially, nucleoli were formed at telophase as usual. 3 h after mitosis, these nucleoli had fibrillar and particulate components and possessed the segregated appearance characteristic of nucleoli of actinomycin D-treated cells. Cells in which actinomycin D was present for the last 3 h preceding mitosis did not form nucleoli by 3 h after mitosis though small fibrillar prenucleolar bodies were detectable at this time. These bodies subsequently grew in size and eventually acquired a particulate component. It took about a full cell cycle before nucleoli of these cells were completely normal in appearance. Thus, nucleolar RNA synthesis after mitosis is not necessary for organization of nucleoli after mitosis. However, inhibition of nucleolar RNA synthesis before mitosis renders the cell incapable of forming nucleoli immediately after mitosis. If cells are permitted to resume RNA synthesis after mitosis, they eventually regain nucleoli of normal morphology.


Sign in / Sign up

Export Citation Format

Share Document