Cytoplasmic DNA Synthesis at Meiotic Prophase in Lilium henryi

1979 ◽  
Vol 27 (3) ◽  
pp. 273 ◽  
Author(s):  
DR Smyth ◽  
T Shaw

Extensive cytoplasmic DNA synthesis has been discovered at meiotic prophase in Lilium henryi. Explanted microsporocytes were cultured in medium containing I3H]thymidine. Light microscope autoradiography revealed many small clusters of grains (< 3 μm) in the cytoplasm of premeiotic and pachytene cells. There were about 10 to 20 clusters per cell in each section. Most of the cyto- plasmic label was extracted by hot hydrochloric acid and DNase I, but not by RNase A or pronase. Thus the grains reflect DNA synthesis, and not incorporation of products of thymidine catabolism which is extensive in this tissue. These localized centres of DNA synthesis, previously unreported, might result from mitochondrial or plastid replication, or from amplification of excised chromosomal DNA.

1964 ◽  
Vol 22 (3) ◽  
pp. 505-513 ◽  
Author(s):  
D. R. Wolstenholme ◽  
W. Plaut

The application of electron microscope autoradiography to Amoeba proteus cells labeled with tritiated thymidine has permitted the identification of morphologically distinct particles in the cytoplasm as the sites of incorporated DNA precursor. The particles correspond to those previously described from light microscope studies, with respect to both H3Tdr incorporation and distribution in centrifugally stratified amoebae. Ingested bacteria differ from the particles, in morphology as well as in the absence of associated label. Attempts to introduce a normal particle labeling pattern by incubating amoebae with labeled sediment derived from used amoeba medium failed. The resultant conclusion, that the particles are maintained in the amoeba by self-duplication, is supported by the presence of particles in configurations suggestive of division.


1983 ◽  
Vol 29 (2) ◽  
pp. 254-260 ◽  
Author(s):  
Malgorzata Kloc ◽  
Peter E. Lee

Viral-specific DNA synthesis of Tipula iridescent virus (TIV) was not affected in Estigmene acrea cells which were continuously exposed to 300 μg/mL hydroxyurea (HU) as detected by light microscope autoradiography. Electron microscopy of such cells showed viroplasmic centres with virions in various stages of development. In nor mal cells similarly exposed to HU, nuclear DNA synthesis was reduced by 70–80%.


1962 ◽  
Vol 15 (3) ◽  
pp. 535-540 ◽  
Author(s):  
M. Rabinovitch ◽  
W. Plaut

Nucleic acid-containing particles in the cytoplasm of Amoeba proteus (cf. reference 1) were counted after acridine orange staining. The number of particles per ameba was found to be correlated with cell age and size. Fresh daughters had a mean particle number of 5400, whereas predivision amebae contained around 11,000 particles. Amebae from two other strains contained similar particles. The particles were found to be clustered in fasted cells and redispersed after feeding. A marked increase in the particle population was noted in anucleate fragments. These results, together with those previously presented, suggest that the particles multiply intracellularly. Their nature and their relationship to previous work on nucleic acid labeling in Amoeba are discussed.


1980 ◽  
Vol 44 (1) ◽  
pp. 375-394
Author(s):  
N.N. Bobyleva ◽  
B.N. Kudrjavtsev ◽  
I.B. Raikov

The DNA content of isolated micronuclei, differentiating macronuclei (macronuclear Anlagen), and adult macronuclei of Loxodes magnus was measured cytofluorimetrically in preparations stained with a Schiff-type reagent, auramine-SO2, following hydrochloric acid hydrolysis. The DNA content of the youngest macronuclear Anlagen proved to be the same as that of telophasic micronuclei (2 c). The Anlagen thus differentiate from micronuclei which are still in G1. The quantity of DNA in the macronuclear Anlagen thereafter rises to the 4-c level, simultaneously with DNA replication in the micronuclei which immediately follows mitosis. In non-dividing animals most micronuclei are already in G2. Adult macronuclei here contain on average 1.5 times more DNA than the micronuclei; their DNA content is about 5–6 c (in some individual nuclei, up to 10 c). These data are consistent with autoradiographic evidence indicating a weak DNA synthesis in the macronuclei of Loxodes and make likely the existence of partial DNA replication (e.g. gene amplification) in the macronuclei. The DNA content of adult macronuclei isolated from dividing animals proved to be significantly smaller than that of macronuclei isolated from non-dividing specimens of the same clone. In 3 clones studied, the former value amounted on average to 71–79, 78 and 95% of the latter, respectively. This drop of DNA content cannot be explained by ‘dilution’ of the old macronuclei with newly formed ones. The quantity of DNA in adult macronuclei thus seems to undergo cyclical changes correlated with cytokinesis, despite the fact that, in Loxodes magnus, the macronuclei themselves never divide and are simply segregated at every cell division. The macronuclei of Loxodes can be termed paradiploid or hyperdiploid.


1962 ◽  
Vol 15 (3) ◽  
pp. 525-534 ◽  
Author(s):  
M. Rabinovitch ◽  
W. Plaut

The incorporation of tritiated thymidine in Amoeba proteus was reinvestigated in order to see if it could be associated with microscopically detectable structures. Staining experiments with basic dyes, including the fluorochrome acridine orange, revealed the presence of large numbers of 0.3 to 0.5 µ particles in the cytoplasm of all cells studied. The effect of nuclease digestion on the dye affinity of the particles suggests that they contain DNA as well as RNA. Centrifugation of living cells at 10,000 g leads to the sedimentation of the particles in the centrifugal third of the ameba near the nucleus. Analysis of centrifuged cells which had been incubated with H3-thymidine showed a very high degree of correlation between the location of the nucleic acid-containing granules and that of acid-insoluble, deoxyribonuclease-sensitive labeled molecules and leads to the conclusion that cytoplasmic DNA synthesis in Amoeba proteus occurs in association with these particles.


Development ◽  
1971 ◽  
Vol 26 (2) ◽  
pp. 271-283
Author(s):  
Y. C. Kong ◽  
I. F. Lau ◽  
W. L. Lam ◽  
C. M. Choy

Mature Megalobatrachus oocytes contain 43 µg DNA per oocyte, as compared with 250 pg DNA in a hepatocyte of the same animal. Megalobatrachus oocytes respond to CdR treatment by an increased incorporation of [3H]lysine into basic proteins associated with ooplasmic particles, with an optimal CdR concentration at 2 mM. The nucleolus is the most active site of [3H]lysine incorporation. It is suggested that CdR-stimulated basic protein synthesis is a common biochemical event during amphibian oogenesis. The dose response to CdR treatment may be a function of the c-DNA content or c-DNA synthesis potential in the ooplasm.


1976 ◽  
Vol 22 (3) ◽  
pp. 521-530
Author(s):  
I. Minassian ◽  
L.G. Bell

Light- and electron-microscope autoradiography have been used to follow the incorporation of [3H]thymidine at different stages during the interphase of synchronously growing populations of Amoeba proteus. Two main patterns were found for tritiated thymidine incorporation, i.e. DNA synthesis. The major incorporation was in the central region of the nucleus, but a lesser degree of incorporation occurred in the nucleolar region. The bulk of this nucleolar DNA was found to be late replicating, i.e. it replicated during the G2 phase.


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