Temperature response of the cell cycle of Haplopappus gracilis in suspension culture and its significance to the G1 transition probability model

The effects of temperature on the cell cycle of Haplopappus gracilis suspension cultures were analysed by the fraction of labelled mitoses method. Sphase in these cultures shows a different temperature optimum as compared to optima derived for G₂ and mitosis. G₁ phase has a much lower Q₁₀ than the other cell cycle phases and shows no temperature optimum between 22 and 34° C. These results are discussed in relation to a transition probability model of the cell cycle proposed by Smith and Martin (Proc. Natl. Acad. Sci. USA 70, 1263–1267, 1973), in which each cell has a time independent probability of initiating the transition into another round of DNA replication and division. The implications of such a model for cell cycle analysis are discussed and a tentative model for a probabilistic transition trigger is advanced.Abbreviations FLM Fraction of labelled mitoses - T_B Total B-phase     

Chloroplast phosphoribulokinase associates with yeast phosphoriboisomerase in the presence of substrate

Pea chloroplastic phosphoribulokinase and yeast phosphoriboisomerase partition independently of one another in a two-phase polyethyleneglycol, dextran system, but apparent interaction is seen when ribose-5-phosphate is added to the two-phase system. It appears that the pea leaf of kinase recognizes yeast isomerase when it is carrying metabolite.     

Spawning of hermatypic corals in Bermuda: a pilot study

This study investigates spawning of 4 hermatypic coral species from the subtropical environment of Bermuda. Laboratory evidence of spawning behaviour is supported by synchronous field observations. Development of scleractinian planulae to postlarval stages is recorded. Diploria strigosa, D. labyrinthiformis, Montastrea annularis and M. cavernosa shed highly buoyant, pigmented eggs (300–440 µm diam.) during July to September 1986. Brief spawning periods, synchronous between conspecific colonies, were recorded for M. annularis (July and August) and M. cavernosa (August) within 1 d of the last quarter of the lunar cycle. In August, there were overlaps amongst the spawning dates of D. strigosa and the Montastrea species. Nocturnal spawning periods differed between M. annularis and M. cavernosa. This constitutes the first evidence from an Atlantic community of overlapping spawning dates amongst several faviid species, and of the accumulation of scleractinian eggs and planulae in surface slicks.     

Cell Cycling of Astrocytes and Their Precursors in Primary Cultures: A Mevalonate Requirement Identified in Late G1, but Before the G1/S Transition, Involves Polypeptides

The relationship between mevalonate and cell cycling was investigated in developing glial cells. Primary cultures of newborn rat brains were serum-depleted (0.1%, vol/vol) for 48 h on days 4-6 in vitro, then returned to 10% calf serum (time 0). After 48 h, 70-80% of the cells were glial fibrillary acidic protein (GFAP)-negative by indirect immunofluorescence; 79 +/- 7% were GFAP-positive after an additional 3 days. Serum shift-up resulted in 12 h of quiescence, and then by 20 h (S phase) in increased proportions of cells synthesizing DNA (from 15 +/- 6% to 75 +/- 4% by bromodeoxyuridine immunofluorescence at 12 h and 20 h, respectively) and rates of DNA synthesis (42 +/- 6 versus 380 +/- 32 cpm/micrograms of protein/h of [3H]thymidine uptake). Additional mevalonate (25 mM) for 30 min at 10 h reversed the inhibition of DNA synthesis apparent with mevinolin (150 microM), an inhibitor of mevalonate synthesis, present from time 0. Cycloheximide added simultaneously with mevalonate prevented this reversal of inhibition. To cause arrest at G1/S, cultures were exposed to hydroxyurea between 10 and 22 h. By 3 h after hydroxyurea removal, bromodeoxyuridine-labeled nuclei increased from 0% to 75 +/- 9%, and DNA synthesis increased 10-fold. Mevinolin failed to inhibit these increases. Thus, primary astroglial precursors stimulated to progress through the cell cycle express a mevalonate requirement in late G1, but before the G1/S transition. The effect of mevalonate was characterized further as being brief (30 min) and as requiring polypeptides.     

