Real‐time bioluminescence imaging of a protein secretory pathway in living mammalian cells using Gaussia luciferase

Abstract

Using photon counting and charge-coupled device (CCD) cameras, we have applied the method of real-time bioluminescence imaging to investigate protein trafficking in mammalian cells. In the living cells of Chinese hamster ovary and PC12D cells, exocytotic secretion of protein and protein targeting on the cell surface were visualized using the secreted Gaussia luciferase (GLase) as a reporter protein in a minute. After incubation of the cells with luciferin (coelenterazine) for 10 min, luciferin was imported into the cells and the vesicle transport network in the cells could be shown by luminescence images of GLase activity. Further, we demonstrate that GLase with a heterologous signal peptide sequence is targeted to the cell surface in neuronally differentiated PC12D cells and luminescence signals could be detected in a few seconds.

Bioluminescence imaging: progress and applications

“…Upon interfering with the secretory pathway or induction of ER stress, the level of Gluc secreted in the conditioned medium is reduced significantly. The same approach was used by Suzuki et al ., to monitor protein trafficking in mammalian cells [45]. …”

Development of a bifunctional immunoliposome system for combined drug delivery and imaging in vivo

“…As a coelenterazine-dependent luciferase, Gaussia luciferase (GLase) was cloned from a marine copepod Gaussia princes and has recently been validated as a reporter gene for in vivo imaging applications [18,19]. Its small size (19.9 kDa) and independence of ATP make it suitable for detecting bioluminescence when fused with another protein [20][21][22].…”

Bioluminescence Assays for Monitoring Chondrogenic Differentiation and Cartilage Regeneration

“…The visualization of secretion of Cypridina luciferase (Cluc) and Gaussia luciferase (GLuc) was realized in mammalian cells, showing that the secreted luciferases are secreted via the constitutive exocytotic pathways [73,74]. Cluc was thus utilized for imaging neurotransmitter release [75]. …”