Localization of Mitochondrial Nucleoids by Transmission Electron Microscopy Using the Transgenic Expression of the Mitochondrial Helicase Twinkle and APEX2.
Animals
Mice
Humans
Hydrogen Peroxide
/ metabolism
Mitochondria
/ metabolism
DNA, Mitochondrial
/ genetics
Animals, Genetically Modified
DNA Helicases
/ metabolism
Microscopy, Electron, Transmission
Mitochondrial Proteins
/ metabolism
Endonucleases
/ metabolism
DNA-(Apurinic or Apyrimidinic Site) Lyase
/ metabolism
Multifunctional Enzymes
APEX2
Mitochondria
Nucleoid
TEM
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2023
2023
Historique:
entrez:
22
2
2023
pubmed:
23
2
2023
medline:
25
2
2023
Statut:
ppublish
Résumé
Reminiscent of their evolutionary origin, mitochondria contain their own genome (mtDNA) compacted into the mitochondrial chromosome or nucleoid (mt-nucleoid). Many mitochondrial disorders are characterized by disruption of mt-nucleoids, either by direct mutation of genes involved in mtDNA organization or by interfering with other vital proteins for mitochondrial function. Thus, changes in mt-nucleoid morphology, distribution, and structure are a common feature in many human diseases and can be exploited as an indicator of cellular fitness. Electron microscopy provides the highest possible resolution that can be achieved, delivering spatial and structural information about all cellular structures. Recently, the ascorbate peroxidase APEX2 has been used to increase transmission electron microscopy (TEM) contrast by inducing diaminobenzidine (DAB) precipitation. DAB has the ability to accumulate osmium during classical EM sample preparation and, due to its high electron density, provides strong contrast for TEM. Among the nucleoid proteins, the mitochondrial helicase Twinkle fused with APEX2 has been successfully used to target mt-nucleoids, providing a tool to visualize these subcellular structures with high contrast and with the resolution of an electron microscope. In the presence of H
Identifiants
pubmed: 36807792
doi: 10.1007/978-1-0716-2922-2_13
doi:
Substances chimiques
Hydrogen Peroxide
BBX060AN9V
DNA, Mitochondrial
0
DNA Helicases
EC 3.6.4.-
Mitochondrial Proteins
0
APEX2 protein, human
EC 4.2.99.18
Endonucleases
EC 3.1.-
DNA-(Apurinic or Apyrimidinic Site) Lyase
EC 4.2.99.18
Multifunctional Enzymes
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
173-188Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
Références
Stewart JB, Chinnery PF (2020) Extreme heterogeneity of human mitochondrial DNA from organelles to populations. Nat Rev Genet 22:106. https://doi.org/10.1038/s41576-020-00284-x
doi: 10.1038/s41576-020-00284-x
pubmed: 32989265
Kukat C, Wurm CA, Spahr H, Falkenberg M, Larsson NG, Jakobs S (2011) Super-resolution microscopy reveals that mammalian mitochondrial nucleoids have a uniform size and frequently contain a single copy of mtDNA. Proc Natl Acad Sci U S A 108(33):13534–13539. https://doi.org/10.1073/pnas.1109263108
doi: 10.1073/pnas.1109263108
pubmed: 21808029
pmcid: 3158146
Peter B, Falkenberg M (2020) TWINKLE and other human mitochondrial DNA helicases: structure, function and disease. Genes (Basel) 11(4):doi:10.3390/genes11040408
doi: 10.3390/genes11040408
Tyynismaa H, Sembongi H, Bokori-Brown M, Granycome C, Ashley N, Poulton J, Jalanko A, Spelbrink JN, Holt IJ, Suomalainen A (2004) Twinkle helicase is essential for mtDNA maintenance and regulates mtDNA copy number. Hum Mol Genet 13(24):3219–3227. https://doi.org/10.1093/hmg/ddh342
doi: 10.1093/hmg/ddh342
pubmed: 15509589
Nass MMK, Nass S (1963) Intramitochondrial fibers with DNA characteristics. 1. Fixation and electron staining reactions. J Cell Biol 19(3):593-&. https://doi.org/10.1083/jcb.19.3.593
doi: 10.1083/jcb.19.3.593
pubmed: 14086138
pmcid: 2106331
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313(5793):1642–1645. https://doi.org/10.1126/science.1127344
