Key resources
| Reagent or resource . | Source . | Identifier . |
|---|---|---|
| Antibodies | ||
| Guinea pig anti-vGLUT1 | Millipore Sigma | CAT#: AB5905 |
| Anti-Tuj1 (β-tubulin III) antibody | R&D Systems | MAB1195 |
| Anti-ATPase5β antibody | Sigma-Aldrich | CAT#: HPA001520 |
| Anti-Sirt3 antibody | Cell Signaling Technology | CAT#: 5490S |
| Anti-β actin antibody | Bio-Rad | CAT#: HCA147P |
| Anti-acetylated lysine antibody | Cell Signaling Technology | CAT#: 9441S |
| anti-α tubulin antibody | Sigma-Aldrich | CAT#: T6074 |
| Anti-TOMM20 antibody | Cell Signaling Technology | CAT#: 42406S |
| Anti-pCREB antibody | Cell Signaling Technology | CAT#: 9198S |
| Biological samples | ||
| rSirt3 shRNA lentivirus | Hope center viral vectors core at Washington University in St. Louis | N/A |
| PGC1α AAV | Gift of B. Finck | N/A |
| GFP AAV | Gift of B. Finck | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Dorsomorphin | Sigma-Aldrich | CAT#: 866405-64-3 |
| Deacetylation inhibition cocktail | Santa Cruz | CAT#: sc-362323 |
| 6-Cyano-7nitroquinoxalibe-2, 3-dione (CNQX) | Sigma-Aldrich | CAT#: C239-25 MG |
| D,L-2-amino-5phosphonovaleric acid (APV) | Sigma-Aldrich | CAT#: A5282 |
| Propidium iodide | Invitrogen | CAT#: P1304MP |
| Critical commercial assays | ||
| All-in-one 5X RT master mix | Applied Biosytems | CAT#: ABM-G592 |
| PowerUp SYBR green master mix | Thermo Fisher Scientific | CAT#: A25742 |
| TaqMan fast advanced master mix | Applied Biosytems, | CAT#: 4444557 |
| Deposited Data | ||
| RNA-seq data | Dryad repository | Tiwari et al., 2023 Preprint |
| Syn-ATP analysis code | GitHub | https://github.com/ashrafilab/SynATP-Analysis |
| Experimental models: Cell lines | ||
| Experimental models: Organisms/Strains | ||
| Sprague-Dawley rat | Charles River | Strain code: CD IGS SD # 001 RRID: RGD_734476 |
| C57BL/6NJ mice (overnight starvation) | Jackson Labs | Strain code: 005304 RRID:IMSR_JAX:005304 |
| C57BL/6J mice (alternate day fasting) | Jackson Labs | Strain code: 000664 RRID:IMSR_JAX:000664 |
| Recombinant DNA | ||
| vGLUT1-pHluorin | Voglmaier et al. (2006) | N/A |
| Syn-ATP | Rangaraju et al. (2014) | N/A |
| Synaptophysin-GCaMP6f | de Juan-Sanz et al. (2017) | N/A |
| mito4x-RFP | Gift of T.A. Ryan | N/A |
| mito4xi-GCaMP6f | Ashrafi et al. (2020) | #192075; Addgene |
| pLKO.1 vector | Moffat et al. (2006) | #10878; Addgene |
| hSirt3-RFP | This paper | |
| Sirt3-FLAG | North et al. (2003) | #13814; Addgene |
| Sequence-based reagents | ||
| rSirt3 shRNA (pLKO.1 vector) | This paper | Target sequence: 5′-CTTACTACATGTGGCTGAT-3′ |
| qPCR primers | Source | Sequence (5′-3′) |
| Mouse Pgc1α forward primer | Thermo Fisher Scientific | TGAAAAAGCTTGACTGGCGT |
| Mouse Pgc1α reverse primer | Thermo Fisher Scientific | CAACCAGAGCAGCACACTCTA |
| Rat Pgc1α forward primer | Thermo Fisher Scientific | TGAAAAAGCTTGACTGGCGT |
| Rat Pgc1α reverse primer | Thermo Fisher Scientific | CAGGGCAGCACACTCTATGT |
| Rat Sirt3 forward primer | Thermo Fisher Scientific | TGTGGGGTCCGGGAGTATTA |
| Rat Sirt3 reverse primer | Thermo Fisher Scientific | ACAGCTTCTTCTCACTGTGTCC |
| Rat Gapdh forward primer | Thermo Fisher Scientific | TGTGCAGTGCCAGCCTC |
| Rat Gapdh reverse primer | Thermo Fisher Scientific | AGAGAAGGCAGCCCTGGTAA |
| Rat 18 s Taqman probe | Thermo Fisher Scientific, #4331182 | N/A |
| Rat Sirt3 Taqman probe | Thermo Fisher Scientific, #4331182 | N/A |
| Rat β-actin forward primer | Thermo Fisher Scientific | GTCGAGTCCGCGTCCAC |
| Rat β-actin reverse primer | Thermo Fisher Scientific | TATCGTCATCCATGGCGAACTGG |
| Software and algorithms | ||
| ImageJ | National Institute of Health | https://imagej.nih.gov/ij/ |
| GraphPad prism 9.2.0 | GraphPad Software | http://www.graphpad.com/scientific-software/prism/ |
| Jupyter notebook | Project Jupyter | https://jupyter.org/ |
| Adobe illustrator | Adobe, Inc. | https://www.adobe.com |
