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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 kidney tissue. (A) Stitched overview image of PAG activity in mouse kidney cryostat sections in the presence of 30 mM glutamine. Scale, 1 mm. (B) Example of PAG activity in a glomerulus. Scale, 50 µm. (C) PAG activity staining in the presence of 50 mM glutamine without glutamate dehydrogenase (GDH) underneath the tissue section. PAG activity was stained with GDH underneath the tissue sections in the presence of (D) 0 mM, (E) 2 mM, (F) 5 mM, (G) 10 mM, (H) 30 mM and (I) 50 mM glutamine. Boxes show measured (dotted line) and enlarged (continues line) areas in the specific tissue sections. Scale (C–I), 200 µm. Image stitching was performed using Adobe Photoshop CS6 (Adobe, San Jose, CA).
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fig8-0022155414551177: Phosphate-activated glutaminase (PAG) activity staining in kidney tissue. (A) Stitched overview image of PAG activity in mouse kidney cryostat sections in the presence of 30 mM glutamine. Scale, 1 mm. (B) Example of PAG activity in a glomerulus. Scale, 50 µm. (C) PAG activity staining in the presence of 50 mM glutamine without glutamate dehydrogenase (GDH) underneath the tissue section. PAG activity was stained with GDH underneath the tissue sections in the presence of (D) 0 mM, (E) 2 mM, (F) 5 mM, (G) 10 mM, (H) 30 mM and (I) 50 mM glutamine. Boxes show measured (dotted line) and enlarged (continues line) areas in the specific tissue sections. Scale (C–I), 200 µm. Image stitching was performed using Adobe Photoshop CS6 (Adobe, San Jose, CA).

Mentions: In kidney, PAG activity was high in the cortex and inner medulla but low in the outer medulla, papillary tip and glomeruli (Fig. 8). The outer medulla showed regions with low activity that were most likely clusters of lis of Henle.


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 kidney tissue. (A) Stitched overview image of PAG activity in mouse kidney cryostat sections in the presence of 30 mM glutamine. Scale, 1 mm. (B) Example of PAG activity in a glomerulus. Scale, 50 µm. (C) PAG activity staining in the presence of 50 mM glutamine without glutamate dehydrogenase (GDH) underneath the tissue section. PAG activity was stained with GDH underneath the tissue sections in the presence of (D) 0 mM, (E) 2 mM, (F) 5 mM, (G) 10 mM, (H) 30 mM and (I) 50 mM glutamine. Boxes show measured (dotted line) and enlarged (continues line) areas in the specific tissue sections. Scale (C–I), 200 µm. Image stitching was performed using Adobe Photoshop CS6 (Adobe, San Jose, CA).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig8-0022155414551177: Phosphate-activated glutaminase (PAG) activity staining in kidney tissue. (A) Stitched overview image of PAG activity in mouse kidney cryostat sections in the presence of 30 mM glutamine. Scale, 1 mm. (B) Example of PAG activity in a glomerulus. Scale, 50 µm. (C) PAG activity staining in the presence of 50 mM glutamine without glutamate dehydrogenase (GDH) underneath the tissue section. PAG activity was stained with GDH underneath the tissue sections in the presence of (D) 0 mM, (E) 2 mM, (F) 5 mM, (G) 10 mM, (H) 30 mM and (I) 50 mM glutamine. Boxes show measured (dotted line) and enlarged (continues line) areas in the specific tissue sections. Scale (C–I), 200 µm. Image stitching was performed using Adobe Photoshop CS6 (Adobe, San Jose, CA).
Mentions: In kidney, PAG activity was high in the cortex and inner medulla but low in the outer medulla, papillary tip and glomeruli (Fig. 8). The outer medulla showed regions with low activity that were most likely clusters of lis of Henle.

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