This article may be too technical for most readers to understand. Please help improve it to make it understandable to non-experts, without removing the technical details. (March 2015) (Learn how and when to remove this message)

Lipid droplets, also referred to as lipid bodies, oil bodies or adiposomes,^[1] are lipid-rich cellular organelles that regulate the storage and hydrolysis of neutral lipids and are found largely in the adipose tissue.^[2] They also serve as a reservoir for cholesterol and acyl-glycerols for membrane formation and maintenance. Lipid droplets are found in all eukaryotic organisms and store a large portion of lipids in mammalian adipocytes. Initially, these lipid droplets were considered to merely serve as fat depots, but since the discovery in the 1990s of proteins in the lipid droplet coat that regulate lipid droplet dynamics and lipid metabolism, lipid droplets are seen as highly dynamic organelles that play a very important role in the regulation of intracellular lipid storage and lipid metabolism. The role of lipid droplets outside of lipid and cholesterol storage has recently begun to be elucidated and includes a close association to inflammatory responses through the synthesis and metabolism of eicosanoids and to metabolic disorders such as obesity, cancer,^[3][4] and atherosclerosis.^[5] In non-adipocytes, lipid droplets are known to play a role in protection from lipotoxicity by storage of fatty acids in the form of neutral triacylglycerol, which consists of three fatty acids bound to glycerol. Alternatively, fatty acids can be converted to lipid intermediates like diacylglycerol (DAG), ceramides and fatty acyl-CoAs. These lipid intermediates can impair insulin signaling, which is referred to as lipid-induced insulin resistance and lipotoxicity.^[6] Lipid droplets also serve as platforms for protein binding and degradation. Finally, lipid droplets are known to be exploited by pathogens such as the hepatitis C virus, the dengue virus and Chlamydia trachomatis among others.^[7][8]

Cells need to adjust the size and structure of their organelles to keep up with growth and changing environmental conditions. To do this, they either make new phospholipids—the main components of organelle membranes—or modify their fatty acid (FA) content. Fatty acids are also used to produce triacylglycerols (TGs), which store energy in structures called lipid droplets. ^[9]The synthesis of triacylglycerols (TG) can be occurred through two different enzyme pathways. Diacylglycerol Acyltransferases (DGATs) like Dga1 are enzymes found in most eukaryotes. They add an acyl group from a fatty acid that has been activated with coenzyme A (FA-CoA) to diacylglycerol (DG), forming TG. Phospholipid-Diacylglycerol Acyltransferases (PDATs) are enzymes primarily found in fungi, microalgae, and plants. PDATs like Lro1 in yeast transfer a fatty acid directly from a phospholipid to DG to form TG. Moreover, Lro1 couple TG synthesis with the deacylation of membrane phospholipids (PL), resulting in the formation of TG and lysophospholipids (LPL).^[10]

When nutrients become available, the yeast cells enter the exponential growth phase (EXP) to grow quickly. It has been shown that during the EXP phase, Lro1-GFP is localized in the endoplasmic reticulum (ER) to synthesize triacylglycerols (TG), which are essential for phospholipid synthesis. ^[11]However, when nutrients become scarce, the cells enter the post-diauxic shift (PDS) phase and Lro1-GFP no longer is in the ER. Instead, it moves to a specific area of the nuclear envelope. This relocation suggests a shift in Lro1's role, possibly in response to the stress of nutrient depletion. Furthermore, this movement is influenced by signals from the cell cycle and nutrient availability, and it stops when the nucleus grows larger.^[12]

Two approaches were used to investigate whether Lro1 can access the inner nuclear membrane (INM). ^[13]In yeast, the ability of integral membrane proteins to move from the endoplasmic reticulum (ER) to the inner nuclear membrane (INM) is restricted by the size of their cytosolic domains. Proteins with cytosolic domains larger than 90 kDa cannot pass through the nuclear pore complex into the INM. It has been shown that when the Lro1’s N-terminal domain was enlarged with one, two, or three copies of the maltose-binding protein (MBP), its ability to target the nucleolus was significantly reduced. This suggests that Lro1 normally resides at the INM, but when its N-domain becomes too large, it can no longer pass through the nuclear pore complex to reach the INM and these larger proteins are likely being degraded.^[14]

The anchor-away technique was used as the second approach. The researchers fused the INM protein Heh1 with FK506 binding protein (FKBP12) to serve as an anchor at the INM and interact with other proteins. Lro1 was fused to GFP for visualization and the FKBP12-rapamycin-binding (FRB) domain. This fusion enables Lro1 to interact with Heh1 at the INM in the presence of rapamycin. FRB-GFP construct contains GFP fused to the FRB domain but without Lro1, to show the specific effects of Lro1’s presence and used as a control. ^[15]Upon the addition of rapamycin, FRB-GFP quickly (within 30 minutes) changed from a diffuse distribution to a ring-like pattern, indicating that it had been recruited to the INM by Heh1-FKBP12. This ring-like localization confirmed that the INM anchor (Heh1) is accessible to FRB-GFP. In the strain expressing FRB-Lro1-GFP, rapamycin treatment caused a loss of Lro1’s cortical ER localization and its accumulation at a perinuclear ring, which is characteristic of INM proteins. This suggests that Lro1, via its N-domain, can indeed associate with the INM. In contrast, when Lro1’s N-terminal domain was enlarged by adding 3xMBP, the fusion protein retained its localization at the cortical ER even after rapamycin treatment. ^[16]

