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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 polymerase.Effect of ammonia (NH4Cl) on the activity of DNA polymerase, analyzed via real-time PCR. Note that the Ct variation ranges between 20 and 21. Columns and bars represent means and SDs. No significant changes were observed.
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pone.0139619.g007: Effect of ammonia on DNA polymerase.Effect of ammonia (NH4Cl) on the activity of DNA polymerase, analyzed via real-time PCR. Note that the Ct variation ranges between 20 and 21. Columns and bars represent means and SDs. No significant changes were observed.

Mentions: To determine whether the delay in DNA synthesis was due to an ammonia dysfunction involving DNA polymerase, real-time PCR analysis starting with the same quantity of cDNA was performed with or without ammonia in the reaction medium. Fig 7 shows that ammonia had no significant effect on DNA amplification or, consequently, on DNA polymerase function. Given that a difference of 1 unit in Ct corresponds to a 2x-fold expression change, the differences observed under the different ammonia conditions are extremely small.


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 polymerase.Effect of ammonia (NH4Cl) on the activity of DNA polymerase, analyzed via real-time PCR. Note that the Ct variation ranges between 20 and 21. Columns and bars represent means and SDs. No significant changes were observed.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139619.g007: Effect of ammonia on DNA polymerase.Effect of ammonia (NH4Cl) on the activity of DNA polymerase, analyzed via real-time PCR. Note that the Ct variation ranges between 20 and 21. Columns and bars represent means and SDs. No significant changes were observed.
Mentions: To determine whether the delay in DNA synthesis was due to an ammonia dysfunction involving DNA polymerase, real-time PCR analysis starting with the same quantity of cDNA was performed with or without ammonia in the reaction medium. Fig 7 shows that ammonia had no significant effect on DNA amplification or, consequently, on DNA polymerase function. Given that a difference of 1 unit in Ct corresponds to a 2x-fold expression change, the differences observed under the different ammonia conditions are extremely small.

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