Abstract
Adenine nucleotide translocases (ANTs) transport ADP and ATP through mitochondrial inner membrane, thus playing an essential role for energy metabolism of eukaryotic cells. Mice have three ANT paralogs, Ant1 (Slc25a4), Ant2 (Slc25a5) and Ant4 (Slc25a31), which are expressed in a tissue-dependent manner. While knockout mice have been characterized with Ant1 and Ant4 genes, which resulted in exercise intolerance and male infertility, respectively, the role of the ubiquitously expressed Ant2 gene in animal development has not been fully demonstrated. Here, we generated Ant2 hypomorphic mice by targeted disruption of the gene, in which Ant2 expression is largely depleted. The mice showed apparently normal embryonic development except pale phenotype along with a reduced birth rate. However, postnatal growth was severely retarded with macrocytic anemia, B lymphocytopenia, lactic acidosis and bloated stomach, and died within 4 weeks. Ant2 depletion caused anemia in a cell-autonomous manner by maturation arrest of erythroid precursors with increased reactive oxygen species and premature deaths. B-lymphocyte development was similarly affected by Ant2 depletion, and splenocytes showed a reduction in maximal respiration capacity and cellular ATP levels as well as an increase in cell death accompanying mitochondrial permeability transition pore opening. In contrast, myeloid, megakaryocyte and T-lymphocyte lineages remained apparently intact. Erythroid and B-cell development may be particularly vulnerable to Ant2 depletion-mediated mitochondrial dysfunction and oxidative stress.
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Abbreviations
- 7AAD:
-
7-aminoactinomycin D
- ANT:
-
adenine nucleotide translocase
- BM:
-
bone marrow
- CFU-E:
-
colony-forming unit-erythroid
- CFU-GM:
-
colony-forming unit-granulocyte macrophage
- CsA:
-
cyclosporin A
- ECAR:
-
extracellular acidification rate
- ES cell:
-
embryonic stem cell
- FCCP:
-
fluoro-carbonyl cyanide phenylhydrazone
- FL:
-
fetal liver
- GI:
-
gastrointestinal
- HCT:
-
hematocrit
- HGB:
-
hemoglobin
- HSV-TK:
-
herpes simplex virus thymidine kinase
- IRES:
-
internal ribosome entry site
- LSK:
-
lineage-negative Sca1 and c-Kit positive
- MCV:
-
mean corpuscular volume
- MNGIE:
-
mitochondrial neurogastrointestinal encephalopathy
- mPTP:
-
mitochondrial permeability transition pore
- mtDNA:
-
mitochondrial deoxyribonucleic acid
- OCR:
-
oxygen consumption rate
- PARP:
-
poly ADP ribose polymerase
- PB:
-
peripheral blood
- PI:
-
propidium iodide
- POLG:
-
polymerase γ
- RBC:
-
red blood cell
- RDW:
-
red blood cell distribution width
- ROS:
-
reactive oxygen species
- SIBN:
-
splicing acceptor-IRES-β-galactosidase-neomycin
- SOD:
-
superoxide dismutase
- TMRM:
-
tetramethyl rhodamine methyl ester
- TP:
-
thymidine phosphorylase
- TUNEL:
-
terminal deoxynuclotidyl transferase-mediated dUTP nick-end labeling
- WT:
-
wild type
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Acknowledgements
We thank Dr. Laurence Morel (University of Florida) for providing antibodies for hematopoietic precursor sorting, Dr. Brian Harfe for the Flp vector, Drs. Clayton Matthews, William Dunn, Chen Liu and W Strat May for helpful discussion and technical advice and Dr. Katherine Santostefano for critical reading of the manuscript. We also appreciate Dr. Robert Anderson (Newcastle University, UK) for his advice on cardiac histology. This research was supported in part by National Institutes of Health Grants (HD060474 to NT; DK079879 and DK090115 to JK), and a research grant from Otsuka Maryland Medicinal Laboratories to NT.
Author Contributions
JC performed research, analyzed data, designed research and wrote the paper; JS, CHL, TS, M-HL, RRC performed research; YJL, HK, J-SK analyzed data; SPO analyzed data and designed research; YJL performed research, analyzed data and designed research; and NT designed research, analyzed data and wrote the paper.
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Cho, J., Seo, J., Lim, C. et al. Mitochondrial ATP transporter Ant2 depletion impairs erythropoiesis and B lymphopoiesis. Cell Death Differ 22, 1437–1450 (2015). https://doi.org/10.1038/cdd.2014.230
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DOI: https://doi.org/10.1038/cdd.2014.230
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