Structural and functional relationships between Ca2+ puffs and mitochondria in Xenopus oocytes.
Ca2+ uptake and release from endoplasmic reticulum (ER) and mitochondrial Ca2+ stores play important physiological and pathological roles, and these processes are shaped by interactions that depend on the structural intimacy between these organelles. Here we investigate the morphological and functional relationships between mitochondria, ER, and the sites of intracellular Ca2+ release in Xenopus laevis oocytes by combining confocal imaging of local Ca2+ release events ("Ca2+ puffs") with mitochondrial localization visualized using vital dyes and subcellularly targeted fluorescent proteins. Mitochondria and ER are localized in cortical bands approximately 6-8 microm wide, with the mitochondria arranged as densely packed "islands" interconnected by discrete strands. The ER is concentrated more superficially than mitochondria, and the mean separation between Ca2+ puff sites and mitochondria is approximately 2.3 microm. However, a subpopulation of Ca2+ puff sites is intimately associated with mitochondria (approximately 28% within <600 nm), a greater number than expected if Ca2+ puff sites were randomly distributed. Ca2+ release sites close to mitochondria exhibit lower Ca2+ puff activity than Ca2+ puff sites in regions with lower mitochondrial density. Furthermore, Ca2+ puff sites in close association with mitochondria rarely serve as the sites for Ca2+ wave initiation. We conclude that mitochondria play important roles in regulating local ER excitability, Ca2+ wave initiation, and, thereby, spatial patterning of global Ca2+ signals.
Marchant
Jonathan S
JS
Laboratory of Cellular and Molecular Neurobiology, Department of Neurobiology and Behavior, University of California, Irvine, California 92697-4550, USA.
Ramos
Viviana
V
Parker
Ian
I
D016475:3T3 Cells; D000818:Animals; D020013:Calcium Signaling / Q000502:physiology*; D004721:Endoplasmic Reticulum / Q000378:metabolism / Q000648:ultrastructure; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D018613:Microscopy, Confocal; D018715:Microscopy, Video; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D014982:Xenopus laevis