| Reagent or resource . | Source . | Identifier . |
|---|---|---|
| Antibodies | ||
| Guinea pig anti-vGLUT1 | Millipore Sigma | CAT#: AB5905 |
| Anti-Tuj1 (β-tubulin III) antibody | R&D Systems | MAB1195 |
| Anti-ATPase5β antibody | Sigma-Aldrich | CAT#: HPA001520 |
| Anti-Sirt3 antibody | Cell Signaling Technology | CAT#: 5490S |
| Anti-β actin antibody | Bio-Rad | CAT#: HCA147P |
| Anti-acetylated lysine antibody | Cell Signaling Technology | CAT#: 9441S |
| anti-α tubulin antibody | Sigma-Aldrich | CAT#: T6074 |
| Anti-TOMM20 antibody | Cell Signaling Technology | CAT#: 42406S |
| Anti-pCREB antibody | Cell Signaling Technology | CAT#: 9198S |
| Biological samples | ||
| rSirt3 shRNA lentivirus | Hope center viral vectors core at Washington University in St. Louis | N/A |
| PGC1α AAV | Gift of B. Finck | N/A |
| GFP AAV | Gift of B. Finck | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Dorsomorphin | Sigma-Aldrich | CAT#: 866405-64-3 |
| Deacetylation inhibition cocktail | Santa Cruz | CAT#: sc-362323 |
| 6-Cyano-7nitroquinoxalibe-2, 3-dione (CNQX) | Sigma-Aldrich | CAT#: C239-25 MG |
| D,L-2-amino-5phosphonovaleric acid (APV) | Sigma-Aldrich | CAT#: A5282 |
| Propidium iodide | Invitrogen | CAT#: P1304MP |
| Critical commercial assays | ||
| All-in-one 5X RT master mix | Applied Biosytems | CAT#: ABM-G592 |
| PowerUp SYBR green master mix | Thermo Fisher Scientific | CAT#: A25742 |
| TaqMan fast advanced master mix | Applied Biosytems, | CAT#: 4444557 |
| Deposited Data | ||
| RNA-seq data | Dryad repository | Tiwari et al., 2023 Preprint |
| Syn-ATP analysis code | GitHub | https://github.com/ashrafilab/SynATP-Analysis |
| Experimental models: Cell lines | ||
| Experimental models: Organisms/Strains | ||
| Sprague-Dawley rat | Charles River | Strain code: CD IGS SD # 001 RRID: RGD_734476 |
| C57BL/6NJ mice (overnight starvation) | Jackson Labs | Strain code: 005304 RRID:IMSR_JAX:005304 |
| C57BL/6J mice (alternate day fasting) | Jackson Labs | Strain code: 000664 RRID:IMSR_JAX:000664 |
| Recombinant DNA | ||
| vGLUT1-pHluorin | Voglmaier et al. (2006) | N/A |
| Syn-ATP | Rangaraju et al. (2014) | N/A |
| Synaptophysin-GCaMP6f | de Juan-Sanz et al. (2017) | N/A |
| mito4x-RFP | Gift of T.A. Ryan | N/A |
| mito4xi-GCaMP6f | Ashrafi et al. (2020) | #192075; Addgene |
| pLKO.1 vector | Moffat et al. (2006) | #10878; Addgene |
| hSirt3-RFP | This paper | |
| Sirt3-FLAG | North et al. (2003) | #13814; Addgene |
| Sequence-based reagents | ||
| rSirt3 shRNA (pLKO.1 vector) | This paper | Target sequence: 5′-CTTACTACATGTGGCTGAT-3′ |
| qPCR primers | Source | Sequence (5′-3′) |
| Mouse Pgc1α forward primer | Thermo Fisher Scientific | TGAAAAAGCTTGACTGGCGT |
| Mouse Pgc1α reverse primer | Thermo Fisher Scientific | CAACCAGAGCAGCACACTCTA |
| Rat Pgc1α forward primer | Thermo Fisher Scientific | TGAAAAAGCTTGACTGGCGT |
| Rat Pgc1α reverse primer | Thermo Fisher Scientific | CAGGGCAGCACACTCTATGT |
| Rat Sirt3 forward primer | Thermo Fisher Scientific | TGTGGGGTCCGGGAGTATTA |
| Rat Sirt3 reverse primer | Thermo Fisher Scientific | ACAGCTTCTTCTCACTGTGTCC |
| Rat Gapdh forward primer | Thermo Fisher Scientific | TGTGCAGTGCCAGCCTC |
| Rat Gapdh reverse primer | Thermo Fisher Scientific | AGAGAAGGCAGCCCTGGTAA |
| Rat 18 s Taqman probe | Thermo Fisher Scientific, #4331182 | N/A |
| Rat Sirt3 Taqman probe | Thermo Fisher Scientific, #4331182 | N/A |
| Rat β-actin forward primer | Thermo Fisher Scientific | GTCGAGTCCGCGTCCAC |
| Rat β-actin reverse primer | Thermo Fisher Scientific | TATCGTCATCCATGGCGAACTGG |
| Software and algorithms | ||
| ImageJ | National Institute of Health | https://imagej.nih.gov/ij/ |
| GraphPad prism 9.2.0 | GraphPad Software | http://www.graphpad.com/scientific-software/prism/ |
| Jupyter notebook | Project Jupyter | https://jupyter.org/ |
| Adobe illustrator | Adobe, Inc. | https://www.adobe.com |