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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 liver cryostat sections. (A) Overview of PAG activity in mouse liver tissue in the presence of 30 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. Boxes show the measured (dotted line) and enlarged (continues line) areas in the tissue sections. Scale (B–H), 200 µm.
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fig10-0022155414551177: Phosphate-activated glutaminase (PAG) activity staining in mouse liver cryostat sections. (A) Overview of PAG activity in mouse liver tissue in the presence of 30 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. Boxes show the measured (dotted line) and enlarged (continues line) areas in the tissue sections. Scale (B–H), 200 µm.

Mentions: In the liver, PAG activity in the presence of low glutamine concentrations was highest in periportal areas as compared to pericentral areas after correcting for nonspecific background staining (e.g., staining in the absence of glutamine, Fig. 10). In the presence of 2 and 5 mM glutamine, PAG activity was approximately 1.5-fold higher in the periportal areas as compared with the pericentral areas. With glutamine concentrations higher than 5 mM, PAG activity zonation did not occur.


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 liver cryostat sections. (A) Overview of PAG activity in mouse liver tissue in the presence of 30 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. Boxes show the measured (dotted line) and enlarged (continues line) areas 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

fig10-0022155414551177: Phosphate-activated glutaminase (PAG) activity staining in mouse liver cryostat sections. (A) Overview of PAG activity in mouse liver tissue in the presence of 30 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. Boxes show the measured (dotted line) and enlarged (continues line) areas in the tissue sections. Scale (B–H), 200 µm.
Mentions: In the liver, PAG activity in the presence of low glutamine concentrations was highest in periportal areas as compared to pericentral areas after correcting for nonspecific background staining (e.g., staining in the absence of glutamine, Fig. 10). In the presence of 2 and 5 mM glutamine, PAG activity was approximately 1.5-fold higher in the periportal areas as compared with the pericentral areas. With glutamine concentrations higher than 5 mM, PAG activity zonation did not occur.

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