The endoplasmic reticulum (ER) is a large multifunctional organelle of eukaryotic cells. Malfunction of the ER in various disease states, such as atherosclerosis, Type 2 diabetes, cancer, Alzheimer’s and Parkinson’s diseases and amyotrophic lateral sclerosis, often correlates with alterations in its morphology. The ER exhibits regionally variable membrane morphology that includes, at the extremes, large relatively flat surfaces and interconnected tubular structures highly curved in cross-section. Much evidence suggests that ER morphology is controlled by shaping proteins that associate with membrane lipids. To investigate the role of these lipids in ER morphology, we developed a sea urchin oocyte model which is a relatively quiescent cell in which the ER consists mostly of tubules. We altered levels of endogenous diacylglycerol, phosphatidylethanolamine and phosphatidylcholine by microinjection of enzymes or lipid delivery by fusion with liposomes and evaluated shape changes with two- and three-dimensional confocal imaging and three-dimensional electron microscopy techniques. Decreases and increases in the levels of lipids such as diacylglycerol or phosphatidylethanolamine characterized by negative spontaneous curvature correlated with conversion to sheet structures or tubules respectively. The effects of endogenous alterations of diacylglycerol were reversible upon exogenous delivery of lipids of negative spontaneous curvature. These data suggest that shaping proteins require threshold amounts of such lipids and that localized deficiencies of the lipids could contribute to alterations of ER morphology. The oocyte modeling system should be beneficial to future studies directed at understanding the precise spatial and compositional requirements of lipid species in interactions leading to alterations of organelle shaping.
- membrane lipids
- negative spontaneous curvature
- SBF scanning electron tomography
- confocal microscopy
- Electron microscopy
- Endoplasmic reticulum
- Lipid Kinases