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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 BrdU incorporation.(A) BrdU incorporation by astroglial cells following exposure to ammonia. Points and bars represent means and SD. No significant changes were observed. (B) Cultured cells showing BrdU incorporation (red): (1) 1 day control, (2) 3 days after exposure to 5 mM ammonia. Scale bar: 10 μm.
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pone.0139619.g006: Effect of ammonia on BrdU incorporation.(A) BrdU incorporation by astroglial cells following exposure to ammonia. Points and bars represent means and SD. No significant changes were observed. (B) Cultured cells showing BrdU incorporation (red): (1) 1 day control, (2) 3 days after exposure to 5 mM ammonia. Scale bar: 10 μm.

Mentions: The delay in the completion of the S phase was examined by BrdU incorporation. BrdU is incorporated into newly synthesized DNA in the place of thymidine, and is very easy to detect; slower DNA synthesis should thus be revealed by reduced BrdU incorporation. BrdU was incorporated to a lesser extent by ammonia-treated than by control cells (Fig 6). At 10 days, the 5 mM ammonia-treated cells incorporated 45.5% less BrdU than did controls, in line with the reduction recorded 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 BrdU incorporation.(A) BrdU incorporation by astroglial cells following exposure to ammonia. Points and bars represent means and SD. No significant changes were observed. (B) Cultured cells showing BrdU incorporation (red): (1) 1 day control, (2) 3 days after exposure to 5 mM ammonia. Scale bar: 10 μm.
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Related In: Results  -  Collection

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

pone.0139619.g006: Effect of ammonia on BrdU incorporation.(A) BrdU incorporation by astroglial cells following exposure to ammonia. Points and bars represent means and SD. No significant changes were observed. (B) Cultured cells showing BrdU incorporation (red): (1) 1 day control, (2) 3 days after exposure to 5 mM ammonia. Scale bar: 10 μm.
Mentions: The delay in the completion of the S phase was examined by BrdU incorporation. BrdU is incorporated into newly synthesized DNA in the place of thymidine, and is very easy to detect; slower DNA synthesis should thus be revealed by reduced BrdU incorporation. BrdU was incorporated to a lesser extent by ammonia-treated than by control cells (Fig 6). At 10 days, the 5 mM ammonia-treated cells incorporated 45.5% less BrdU than did controls, in line with the reduction recorded 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