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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 DNA content.(A) Four rows of the in-cell DNA content analysis plate; note that ammonia exposure reduced the fluorescence signal. The row shows only nine results since the external wells were not seeded and the control point without fluorescent dye is not included. (B) Plot showing the change in well DNA content after ammonia exposure. Points and bars represent means and SDs. **p<0.05, ***p<0.01.
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pone.0139619.g005: Effect of ammonia on DNA content.(A) Four rows of the in-cell DNA content analysis plate; note that ammonia exposure reduced the fluorescence signal. The row shows only nine results since the external wells were not seeded and the control point without fluorescent dye is not included. (B) Plot showing the change in well DNA content after ammonia exposure. Points and bars represent means and SDs. **p<0.05, ***p<0.01.

Mentions: A lower cell proliferation rate leads to a reduction in cell number, which should be associated with a lower well DNA content. This was measured using an in-cell based assay to avoid protocols including cell fragmentation and DNA extraction. In general, the well DNA content was reduced when cells were treated with ammonia, with significant changes observed after 5 days of exposure to the 5 mM concentration, and after 10 days of exposure to the 3 mM concentration (Fig 5).


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 DNA content.(A) Four rows of the in-cell DNA content analysis plate; note that ammonia exposure reduced the fluorescence signal. The row shows only nine results since the external wells were not seeded and the control point without fluorescent dye is not included. (B) Plot showing the change in well DNA content after ammonia exposure. Points and bars represent means and SDs. **p<0.05, ***p<0.01.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139619.g005: Effect of ammonia on DNA content.(A) Four rows of the in-cell DNA content analysis plate; note that ammonia exposure reduced the fluorescence signal. The row shows only nine results since the external wells were not seeded and the control point without fluorescent dye is not included. (B) Plot showing the change in well DNA content after ammonia exposure. Points and bars represent means and SDs. **p<0.05, ***p<0.01.
Mentions: A lower cell proliferation rate leads to a reduction in cell number, which should be associated with a lower well DNA content. This was measured using an in-cell based assay to avoid protocols including cell fragmentation and DNA extraction. In general, the well DNA content was reduced when cells were treated with ammonia, with significant changes observed after 5 days of exposure to the 5 mM concentration, and after 10 days of exposure to the 3 mM concentration (Fig 5).

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