Analysis of ER-mitochondria contacts using correlative fluorescence microscopy and soft X-ray tomography of mammalian cells

作者：Elgass Kirstin D; Smith Elizabeth A; LeGros Mark A; Larabell Carolyn A^*; Ryan Michael T

来源：Journal of Cell Science, 2015, 128(15): 2795-2804.

DOI：10.1242/jcs.169136

• Confocal microscopy
• Endoplasmic reticulum
• Microscopy imaging
• Mitochondria
• Mitochondrial fission
• Soft X-ray tomography
• Correlative imaging

Mitochondrial fission is important for organelle transport, quality control and apoptosis. Changes to the fission process can result in a wide variety of neurological diseases. In mammals, mitochondrial fission is executed by the GTPase dynamin-related protein 1 (Drp1; encoded by DNM1L), which oligomerizes around mitochondria and constricts the organelle. The mitochondrial outer membrane proteins Mff, MiD49 (encoded by MIEF2) and MiD51 (encoded by MIEF1) are involved in mitochondrial fission by recruiting Drp1 from the cytosol to the organelle surface. In addition, endoplasmic reticulum (ER) tubules have been shown to wrap around and constrict mitochondria before a fission event. Up to now, the presence of MiD49 and MiD51 at ER-mitochondrial division foci has not been established. Here, we combine confocal live-cell imaging with correlative cryogenic fluorescence microscopy and soft x-ray tomography to link MiD49 and MiD51 to the involvement of the ER in mitochondrial fission. We gain further insight into this complex process and characterize the 3D structure of ER-mitochondria contact sites.

• 出版日期2015-8-1
• 单位河北医科大学