Cell cycle analysis of facial mesenchyme in the chick embryo

Development ◽  
1984 ◽  
Vol 81 (1) ◽  
pp. 49-59
Author(s):  
Robert Minkoff

Cell cycle parameters were analysed in mesenchyme of the maxillary process and the roof of the stomodeum in the chick embryo from stages 19 through 28. The generation times at stages 24–26 were determined by pulse labelling of embryos with [3H]thymidine, followed by labelled mitosis counts and construction and analysis of percent-labelled mitosis curves employing computer-assisted curve-fitting techniques. The median generation time was approximately 10·6 h in the maxillary process, and 16 h in the roof of the stomodeum; corresponding values for mean generation times were approximately 12·0 and 18·2 h, respectively. Median values for transit times of G1, S, and G2 were 2·0, 5·4, and 2·5 h in the maxillary process and 5·2, 6·7, and 2·7 h in the roof of the stomodeum. The distribution of generation times of cells in the roof of the stomodeum, however, appeared to be more heterogeneous than those of cells in the maxillary process. The percentage of cells which continue to cycle rapidly (i.e. the ‘growth fraction’) was determined by repeated-labelling experiments with [3H]thymidine in chick embryos from stages 19 through 28. Cumulative labelling of mesenchymal cells in both the maxillary process and roof of the stomodeum approached 100 % at stage 19 but dropped markedly from stage 19 to 25 declining to approximately 60–70 % in the maxillary process, and to 30 % in the roof of the stomodeum. The decline in cell proliferation rates for these regions, determined in previous studies with labelling indices, appears to be a result of the removal of cells from rapidly cycling cell populations into subpopulations which are cycling more slowly and possibly into subpopulations which have become quiescent; the difference in growth rates between these regions could be attributed to the time of appearance and the size of these emerging slow cycling or quiescent subpopulations.

Development ◽  
1974 ◽  
Vol 32 (3) ◽  
pp. 697-705
Author(s):  
Doris B. Wilson ◽  
E. M. Center

The cell cycle of mesencephalic ventricular cells was studied by means of tritiated thymidine radioautography during normal and abnormal development in the looptail (Lp ) mutant mouse. The total generation time, DNA-synthetic (S), premitotic (G2), mitotic (M), and postmitotic (G1) periods were compared in looptail homozygotes (Lp/Lp ) which exhibit neural dysraphism and in their normal littermates (+ / +) at 10 and 11 days ’ gestation. Both normal and abnormal embryos showed a chronological lengthening of the generation time between the 10th and 11th day. However, the generation time in the 10-day abnormal brains was 4·5 h longer than that in normal littermates, and the difference was the result of an increase mainly in the M and G1 periods. At 11 days of gestation the generation time in the abnormal brains increased by 50 h over that of the normal brains. Since the cell cycle was actually prolonged in the defective brains, the increased numbers of mitotic figures which characterize the looptail homozygote brain during early development appear to reflect the lengthening of the mitotic period rather than increased proliferation.


1975 ◽  
Vol 67 (3) ◽  
pp. 901-904 ◽  
Author(s):  
F Jauker

Protein and RNA contents of individual cells were measured cytophotometrically and related to the duration of individual generation times. Constant amounts of RNA per cell at division, and generation time-dependent protein contents, resulted in generation time-specific RNA/protein ratios. Experimental reduction of these ratios by inhibition of RNA synthesis stimulated premature macronuclear S phases.


1983 ◽  
Vol 50 (3) ◽  
pp. 357-363 ◽  
Author(s):  
Francisco J. Chavarri ◽  
Jose A. Nuñez ◽  
Manuel Nuñez

SummaryGeneration times and acid production after 6 and 24 h by 20 strains ofStreptococcus lactisof dairy origin were determined in heat treated (80 °C for 30 min) and sterilized cow's and ewe's milk. Ewe's milk enhanced growth of the streptococci, with significantly (P< 0·001) shorter generation times and higher acid production after 6 h incubation than cow's milk, probably due to its higher vitamin content. The stronger buffer capacity of ewe's milk allowed a higher (P< 0·001) acid production after 24 h than cow's milk. A stimulatory effect of sterilization on generation time and acid production after 24 h was observed in cow's milk. However, the heat treated ewe's milk was shown to be a better substrate than sterilized ewe's milk forStr. lactis.


2017 ◽  
Vol 10 (2) ◽  
pp. 156-161 ◽  
Author(s):  
Sophia F Shakur ◽  
Denise Brunozzi ◽  
Ahmed E Hussein ◽  
Andreas Linninger ◽  
Chih-Yang Hsu ◽  
...  

