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Determination of phosphate-activated glutaminase activity and its kinetics in mouse tissues using metabolic mapping (quantitative enzyme histochemistry).

Botman D, Tigchelaar W, Van Noorden CJ - J. Histochem. Cytochem. (2014)

Bottom Line: PAG activity was decreased to 22% in the presence of 2 mM 6-diazo-5-oxo-L-norleucine.When compared with liver, kidney and brain, other tissues showed 3-fold to 6-fold less PAG activity.In conclusion, PAG is mainly active in mouse kidney, brain and liver, and shows different kinetics depending on which type of PAG is expressed.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).

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Phosphate-activated glutaminase (PAG) activity staining in mouse brain cryostat sections. (A) Overview of PAG activity in mouse brain tissue in the presence of 5 mM glutamine. Scale, 0.5 mm. PAG activity was stained with glutamate dehydrogenase (GDH) underneath the tissue sections in the presence of (B) 0 mM, (C) 2 mM, (D) 5 mM, (E) 10 mM, (F) 30 mM and (G) 50 mM glutamine and (H) without GDH underneath the tissue section. Box shows the enlarged area in the tissue sections. Scale (B–H), 200 µm.
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fig9-0022155414551177: Phosphate-activated glutaminase (PAG) activity staining in mouse brain cryostat sections. (A) Overview of PAG activity in mouse brain tissue in the presence of 5 mM glutamine. Scale, 0.5 mm. PAG activity was stained with glutamate dehydrogenase (GDH) underneath the tissue sections in the presence of (B) 0 mM, (C) 2 mM, (D) 5 mM, (E) 10 mM, (F) 30 mM and (G) 50 mM glutamine and (H) without GDH underneath the tissue section. Box shows the enlarged area in the tissue sections. Scale (B–H), 200 µm.

Mentions: In the brain, PAG showed a high and evenly distributed activity in the cortex (Fig. 9) and a low activity in myelin-rich structures (e.g., the corpus callosum).


Determination of phosphate-activated glutaminase activity and its kinetics in mouse tissues using metabolic mapping (quantitative enzyme histochemistry).

Botman D, Tigchelaar W, Van Noorden CJ - J. Histochem. Cytochem. (2014)

Phosphate-activated glutaminase (PAG) activity staining in mouse brain cryostat sections. (A) Overview of PAG activity in mouse brain tissue in the presence of 5 mM glutamine. Scale, 0.5 mm. PAG activity was stained with glutamate dehydrogenase (GDH) underneath the tissue sections in the presence of (B) 0 mM, (C) 2 mM, (D) 5 mM, (E) 10 mM, (F) 30 mM and (G) 50 mM glutamine and (H) without GDH underneath the tissue section. Box shows the enlarged area in the tissue sections. Scale (B–H), 200 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4230542&req=5

fig9-0022155414551177: Phosphate-activated glutaminase (PAG) activity staining in mouse brain cryostat sections. (A) Overview of PAG activity in mouse brain tissue in the presence of 5 mM glutamine. Scale, 0.5 mm. PAG activity was stained with glutamate dehydrogenase (GDH) underneath the tissue sections in the presence of (B) 0 mM, (C) 2 mM, (D) 5 mM, (E) 10 mM, (F) 30 mM and (G) 50 mM glutamine and (H) without GDH underneath the tissue section. Box shows the enlarged area in the tissue sections. Scale (B–H), 200 µm.
Mentions: In the brain, PAG showed a high and evenly distributed activity in the cortex (Fig. 9) and a low activity in myelin-rich structures (e.g., the corpus callosum).

Bottom Line: PAG activity was decreased to 22% in the presence of 2 mM 6-diazo-5-oxo-L-norleucine.When compared with liver, kidney and brain, other tissues showed 3-fold to 6-fold less PAG activity.In conclusion, PAG is mainly active in mouse kidney, brain and liver, and shows different kinetics depending on which type of PAG is expressed.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands (DB, WT, CJFVN).

Show MeSH