Golgi Apparatus Structure

The Golgi apparatus, also known as the Golgi complex or Golgi, is found in most eukaryotic cells. The Golgi is a flattened membrane-enclosed organelle that consists of multiple compartments and whose function is a continuation from the ER. Macromolecules such as proteins, lipids, and carbohydrates released from the ER are transported to the cis-Golgi compartment of the Golgi complex [1,2]. These macromolecules continue to be transported and processing through the Golgi stack (which contains medial and trans sub-compartments), then travel to the trans-Golgi compartment where they are packaged into vesicles for either secretion from the cell or for use by other organelles within the cell [1,2].

CellLight fluorescent fusion proteins are useful for the identification and demarcation of the Golgi complex in live cells and to follow the dynamics of intracellular behavior, such as protein trafficking. CellLight Golgi-GFP (Figure 1) and CellLight Golgi-RFP (Figure 2) are ready-to-use constructs that use a human Golgi-resident enzyme, N-acetylgalactosaminyltransferase-2 (12.6 kDa), to target the Golgi complex.

Introducing CellLight fluorescent fusion proteins involves a simple transfection step, using the BacMam technology, and they work like cell stains with minimal toxicity or chemical disruption. These Golgi fusion proteins are compatible with other fluorescent probes for multiplex analysis in live cells, or after formaldehyde fixation for colocalization studies.

Learn more about these and other CellLight fluorescent proteins

Fluorescent ceramides are used extensively to study lipid metabolism and trafficking. They provide selective staining for the Golgi complex in live cells and can also be fixed. NBD dye conjugates are weakly fluorescent in aqueous solvents, and its fluorescence increases in a non-polar environment. The BODIPY dye conjugates (Figures 3 and 4) are brighter and more photostable than the NBD derivative, with less environmental sensitivity. Complexing fluorescent lipids with bovine serum albumin (BSA) eliminates the need for organic solvents to dissolve the lipophilic probe—the BSA-complexed probe can be directly dissolved in water and loaded into live cells.

Fluorescent ceramides can be used in various applications including markers for structural identification of the Golgi complex, outlining boundaries to observe morpho-genetic movements, tracing and tracking of lipid metabolism in cells, and measuring lipid synthesis.

Learn more about Golgi apparatus stains

References

1. Tachikawa M and Mochizuki A (2017). Proc Natl Acad Sci U S A. 114(20):5177-5182. PMID: 28461510.
2. Cooper GM (2000). The Cell: A Molecular Approach. 2nd edition. Sunderland (MA): Sinauer Associates; 2000. The Golgi Apparatus.