Organ-specific distribution of ACE2 mRNA and correlating peptidase activity in rodents

Florian Gembardt, Anja Sterner-Kock, Hans Imboden, Matthias Spalteholz, Franziska Reibitz, Heinz-Peter Schultheiss, Wolf-Eberhard Siems, Thomas Walther, Florian Gembardt, Anja Sterner-Kock, Hans Imboden, Matthias Spalteholz, Franziska Reibitz, Heinz-Peter Schultheiss, Wolf-Eberhard Siems, Thomas Walther

Abstract

Biochemical analysis revealed that angiotensin-converting enzyme related carboxy-peptidase (ACE2) cleaves angiotensin (Ang) II to Ang-(1-7), a heptapeptide identified as an endogenous ligand for the G protein-coupled receptor Mas. No data are currently available that systematically describe ACE2 distribution and activity in rodents. Therefore, we analyzed the ACE2 expression in different tissues of mice and rats on mRNA (RNase protection assay) and protein levels (immunohistochemistry, ACE2 activity, western blot). Although ACE2 mRNA in both investigated species showed the highest expression in the ileum, the mouse organ exceeded rat ACE2, as also demonstrated in the kidney and colon. Corresponding to mRNA, ACE2 activity was highest in the ileum and mouse kidney but weak in the rat kidney, which was also confirmed by immunohistochemistry. Contrary to mRNA, we found weak activity in the lung of both species. Our data demonstrate a tissue- and species-specific pattern for ACE2 under physiological conditions.

Figures

Fig. 1
Fig. 1
mRNA expression in different mouse tissues. Representative RPA of different tissues from C57Bl/6 mice. The specific bands for MMACE2 and the housekeeping mRNA rl32 are indicated with arrows on the left. The MMACE2 and rL32 probes are indicated with arrows on the right. (a) 1. ventricle, 2. kidney, 3. lung, 4. liver, 5. testis, 6. bladder, 7. forebrain, 8. spleen, y+, yeast plus RNase; y−, yeast without RNase; (b) 9. thymus, 2. kidney, 10. stomach, 11. ileum, 12. colon, 13. brainstem, 14. atrium, 15. adipose tissue; MM, Mus Musculus; y+, yeast plus RNase; y−, yeast without RNase.
Fig. 2
Fig. 2
mRNA expression of different rat tissues. Representative RPA of different tissues from C57Bl/6 mice. The specific bands for RNACE2 and the housekeeping mRNA rl32 are indicated with arrows on the left. The RNACE2 and rL32 probes are indicated with arrows on the right. (a) 1. ventricle, 2. kidney, 3. lung, 4. liver, 5. testis, 6. bladder, 7. forebrain, 8. spleen, y+, yeast plus RNase; y−, yeast without RNase; (b) 9. thymus, 2. kidney, 10. stomach, 11. ileum, 12. colon, 13. brainstem, 14. atrium, 15. adipose tissue; RN, Rattus Norvegicus; y+, yeast plus RNase; y−, yeast without RNase.
Fig. 3
Fig. 3
Quantification of the RPAs of mice (white columns) and rats (black columns). The mRNA amount of the lungs is set to 100% (n ≤ 4). The values are shown as mean + S.E.M. 1. ventricle, 2. kidney, 3. lung, 4. liver, 5. testis, 6. bladder, 7. forebrain, 8. spleen, 9. thymus, 10. stomach, 11. ileum, 12. colon, 13. brainstem, 14. atrium, 15. adipose tissue. *P < 0.05, **P < 0.01, ***P < 0.0001 compared mouse vs. rat.
Fig. 4
Fig. 4
Representative western blot of mouse (upper panel) and rat (lower panel) tissues with a commercial polyclonal antibody against ACE2. The band at 75 kDa is indicated. 1. atrium, 2. ventricle, 3. thymus, 4. spleen, 5. kidney, 6. testis, 7. lung, 8. bladder, 9. forebrain, 10. adipose tissue.
Fig. 5
Fig. 5
Immunohistochemical visualization of ACE2 positive cells. Sections of lungs (upper row) and kidneys (lower row) from mouse (left panel) and rat (right panel). In the lungs of both species alveolar macrophages and type 2 cells were stained positive. The tubulus epithelium in mouse kidney was stained positive, whereas in the rat kidney only weak staining was seen.

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