The nuclear membrane near the nucleolus tends to expand when there's excess phospholipid synthesis. It has been investigated that whether the protein Lro1 is catalytically active to produce triacylglycerol (TG) in this specific membrane area.^[17] To test Lro1's activity, the researchers expressed it in a yeast strain that couldn't produce any neutral lipids on its own. They did this by deleting four key enzymes involved in lipid production: the DG acyltransferases (LRO1 and DGA1) and the steryl acyltransferases (ARE1 and ARE2). This mutant strain, called "4D," lacks neutral lipids and lipid droplets (LDs), making it ideal for studying Lro1's function.^[18] The mutant "4D" yeast cells cannot survive under nutrient-poor conditions because they cannot make triacylglycerol (TG) or lipid droplets (LDs), which are essential for survival during this phase. When Lro1 is reintroduced as the only enzyme capable of producing TG, it rescues the cells, allowing them to survive better during the stationary phase by forming lipid droplets. ^[19] Moreover, Lro1 with a mutation in the conserved lipase motif cannot perform its catalytic function, meaning it cannot produce TG. As a result, these cells also fail to survive in PDS, similar to the cells without any functional Lro1, demonstrating that the catalytic activity of Lro1 is crucial for survival and LD formation.^[20]

These findings^[21] suggest that Lro1's activity in the nucleus creates a local site for TG synthesis, which helps reshape the nuclear membrane as needed.^[22]

Lipid droplets are composed of a neutral lipid core consisting mainly of triacylglycerols (TAGs) and cholesteryl esters surrounded by a phospholipid monolayer.^[2] The surface of lipid droplets is decorated by a number of proteins which are involved in the regulation of lipid metabolism.^[2] The first and best-characterized family of lipid droplet coat proteins is the perilipin protein family, consisting of five proteins. These include perilipin 1 (PLIN1), perilipin 2 (PLIN2/ ADRP),^[23] perilipin 3 (PLIN3/ TIP47), perilipin 4 (PLIN4/ S3-12) and perilipin 5 (PLIN5/ OXPAT/ LSDP5/ MLDP).^[24][25][26] Proteomics studies have elucidated the association of many other families of proteins to the lipid surface including proteins involved in membrane trafficking, vesicle docking, endocytosis and exocytosis.^[27] Analysis of the lipid composition of lipid droplets has revealed the presence of a diverse set of phospholipid species;^[28] phosphatidylcholine and phosphatidylethanolamine are the most abundant, followed by phosphatidylinositol.

Lipid droplets vary greatly in size, ranging from 20 to 40 nm to 100 um.^[29] In adipocytes, lipid bodies tend to be larger and they may compose the majority of the cell, while in other cells they may only be induced under certain conditions and are considerably smaller in size.

Lro1 is a type II integral membrane protein. The N-terminal domain facing the cytoplasm or nucleoplasm and containing a short basic region (RKRR). The larger luminal domain contains the catalytic PDAT domain, which is located within the lumen of the endoplasmic reticulum (ER). The N-terminal domain of Lro1 along with the transmembrane segment showed intranuclear localization with clear enrichment at the nucleolus.^[30] When the K/R residues in the N-terminal domain are mutated to alanines, the nucleolar enrichment is partially compromised but not completely lost, indicating that other regions of Lro1 are also contributing to its proper localization. Furthermore, the N-terminal domain of Lro1 was replaced with 4 IgG binding domains of Protein A (4xIgGb). This replacement leads to a loss of localization at both the nucleolus and ER during the PDS phase.^[31]

Lipid droplets bud off the membrane of the endoplasmic reticulum. Initially, a lens is formed by accumulation of TAGs between the two layers of its phospholipid membrane. Nascent lipid droplets may grow by diffusion of fatty acids, endocytosis of sterols, or fusion of smaller lipid droplets through the aid of SNARE proteins.^[29] The budding of lipid droplets is promoted by an asymmetric accumulation of phospholipids that decrease surface tension towards the cytosol.^[32] Lipid droplets have also been observed to be created by the fission of existing lipid droplets, though this is thought to be less common than de novo formation.^[33]

The formation of lipid droplets from the endoplasmic reticulum begins with the synthesis of the neutral lipids to be transported. The manufacture of TAGs from diacylglycerol (by the addition of a fatty acyl chain) is catalyzed by the DGAT proteins, though the extent to which these and other proteins are required depends on cell type.^[34] Neither DGAT1 nor DGAT2 is singularly essential for TAG synthesis or droplet formation, though mammalian cells lacking both cannot form lipid droplets and have severely stunted TAG synthesis. DGAT1, which seems to prefer exogenous fatty acid substrates, is not essential for life; DGAT2, which seems to prefer endogenously synthesized fatty acids, is.^[33]

In non-adipocytes, lipid storage, lipid droplet synthesis and lipid droplet growth can be induced by various stimuli including growth factors, long-chain unsaturated fatty acids (including oleic acid and arachidonic acid), oxidative stress and inflammatory stimuli such bacterial lipopolysaccharides, various microbial pathogens, platelet-activating factor, eicosanoids, and cytokines.^[35]

An example is the endocannabinoids that are unsaturated fatty acid derivatives, which mainly are considered to be synthesised “on demand” from phospholipid precursors residing in the cell membrane, but may also be synthesised and stored in intracellular lipid droplets and released from those stores under appropriate conditions.^[36]

It is possible to observe the formation of lipid droplets, live and label-free, using label-free live-cell imaging.

• 
  Lipid droplets in murine microglia
• 
  Oil bodies in Nardia scalaris, a leafy liverwort

• Oil body
• Adipocyte
• Lipolysis
• Seipin

• Media related to Lipid droplets at Wikimedia Commons