Freeze-fracture analysis of isolated liver mitochondria in different metabolic states.
Isolated liver mitochondria were crosslinked with glutaraldehyde during different respiratory states and then analyzed either by freeze-fracturing, or in thin sections prepared according to a low denaturation embedding technique. In freeze-fractured material, large intracristal spaces were found in 86% of the mitochondria freshly isolated in a sucrose medium. These results were in good agreement with mitochondria analyzed in thin sections, in which case 88% of the mitochondria contained intracristal spaces. In freeze-fractured mitochondria, only 24% of the mitochondria crosslinked after 3 min in respiratory state 4 contained intracristal spaces, while 36% of the mitochondria contained intracristal spaces during state 3. These values compared favorably with those obtained in embedded material in which case 21 and 41% of the mitochondria contained intracristal spaces during states 3 and 4, respectively. The agreement between the observations made with the two methods demonstrated that the disappearance of intracristal spaces is independent of the embedding technique, and is not a special effect caused by dehydration in ethylene glycol.
D000818:Animals; D003451:Cryoprotective Agents; D019855:Ethylene Glycol; D005026:Ethylene Glycols; D005614:Freeze Fracturing; D008854:Microscopy, Electron; D008930:Mitochondria, Liver / Q000378:metabolism / Q000648:ultrastructure*; D051381:Rats
D003451:Cryoprotective Agents; D005026:Ethylene Glycols; D019855:Ethylene Glycol