Calcium and bicarbonate signaling pathways have pivotal, resonating roles in matching ATP production to demand
- Center for Biomedical Engineering and Technology, University of Maryland School of Medicine, United States
- Department of Physiology, University of Marylan School of Medicine, United States
- Claude D. Pepper Older Americans Independence Center, University of Maryland School of Medicine, United States
- Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, United States
- Marlene and Stewart Greenebaum Comprehensive Cancer Center, University of Maryland Baltimore School of Medicine, United States
- Division of Cardiovascular Medicine, Department of Medicine, University of Maryland School of Medicine, United States
Figures
Figure 1 with 1 supplement
Mitochondrial localization of soluble adenylyl cyclase (sAC).
(A) Airyscan super-resolution fluorescence images of a cardiomyocyte (left to right) immuno-labeled for sAC (yellow) and ATP synthase (red), and loaded with Alexa Fluor 488 phalloidin (1 M) to label …
-
Figure 1—source data 1
Numeric source data for Figure 1.
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Mitochondrial localization of bicarbonate-activated soluble adenylyl cyclase (sAC).
(A) Airyscan ‘super-resolution’ images of isolated cardiac mitochondria immuno-labeled for sAC (yellow) and ATP synthase (red). (B) Immunofluorescence profile of sAC (yellow) and ATP synthase (red) …
Figure 2 with 2 supplements
Mitochondrial function of soluble adenylyl cyclase (sAC).
(A) Quantitative ELISA measurements of cAMP inside isolated mitochondria (pmol/mg). cAMP is measured following treatment with indicated concentrations of bicarbonate (HCO3−), which activates sAC, …
-
Figure 2—source data 1
Numeric source data for Figure 2.
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Intra-mitochondrial localization of carbonic anhydrase 14 (CAXIV).
(A) Confocal images of isolated cardiac mitochondria immuno-labeled for either CA-II (left), or CA-Vb (middle), or CA-XIV (right). While three different primary antibodies were used to immuno-label …
Figure 2—video 1
Electron tomogram of a mitochondrion in the interfibrillar region of a rat cardiomyocyte.
Video is a slice-by-slice view of a subvolume of cardiac muscle (1840 × 1840 × 250 nm3) reconstructed by electron tomography.
Figure 3
cAMP control of mitochondrial ATP production.
(A) Left, sensitivity of mitochondrial ATP production to cAMP and phosphodiesterase inhibition (with 50 μM IBMX). Measurements are carried out at low [Ca2+]m (<200 nM). Right, sensitivity of …
-
Figure 3—source data 1
Western blot analysis in Figure 3C, D (anti-Rap1).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig3-data1-v2.zip
-
Figure 3—source data 2
Original file No. 1 for western blot analysis in Figure 3C, D (anti-Rap1).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig3-data2-v2.zip
-
Figure 3—source data 3
Original file No. 2 for western blot analysis in Figure 3C, D (anti-Rap1).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig3-data3-v2.zip
-
Figure 3—source data 4
Original file No. 3 for western blot analysis in Figure 3C, D (anti-Rap1).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig3-data4-v2.zip
-
Figure 3—source data 5
Original file No. 4 for western blot analysis in Figure 3C, D (anti-Rap1).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig3-data5-v2.zip
Figure 4
Elevated matrix [cAMP] inhibits mitochondrial ATP production.
(A) Diagram showing mitochondrial compartments accessible to cAMP and the membrane permeant analog, 8Br-cAMP. (B) Left, mitochondrial ATP production was decreased by application of 8Br-cAMP. This …
-
Figure 4—source data 1
Numeric source data for Figure 4.
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig4-data1-v2.xlsx
Figure 5
Dynamic control of mitochondrial ATP production by Ca2+ and bicarbonate.
Schematic diagram of the two complementary signaling arms that control mitochondrial ATP production. Left: The Ca2+ arm depends on heart rate. With higher heart rates, increased mitochondrial matrix …
Figure 6 with 2 supplements
Regulation of mitochondrial ATP production in the post myocardial infarction (MI) heart by [Ca2+]m and [cAMP].
(A) Anterior view of rat heart 8 weeks after induction of MI by ligation of left anterior descending (LAD) coronary artery. The black arrow indicates the location of the ligation and the distal, …
-
Figure 6—source data 1
Numeric source data for Figure 6.
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Regional contractile failure detected with speckle-tracking based strain analysis.
Time-resolved tracking of myocardial deformation for direct tracking of myocardial strain. (A) Top panel shows measurements carried out along the circumferential axis 8 weeks post-MI. Six regional …
Figure 6—figure supplement 2
Central components of mitochondrial ATP production are largely unchanged post-MI.
(A) Western blot analysis of mitochondria from the posterior wall 8 weeks after surgery (Post-MI or Sham). Tom20 is used as loading control. CV (ATP synthase subunit 5 A), CIV (complex 4 MTCO1), CII …
-
Figure 6—figure supplement 2—source data 1
Western blot analysis in Figure 3 (anti-Cv, anti-Civ, anti-Cii, anti-Ci, and anti-TOM20).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig6-figsupp2-data1-v2.zip
-
Figure 6—figure supplement 2—source data 2
Original file for western blot analysis of posterior wall in Figure 3 (anti-Cv, anti-Civ, anti-Cii, and anti-Ci).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig6-figsupp2-data2-v2.zip
-
Figure 6—figure supplement 2—source data 3
Original file for western blot analysis of posterior wall in Figure 3 (anti-TOM20).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig6-figsupp2-data3-v2.zip
-
Figure 6—figure supplement 2—source data 4
Original file for western blot analysis of septal wall in Figure 3 (anti-Cv, anti-Civ, anti-Cii, and anti-Ci).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig6-figsupp2-data4-v2.zip
-
Figure 6—figure supplement 2—source data 5
Original file for western blot analysis of septal wall in Figure 3 (anti-TOM20).
- https://cdn.elifesciences.org/articles/84204/elife-84204-fig6-figsupp2-data5-v2.zip
Additional files
-
Supplementary file 1
Mechanistic findings by investigations of sAC and its role in mitochondria.
- https://cdn.elifesciences.org/articles/84204/elife-84204-supp1-v2.docx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/84204/elife-84204-mdarchecklist1-v2.docx