BackgroundThe hemodynamic evaluation of cerebral arteriovenous malformations (AVMs) using DSA has not been validated against true flow measurements.ObjectiveTo validate AVM hemodynamics assessed by DSA using quantitative magnetic resonance angiography (QMRA).Materials and methodsPatients seen at our institution between 2007 and 2016 with a supratentorial AVM and DSA and QMRA obtained before any treatment were retrospectively reviewed. DSA assessment of AVM flow comprised AVM arterial-to-venous time (A-Vt) and iFlow transit time. A-Vt was defined as the difference between peak contrast intensity in the cavernous internal carotid artery and peak contrast intensity in the draining vein. iFlow transit times were determined using syngo iFlow software. A-Vt and iFlow transit times were correlated with total AVM flow measured using QMRA and AVM angioarchitectural and clinical features.Results33 patients (mean age 33 years) were included. Nine patients presented with hemorrhage. Mean AVM volume was 9.8 mL (range 0.3–57.7 mL). Both A-Vt (r=−0.47, p=0.01) and iFlow (r=−0.44, p=0.01) correlated significantly with total AVM flow. iFlow transit time was significantly shorter in patients who presented with seizure but A-Vt and iFlow did not vary with other AVM angioarchitectural features such as venous stenosis or hemorrhagic presentation.ConclusionsA-Vt and iFlow transit times on DSA correlate with cerebral AVM flow measured using QMRA. Thus, these parameters may be used to indirectly estimate AVM flow before and after embolization during angiography in real time.


2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Nathan Moore ◽  
Stephen Lyle

Long-lived cancer stem cells (CSCs) with indefinite proliferative potential have been identified in multiple epithelial cancer types. These cells are likely derived from transformed adult stem cells and are thought to share many characteristics with their parental population, including a quiescent slow-cycling phenotype. Various label-retaining techniques have been used to identify normal slow cycling adult stem cell populations and offer a unique methodology to functionally identify and isolate cancer stem cells. The quiescent nature of CSCs represents an inherent mechanism that at least partially explains chemotherapy resistance and recurrence in posttherapy cancer patients. Isolating and understanding the cell cycle regulatory mechanisms of quiescent cancer cells will be a key component to creation of future therapies that better target CSCs and totally eradicate tumors. Here we review the evidence for quiescent CSC populations and explore potential cell cycle regulators that may serve as future targets for elimination of these cells.


Development ◽  
1993 ◽  
Vol 118 (2) ◽  
pp. 389-399 ◽  
Author(s):  
E.J. Sanders ◽  
M. Varedi ◽  
A.S. French

Cell proliferation in the gastrulating chick embryo was assessed using two independent techniques which mark cells in S phase of the mitotic cycle: nuclear incorporation of bromodeoxyuridine (BrdU) detected immunocytochemically and immunolocalization of proliferating cell nuclear antigen (PCNA). Computer-reconstructed maps were produced showing the distribution of labelled nuclei in the primitive streak and the cell layers. These distributions were also normalized to take into account regional differences in cell density across the embryo. Results from a 2 hour pulse of BrdU indicated that although cells at caudal levels of the primitive streak showed the highest incorporation, this region showed a similar proportion of labelled cells to the surrounding caudal regions of the epiblast and mesoderm when normalized for cell density. The entire caudal third of the embryo showed the highest proportion of cells in S phase. Cells of Hensen's node showed a relatively low rate of incorporation and, although the chordamesoderm cells showed many labelled nuclei, this appeared to be a reflection of a high cell density in this region. Combining this result with results from a 4 hour pulse of BrdU permitted mapping of cell generation time across the entire embryo. Generation times ranged from a low value of approximately 2 hours at caudal levels of both the epiblast and mesoderm, to an upper value of approximately 10 hours in the rostral regions of the primitive streak, in the mid-lateral levels of the epiblast and in the chordamesoderm rostral to Hensen's node. Cells at caudal regions of the primitive streak showed a generation time of approximately 5 hours. Taking into account that cells are generally considered to be continuously moving through the primitive streak, we conclude that cell division, as judged by generation time, is greatly reduced during transit through this region, despite the presence there of cells in S phase and M phase. Immunocytochemical localization of PCNA-positive nuclei gave generally similar distributions to those obtained with BrdU incorporation, confirming that this endogenous molecule is a useful S-phase marker during early embryogenesis. Mid-levels and caudal levels of the primitive streak showed the highest numbers of positive nuclei, and the highest proportion of labelling after cell density was accounted for. As with BrdU incorporation, the highest proportions of PCNA-positive nuclei were found towards the caudal regions of the epiblast and mesoderm. These results suggest that the differential growth of the caudal region of the embryo at this time is a direct consequence of elevated levels of cell proliferation in this region.(ABSTRACT TRUNCATED AT 400 WORDS)


2000 ◽  
Vol 2 (3) ◽  
pp. 215-232 ◽  
Author(s):  
Shea N. Gardner

Dose response curves show that prolonged drug exposure at a low concentration may kill more cells than short exposures at higher drug concentrations, particularly for cell cycle phase specific drugs. Applying drugs at low concentrations for prolonged periods, however, allows cells with partial resistance to evolve higher levels of resistance through stepwise processes such as gene amplification. Models are developed for cell cycle specific (CS) and cell cycle nonspecific (CNS) drugs to identify the schedule of drug application that balances this tradeoff.The models predict that a CS drug may be applied most effectively by splitting the cumulative dose into many (>40) fractions applied by long-term chemotherapy, while CNS drugs may be better applied in fewer than 10 fractions applied over a shorter term. The model suggests that administering each fraction by continuous infusion may be more effective than giving the drug as a bolus, whether the drug is CS or CNS. In addition, tumors with a low growth fraction or slow rate of cell division are predicted to be controlled more easily with CNS drugs, while those with a high proliferative fraction or fast cell division rate may respond better to CS drugs.