doi: 10.1126/science.1127344
pubmed: 16902090
Yip KM, Fischer N, Paknia E, Chari A, Stark H (2020) Atomic-resolution protein structure determination by cryo-EM. Nature 587(7832):157–161. https://doi.org/10.1038/s41586-020-2833-4
doi: 10.1038/s41586-020-2833-4
pubmed: 33087927
Faulk WP, Taylor GM (1971) An immunocolloid method for the electron microscope. Immunochemistry 8(11):1081–1083. https://doi.org/10.1016/0019-2791(71)90496-4
doi: 10.1016/0019-2791(71)90496-4
pubmed: 4110101
Iborra FJ, Kimura H, Cook PR (2004) The functional organization of mitochondrial genomes in human cells. BMC Biol 2:9. https://doi.org/10.1186/1741-7007-2-9
doi: 10.1186/1741-7007-2-9
pubmed: 15157274
pmcid: 425603
Schwarz H, Humbel BM (1989) Influence of fixatives and embedding media on Immunolabelling of freeze-substituted cells. Scanning Microsc 3:57–64
Tokuyasu KT (1973) A technique for ultracryotomy of cell suspensions and tissues. J Cell Biol 57(2):551–565. https://doi.org/10.1083/jcb.57.2.551
doi: 10.1083/jcb.57.2.551
pubmed: 4121290
pmcid: 2108989
Allen DE, Perrin DD (1979) Electron cytochemical stains based on metal chelation. Int Rev Cytol 61:63–84. https://doi.org/10.1016/s0074-7696(08)61995-6
doi: 10.1016/s0074-7696(08)61995-6
pubmed: 92462
Graham RC Jr, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14(4):291–302. https://doi.org/10.1177/14.4.291
doi: 10.1177/14.4.291
pubmed: 5962951
Li JL, Wang Y, Chiu SL, Cline HT (2010) Membrane targeted horseradish peroxidase as a marker for correlative fluorescence and electron microscopy studies. Front Neural Circuit 4:6. https://doi.org/10.3389/neuro.04.006.2010
doi: 10.3389/neuro.04.006.2010
Shu X, Lev-Ram V, Deerinck TJ, Qi Y, Ramko EB, Davidson MW, Jin Y, Ellisman MH, Tsien RY (2011) A genetically encoded tag for correlated light and electron microscopy of intact cells, tissues, and organisms. PLoS Biol 9(4):e1001041. https://doi.org/10.1371/journal.pbio.1001041
doi: 10.1371/journal.pbio.1001041
pubmed: 21483721
pmcid: 3071375
Martell JD, Deerinck TJ, Sancak Y, Poulos TL, Mootha VK, Sosinsky GE, Ellisman MH, Ting AY (2012) Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy. Nat Biotechnol 30(11):1143. https://doi.org/10.1038/nbt.2375
doi: 10.1038/nbt.2375
pubmed: 23086203
pmcid: 3699407
Lam SS, Martell JD, Kamer KJ, Deerinck TJ, Ellisman MH, Mootha VK, Ting AY (2015) Directed evolution of APEX2 for electron microscopy and proximity labeling. Nat Methods 12(1):51–54. https://doi.org/10.1038/nmeth.3179
doi: 10.1038/nmeth.3179
pubmed: 25419960
Han S, Udeshi ND, Deerinck TJ, Svinkina T, Ellisman MH, Carr SA, Ting AY (2017) Proximity biotinylation as a method for mapping proteins associated with mtDNA in living cells. Cell Chem Biol 24(3):404–414. https://doi.org/10.1016/j.chembiol.2017.02.002
doi: 10.1016/j.chembiol.2017.02.002
pubmed: 28238724
pmcid: 5886301
Wang LJ, Hsu T, Lin HL, Fu CY (2020) Drosophila MICOS knockdown impairs mitochondrial structure and function and promotes mitophagy in muscle tissue. Biol Open 9(12). https://doi.org/10.1242/bio.054262
Morgenstern JP, Land H (1990) Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Nucleic Acids Res 18(12):3587–3596. https://doi.org/10.1093/nar/18.12.3587
doi: 10.1093/nar/18.12.3587
pubmed: 2194165
pmcid: 331014
Naviaux RK, Costanzi E, Haas M, Verma IM (1996) The pCL vector system: rapid production of helper-free, high-titer, recombinant retroviruses. J Virol 70(8):5701–5705. https://doi.org/10.1128/JVI.70.8.5701-5705.1996
doi: 10.1128/JVI.70.8.5701-5705.1996
pubmed: 8764092
pmcid: 190538
Prieto J, Leon M, Ponsoda X, Sendra R, Bort R, Ferrer-Lorente R, Raya A, Lopez-Garcia C, Torres J (2016) Early ERK1/2 activation promotes DRP1-dependent mitochondrial fission necessary for cell reprogramming. Nat Commun 7:11124. https://doi.org/10.1038/ncomms11124
doi: 10.1038/ncomms11124
pubmed: 27030341
pmcid: 4821885