title: Differential Expression of Complement Markers in Normal and AMD Transmitochondrial Cybrids.
abstract: OBJECTIVE
  Variations in mitochondrial DNA (mtDNA) and abnormalities in the complement pathways have been implicated in the pathogenesis of age-related macular degeneration (AMD). This study was designed to determine the effects of mtDNA from AMD subjects on the complement pathway.
  
  
  METHODS
  Transmitochondrial cybrids were prepared by fusing platelets from AMD and age-matched Normal subjects with Rho0 (lacking mtDNA) human ARPE-19 cells. Quantitative PCR and Western blotting were performed to examine gene and protein expression profiles, respectively, of complement markers in these cybrids. Bioenergetic profiles of Normal and AMD cybrids were examined using the Seahorse XF24 flux analyzer.
  
  
  RESULTS
  Significant decreases in the gene and protein expression of complement inhibitors, along with significantly higher levels of complement activators, were found in AMD cybrids compared to Older-Normal cybrids. Seahorse flux data demonstrated that the bioenergetic profiles for Older-Normal and Older-AMD cybrid samples were similar to each other but were lower compared to Young-Normal cybrid samples.
  
  
  CONCLUSIONS
  In summary, since all cybrids had identical nuclei and differed only in mtDNA content, the observed changes in components of complement pathways can be attributed to mtDNA variations in the AMD subjects, suggesting that mitochondrial genome and retrograde signaling play critical roles in this disease. Furthermore, the similar bioenergetic profiles of AMD and Older-Normal cybrids indicate that the signaling between mitochondria and nuclei are probably not via a respiratory pathway.
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.
authors.forename: Sonali
authors.initials: S
authors.lastname: Nashine
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.
authors.forename: Marilyn
authors.initials: M
authors.lastname: Chwa
authors.affiliation: College of Osteopathic Medicine, Touro University Nevada, Nevada, United States of America.
authors.forename: Mina
authors.initials: M
authors.lastname: Kazemian
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.
authors.forename: Kunal
authors.initials: K
authors.lastname: Thaker
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.|VA Medical Center Long Beach Hospital, Long Beach, California, United States of America.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Lu
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.|Cedars-Sinai Medical Center, Los Angeles, California, United States of America.
authors.forename: Anthony
authors.initials: A
authors.lastname: Nesburn
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.
authors.forename: Baruch D
authors.initials: BD
authors.lastname: Kuppermann
authors.affiliation: Gavin Herbert Eye Institute, University of California Irvine, Irvine, California, United States of America.|Department of Pathology and Laboratory Medicine, University of California Irvine, Irvine, California, United States of America.
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Kenney
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D016022:Case-Control Studies; D002478:Cells, Cultured; D003171:Complement Pathway, Classical / Q000235:genetics*; D003165:Complement System Proteins / Q000235:genetics* / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000235:genetics; D004734:Energy Metabolism; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D054629:Genome, Mitochondrial; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism*; D008268:Macular Degeneration / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics
chemical_list: D015415:Biomarkers; D004272:DNA, Mitochondrial; D003165:Complement System Proteins