2009 ◽  
Vol 5 (3-4) ◽  
pp. 57-67 ◽  
Author(s):  
Konstantina Psachoulia ◽  
Francoise Jamen ◽  
Kaylene M. Young ◽  
William D. Richardson

Oligodendrocyte precursors (OLPs or ‘NG2 cells’) are abundant in the adult mouse brain, where they continue to proliferate and generate new myelinating oligodendrocytes. By cumulative BrdU labelling, we estimated the cell cycle timeTCand the proportion of NG2 cells that is actively cycling (the growth fraction) at ~ postnatal day 6 (P6), P60, P240 and P540. In the corpus callosum,TCincreased from <2 days at P6 to ~9 days at P60 to ~70 days at P240 and P540. In the cortex,TCincreased from ~2 days to >150 days over the same period. The growth fraction remained relatively invariant at ~50% in both cortex and corpus callosum – that is, similar numbers of mitotically active and inactive NG2 cells co-exist at all ages. Our data imply that a stable population of quiescent NG2 cells appears before the end of the first postnatal week and persists throughout life. The mitotically active population acts as a source of new oligodendrocytes during adulthood, while the biological significance of the quiescent population remains to be determined. We found that the mitotic status of adult NG2 cells is unrelated to their developmental site of origin in the ventral or dorsal telencephalon. We also report that new oligodendrocytes continue to be formed at a slow rate from NG2 cells even after P240 (8 months of age).


Many components of cell and nuclear size and mass are correlated with nuclear DNA content in plants, as also are the durations and rates of such developmental processes as mitosis and meiosis. It is suggested that the multiple effects of the mass of nuclear DNA which affect all cells and apply throughout the life of the plant can together determine the minimum generation time for each species. The durations of mitosis and of meiosis are both positively correlated with nuclear DNA content and, therefore, species with a short minimum generation time might be expected to have a shorter mean cell cycle time and mean meiotic duration, and a lower mean nuclear DNA content, than species with a long mean minimum generation time. In tests of this hypothesis, using data collated from the literature, it is shown that the mean cell cycle time and the mean meiotic duration in annual species is significantly shorter than in perennial species. Furthermore, the mean nuclear DNA content of annual species is significantly lower than for perennial species both in dicotyledons and monocotyledons. Ephemeral species have a significantly lower mean nuclear DNA content than annual species. Among perennial monocotyledons the mean nuclear DNA content of species which can complete a life cycle within one year (facultative perennials) is significantly lower than the mean nuclear DNA content of those which cannot (obligate perennials). However, the mean nuclear DNA content of facultative perennials does not differ significantly from the mean for annual species. It is suggested that the effects of nuclear DNA content on the duration of developmental processes are most obvious during its determinant stages, and that the largest effects of nuclear DNA mass are expressed at times when development is slowest, for instance, during meiosis or at low temperature. It has been suggested that DNA influences development in two ways, directly through its informational content, and indirectly by the physical-mechanical effects of its mass. The term 'nucleotype' is used to describe those conditions of the nucleus which effect the phenotype independently of the informational content of the DNA. It is suggested that cell cycle time, meiotic duration, and minimum generation time are determined by the nucleotype. In addition, it may be that satellite DNA is significant in its nucleotypic effects on developmental processes.


2020 ◽  
Author(s):  
Hirofumi Kiyokawa ◽  
Akira Yamaoka ◽  
Chisa Matsuoka ◽  
Tomoko Tokuhara ◽  
Takaya Abe ◽  
...  

SummaryDuring development, quiescent basal stem cells are derived from proliferative primordial progenitors through the cell cycle slowdown. In contrast, quiescent basal cells contribute to tissue repair during adult tissue regeneration by shifting from slow-cycling to proliferating and subsequently back to slow-cycling. Although sustained basal cell proliferation results in tumorigenesis, the molecular mechanisms regulating these transitions remain unknown. Using temporal single-cell transcriptomics of developing murine airway progenitors and in vivo genetic validation experiments, we found that Tgfß signaling slowed down cell cycle by inhibiting Id2 expression in airway progenitors and contributed to the specification of slow-cycling basal cell population during development. In adult tissue regeneration, reduced Tgfß signaling restored Id2 expression and initiated epithelial regeneration. Id2 overexpression and Tgfbr2 knockout enhanced epithelial proliferation; however, persistent Id2 expression in basal cells drove hyperplasia at a rate that resembled a precancerous state. Together, the Tgfß-Id2 axis commonly regulates the proliferation transitions in airway basal cells during development and regeneration, and its fine-tuning is critical for normal regeneration while avoiding basal cell hyperplasia.


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