title: Isolation of mitochondria-associated membranes and mitochondria from animal tissues and cells.
abstract: Many cellular processes require the proper cooperation between mitochondria and the endoplasmic reticulum (ER). Several recent works show that their functional interactions rely on dynamic structural contacts between both organelles. Such contacts, called mitochondria-associated membranes (MAMs), are crucial for the synthesis and intracellular transport of phospholipids, as well as for intracellular Ca(2+) signaling and for the determination of mitochondrial structure. Although several techniques are available to isolate mitochondria, only few are specifically tuned to the isolation of MAM, containing unique regions of ER membranes attached to the outer mitochondrial membrane and mitochondria without contamination from other organelles (i.e., pure mitochondria). Here we provide optimized protocols to isolate these fractions from tissues and cells. These procedures require 4-5 h and can be easily modified and adapted to different tissues and cell types.
authors.affiliation: Department of Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland. m.wieckowski@nencki.gov.pl
authors.forename: Mariusz R
authors.initials: MR
authors.lastname: Wieckowski
authors.forename: Carlotta
authors.initials: C
authors.lastname: Giorgi
authors.forename: Magdalena
authors.initials: M
authors.lastname: Lebiedzinska
authors.forename: Jerzy
authors.initials: J
authors.lastname: Duszynski
authors.forename: Paolo
authors.initials: P
authors.lastname: Pinton
mesh_terms: D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000648:ultrastructure*; D006801:Humans; D007425:Intracellular Membranes; D008928:Mitochondria / Q000648:ultrastructure*; D051336:Mitochondrial Membranes; D013048:Specimen Handling / Q000379:methods