Premium
Mitochondrial Transplantation in Cardiomyocytes: Rescue of Mitochondrial Function and Replacement of mtDNA
Author(s) -
Pacak Christina,
Preble Janine,
Kondo Hiroshi,
Levitsky Sidney,
Nido Pedro,
Cowan Douglas,
McCully James
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.764.4
Subject(s) - mitochondrial dna , transplantation , function (biology) , mitochondrion , biology , microbiology and biotechnology , medicine , genetics , gene
Background Previously, we have demonstrated that the transplantation of mitochondria into the myocardium significantly enhanced cardioprotection. Our studies established that mitochondria are internalized into cardiomyocytes following transplantation; however, the mechanism(s) modulating internalization of these organelles was unknown. Methods: Neonatal rat cardiomyocytes were cultured and pre‐treated with either cytochalasin D to block actin polymerization, methyl‐β‐cyclodextrin to block caveola‐dependent‐clathrin dependent endocytosis, nocodazole to block tunneling nano tubes or 5‐(N‐Ethyl‐N‐isopropyl)amiloride to block macro‐pinocytosis and then co‐incubated with isolated mitochondria pre‐labeled with a fluorescent marker (pHrodo). Mitochondrial internalization and ATP content was determined. The functional contribution of internalized mitochondria, was determined using HeLa p 0 cells depleted of mtDNA co‐incubated with mitochondria isolated from HeLa cells with intact mtDNA. Results: Internalization of mitochondria into cardiomyocytes was modulated by actin‐dependent endocytosis. Internalization of HeLa cell mitochondria into HeLa p 0 cells increased ATP content (12.74±4.6 vs 73.7±17.2 nmol ATP/ 10 3 cells; p<0.05) and oxygen consumption rate (0.55±0.21 vs. 2.32±0.94; p<0.05) and replaced depleted mtDNA. Conclusions These results provide a mechanism for the internalization of mitochondria within host cells and a basis for novel therapeutic interventions allowing for the rescue and replacement of damaged or impaired mitochondria.