Stimulation of cell division and DNA replication inPrototheca richardsiby passage through larval amphibian guts

Parasitology ◽  
1993 ◽  
Vol 107 (2) ◽  
pp. 119-124 ◽  
Author(s):  
T. J. C. Beebee ◽  
A. L.-C. Wong

SUMMARYPrototheca richardsi, an unpigmented heterotrophic alga, causes growth inhibition in amphibian larvae and has proved refractory to culturein Vitro.P. richardsireplication is dependent on regular passaging through tadpole digestive systems; uptake of thymidine by free-livingProtothecacells and incorporation into DNA are very low by comparison with leucine uptake and incorporation into protein, but DNA synthesis is detectable in cells isolated from tadpole intestines. DNA replication was elicited 6–8 h after ingestion in protothecans fed to tadpoles and subsequently re-isolated from them, providing that the tadpoles were fed subsequent to the ingestion. It appears that passaging through tadpole intestines provides an essential stimulus to maintaining an active cell division cycle inP. richardsi.

1991 ◽  
Vol 1 (2) ◽  
pp. 113-117 ◽  
Author(s):  
J. Reyes ◽  
L. F. Jiménez-García ◽  
M. A. Gonzalez ◽  
J. M. Vázquez-Ramos

AbstractWe have studied by means of cytology and autoradiography the effect of benzyladenine (BA, a synthetic cytokinin) on DNA metabolism during early maize germination.The data indicate that BA stimulates nuclear DNA replication. The doubling of the amount of nuclear DNA in BA-treated axes occurs earlier than in nontreated axes, and there is a three-fold increase in the mitotic index at 24 h of germination. These results provide further corroboration for the suggestion that the stimulation of DNA synthesis observed relates to a nuclear replicative type of synthesis. Possible mechanisms of action of BA are discussed.


1969 ◽  
Vol 41 (2) ◽  
pp. 641-645 ◽  
Author(s):  
R. A. Florentin ◽  
B. H. Choi ◽  
K. T. Lee ◽  
W. A. Thomas

1979 ◽  
Author(s):  
K. L. Kellar ◽  
B. L. Evatt ◽  
C. R. McGrath ◽  
R. B. Ramsey

Liquid cultures of bone marrow cells enriched for megakaryocytes were assayed for incorporation of 3H-thymidine (3H-TdR) into acid-precipitable cell digests to determine the effect of thrombopoietin on DNA synthesis. As previously described, thrombopoietin was prepared by ammonium sulfate fractionation of pooled plasma obtained from thrombocytopenic rabbits. A control fraction was prepared from normal rabbit plasma. The thrombopoietic activity of these fractions was determined in vivo with normal rabbits as assay animals and the rate of incorporation of 75Se-selenomethionine into newly formed platelets as an index of thrombopoietic activity of the infused material. Guinea pig megakaryocytes were purified using bovine serum albumin gradients. Bone marrow cultures containing 1.5-3.0x104 cells and 31%-71% megakaryocytes were incubated 18 h in modified Dulbecco’s MEM containing 10% of the concentrated plasma fractions from either thrombocytopenic or normal rabbits. In other control cultures, 0.9% NaCl was substituted for the plasma fractions. 3H-TdR incorporation was measured after cells were incubated for 3 h with 1 μCi/ml. The protein fraction containing thrombopoietin-stimulating activity caused a 25%-31% increase in 3H-TdR incorporation over that in cultures which were incubated with the similar fraction from normal plasma and a 29% increase over the activity in control cultures to which 0.9% NaCl had been added. These data suggest that thrombopoietin stimulates DNA synthesis in megakaryocytes and that this tecnique may be useful in assaying thrombopoietin in vitro.


1984 ◽  
Vol 4 (8) ◽  
pp. 1476-1482
Author(s):  
H Ariga

The replicating activity of several cloned DNAs containing putative origin sequences was examined in a cell-free extract that absolutely depends on simian virus 40 (SV40) T antigen promoting initiation of SV40 DNA replication in vitro. Of the three DNAs containing the human Alu family sequence (BLUR8), the origin of (Saccharomyces cerevisiae plasmid 2 micron DNA (pJD29), and the yeast autonomous replicating sequence (YRp7), only BLUR8 was active as a template. Replication in a reaction mixture with BLUR8 as a template was semiconservative and not primed by a putative RNA polymerase III transcript synthesized on the Alu family sequence in vitro. Pulse-chase experiments showed that the small-sized DNA produced in a short-term incubation was converted to full-length closed circular and open circular DNAs in alkaline sucrose gradients. DNA synthesis in extracts began in a region of the Alu family sequence and was inhibited 80% by the addition of anti-T serum. Furthermore, partially purified T antigen bound the Alu family sequence in BLUR8 by the DNA-binding immunoassay. These results suggest that SV40 T antigen recognizes the Alu family sequence, similar to the origin sequence of SV40 DNA, and initiates semiconservative DNA replication in vitro.


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.


Biochemistry ◽  
1995 ◽  
Vol 34 (24) ◽  
pp. 7913-7922 ◽  
Author(s):  
Ken Matsumoto ◽  
Masayuki Seki ◽  
Chikahide Masutani ◽  
Shusuke Tada ◽  
Takemi Enomoto ◽  
...  

1979 ◽  
Vol 150 (1) ◽  
pp. 196-201 ◽  
Author(s):  
H R MacDonald ◽  
R K Less

The requirement for DNA synthesis during the primary differentiation of cytolytic T lymphocytes (CTL) had been investigated. CTL were induced polyclonally in vitro by stimulation of normal C57BL/6 spleen cells with concanavalin A (Con A)and their cytolytic activity was tested against 51Cr-labeled target cells in the presence of Bacto Phytohemagglutinin M. With this system, CTL activity could first be detected 48 h after exposure of spleen cells to Con A. Addition of cytosine arabinoside at concentrations sufficient to reduce DNA synthesis by 95-98% in Con A-stimulated cultures did not significantly inhibit the generation of cytolytic activity on a cell-to-cell basis. These results demonstrate that derepression of the genetic information required for the expression of CTL function can occur in the absence of detectable DNA synthesis.


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