Alternation between attachment mechanisms by limpets in the field

The attachment mechanism used by limpets in the rocky, wave-swept intertidal zone of California was determined during high tide and low tide. The two mechanisms that limpets are known to use, suction and glue-like adhesion, were distinguished by measuring the limpets' attachment forces in shear and by staining for glue-like residues where the limpets had been attached. The results show that ≈ 73% of limpets at high tide use suction, while the rest use glue-like adhesion. Conversely, ≈ 75% of limpets at low tide use glue-like adhesion, while the rest use suction. The normal tenacity of limpets was also measured at high and low tide. The mean tenacity at high tide was significantly less than that at low tide. From these data it was estimated that the mean tenacity of glue-like adhesion is ≈ 0.23 MN·m⁻² and the mean tenacity of suction adhesion is ≈ 0.09 MN·m⁻². It is hypothesized that the cycle of alternating attachment mechanisms is linked to the limpets foraging cycles.     

Cell cycle inHelianthus tuberosus tuber tissue in relation to dormancy

Summary Observations are presented on the patterns of DNA synthesis and mitotic activity in medullary parenchyma cells excised from tubers ofHelianthus tuberosus in four different periods of dormancy. Dormancy break (activation) was induced byin vitro culture on media added with 2,4-dichlorophenoxyacetic acid. The cell cycle responsein vitro to different combinations of growth substances has also been investigated.The results show that remarkable changes in the timing of the first and second cell cycles and their phases occur with the progression of dormancy. With increasing time after tuber harvest, the following behaviours are observed: (i) a lengthening of the first cell cycle, chiefly due to a lengthening of the G₂ phase (G₂ is absent at the beginning of dormancy) and an increase in the time interval between the start of thein vitro culture and the onset of the first mitotic wave; (ii) an increased duration of the S phase; (iii) a remarkable reduction in the cell synchrony.These behaviours, as indicated also by their comparison with thein vitro response of the cell cycle to different hormonal treatments, seem to depend on the physiological status of the tubers at the time of explant. It is concluded that the analysis of the cell cycle is an useful tool for understanding some aspects of such a complex physiological situation as dormancy.Istituto di Mutagenesi e Differenziamento del C.N.R., Pisa, Italy, publication no. 321.     

Alternative Pathways of Glucose Utilization in Brain; Changes in the Pattern of Glucose Utilization in Brain Resulting from Treatment of Rats with 6-Aminonicotinamide

The effect of 6-aminonicotinamide (6AN) treatment on the activities of alternative pathways of glucose metabolism in 20-day-old rat brain was evaluated by measurements of yields of 14CO2 from glucose labeled with 14C on carbons 1, 2, 3 + 4, or 6 and uniformly labeled glucose, and from the incorporation of 14C from specifically labeled glucose into lipids by brain slices from cerebral hemispheres and cerebellum. At the highest dose of 6AN used (35 mg/kg body weight) there was a significant decrease in the 14CO2 yields via the pentose phosphate pathway, the glycolytic route, tricarboxylic acid (TCA) cycle, and via the glutamate-gamma-aminobutyric acid pathway. Giving a graded series of doses (20-35 mg 6AN/kg body weight) revealed a hierarchy of responses in which the pentose phosphate pathway, lactate, glyceride-glycerol, and fatty acid formation were most sensitive, followed, in sequence, by the pyruvate dehydrogenase reaction, the glutamate-gamma-aminobutyrate route and, finally, the TCA cycle. The nature of the blocks in the various pathways was examined by the use of metabolite profiles.     

Synchronization of 9L rat brain tumor cells by centrifugal elutriation

Asynchronous 9L cells were separated into relatively homogeneously-sized populations using centrifugal elutriation with both a conventional collection method and a long collection method. A substantial increase in the homogeneity of the volume distributions and in the degree of synchrony of the separated fractions was obtained using the long collection method. Autoradiographic data indicated that fractions containing ≥97% G₁ cells, ≥80% S cells, and 70–75% G₂ cells could be routinely recovered with this procedure. Recovery in these fractions varied from 5 to 8% of the total number of cells elutriated. The colony forming efficiency (CFE) of cells from fractions representing each phase of the cell cycle was a constant 60–70%, which was comparable to the 60–80% usually found for asynchronous 9L cells. The percentage of cells in the G₁, S, and G₂ phases in the elutriated fractions was more accurately determined from the volume distribution than from computer fits of the DNA histogram obtained from flow cytometry. In general, the degree of synchrony was related to the coefficient of variation (CV) of the volume distributions of the elutriated fractions. The CV was about 14% for all elutriated fractions. When the ≥97% G₁ population was allowed to progress to S and G₂, the CVs were about 17 and 20.2%, respectively. Thus, the best nonperturbing method for obtaining synchronous 9L cells in the S or G₂ phases was direct elutriation with the long collection method.