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Ammonia Affects Astroglial Proliferation in Culture.

Bodega G, Segura B, Ciordia S, Mena Mdel C, López-Fernández LA, García MI, Trabado I, Suárez I - PLoS ONE (2015)

Bottom Line: The DNA content was reduced in the treated cells, and BrdU incorporation diminished.A reduction in cell viability was also noted.The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biomedicina y Biotecnología, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain.

ABSTRACT
Primary cultures of rat astroglial cells were exposed to 1, 3 and 5 mM NH4Cl for up to 10 days. Dose- and time-dependent reductions in cell numbers were seen, plus an increase in the proportion of cells in the S phase. The DNA content was reduced in the treated cells, and BrdU incorporation diminished. However, neither ammonia nor ammonia plus glutamine had any effect on DNA polymerase activity. iTRAQ analysis showed that exposure to ammonia induced a significant reduction in histone and heterochromatin protein 1 expression. A reduction in cell viability was also noted. The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis.

No MeSH data available.


Related in: MedlinePlus

Effect of ammonia on cell cycle phases.Cell cycle analysis after 10 days of exposure to 1, 3 and 5 mM ammonia. The dotted lines are included to indicate the trends for the cell cycle phases. Columns and bars represent means and SDs. The percentages of cells in the phases considered (G0-G1, S and G2-M) are included for every experimental condition; it should be noted that the sum of the three phases is 100% (104 cells).
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pone.0139619.g004: Effect of ammonia on cell cycle phases.Cell cycle analysis after 10 days of exposure to 1, 3 and 5 mM ammonia. The dotted lines are included to indicate the trends for the cell cycle phases. Columns and bars represent means and SDs. The percentages of cells in the phases considered (G0-G1, S and G2-M) are included for every experimental condition; it should be noted that the sum of the three phases is 100% (104 cells).

Mentions: No significant differences were observed between treatments in terms of the number of cells in the G2-M phase. However, a clear increase in the number of cells in the S phase was detected (reaching 9% with the 5 mM ammonia treatment), with a concomitant reduction in the number of cells in the G0-G1 phase (Fig 4). Using the algorithm N = No x 2t/g (where N and No are the final and initial cell numbers, t the duration of treatment, and g the duration of the cell cycle), and taking into account (1) a cell cycle length 20 h [12], (2) 10 days of exposure, and (3) that ammonia induces a 10% time delay in the completion of the cell cycle, a theoretical reduction of 46% is obtained. This figure lies between the 40% and 50% reductions detected in cell count analysis.


Ammonia Affects Astroglial Proliferation in Culture.

Bodega G, Segura B, Ciordia S, Mena Mdel C, López-Fernández LA, García MI, Trabado I, Suárez I - PLoS ONE (2015)

Effect of ammonia on cell cycle phases.Cell cycle analysis after 10 days of exposure to 1, 3 and 5 mM ammonia. The dotted lines are included to indicate the trends for the cell cycle phases. Columns and bars represent means and SDs. The percentages of cells in the phases considered (G0-G1, S and G2-M) are included for every experimental condition; it should be noted that the sum of the three phases is 100% (104 cells).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4589356&req=5

pone.0139619.g004: Effect of ammonia on cell cycle phases.Cell cycle analysis after 10 days of exposure to 1, 3 and 5 mM ammonia. The dotted lines are included to indicate the trends for the cell cycle phases. Columns and bars represent means and SDs. The percentages of cells in the phases considered (G0-G1, S and G2-M) are included for every experimental condition; it should be noted that the sum of the three phases is 100% (104 cells).
Mentions: No significant differences were observed between treatments in terms of the number of cells in the G2-M phase. However, a clear increase in the number of cells in the S phase was detected (reaching 9% with the 5 mM ammonia treatment), with a concomitant reduction in the number of cells in the G0-G1 phase (Fig 4). Using the algorithm N = No x 2t/g (where N and No are the final and initial cell numbers, t the duration of treatment, and g the duration of the cell cycle), and taking into account (1) a cell cycle length 20 h [12], (2) 10 days of exposure, and (3) that ammonia induces a 10% time delay in the completion of the cell cycle, a theoretical reduction of 46% is obtained. This figure lies between the 40% and 50% reductions detected in cell count analysis.

Bottom Line: The DNA content was reduced in the treated cells, and BrdU incorporation diminished.A reduction in cell viability was also noted.The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biomedicina y Biotecnología, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain.

ABSTRACT
Primary cultures of rat astroglial cells were exposed to 1, 3 and 5 mM NH4Cl for up to 10 days. Dose- and time-dependent reductions in cell numbers were seen, plus an increase in the proportion of cells in the S phase. The DNA content was reduced in the treated cells, and BrdU incorporation diminished. However, neither ammonia nor ammonia plus glutamine had any effect on DNA polymerase activity. iTRAQ analysis showed that exposure to ammonia induced a significant reduction in histone and heterochromatin protein 1 expression. A reduction in cell viability was also noted. The ammonia-induced reduction of proliferative activity in these cultured astroglial cells seems to be due to a delay in the completion of the S phase provoked by the inhibition of chromatin protein synthesis.

No MeSH data available.


Related in: MedlinePlus