Detail of Fig4C_N2_BVRAKO2



Project
Title
Confocal fluorescence images of Blvra–/– MEF cells expressing NIR-GECO1 before ionomycin addition.
Description
Confocal fluorescence images of Blvra–/– MEF cells expressing NIR-GECO1 before ionomycin addition.
Release, Updated
2022-03-31
License
CC BY
Kind
Image data
File Formats
.oib
Data size
41.6 MB

Organism
Mus musculus ( NCBI:txid10090 )
Strain(s)
MEFs
Cell Line
-
Reporter
NIR-GECO1

Datatype
-
Molecular Function (MF)
biliverdin reductase ( GO:0004074 )
Biological Process (BP)
-
Cellular Component (CC)
-
Biological Imaging Method
confocal microscopy ( Fbbi:00000251 )
X scale
1.242 micrometer/pixel
Y scale
1.242 micrometer/pixel
Z scale
1 micrometer/slice
T scale
23 sec per time interval

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Kobachi K, et. al. (2020) Cell Struct Funct., 45(2):131-141.
Related paper(s)

Kenju Kobachi, Sota Kuno, Shinya Sato, Kenta Sumiyama, Michiyuki Matsuda, Kenta Terai (2020) Biliverdin Reductase-A Deficiency Brighten and Sensitize Biliverdin-binding Chromoproteins., Cell structure and function, Volume 45, Number 2, pp. 131-141

Published in 2020 Aug 21 (Electronic publication in June 25, 2020, midnight )

(Abstract) Tissue absorbance, light scattering, and autofluorescence are significantly lower in the near-infrared (NIR) range than in the visible range. Because of these advantages, NIR fluorescent proteins (FPs) are in high demand for in vivo imaging. Nevertheless, application of NIR FPs such as iRFP is still limited due to their dimness in mammalian cells. In contrast to GFP and its variants, iRFP requires biliverdin (BV) as a chromophore. The dimness of iRFP is at least partly due to rapid reduction of BV by biliverdin reductase-A (BLVRA). Here, we established biliverdin reductase-a knockout (Blvra(-/-)) mice to increase the intracellular BV concentration and, thereby, to enhance iRFP fluorescence intensity. As anticipated, iRFP fluorescence intensity was significantly increased in all examined tissues of Blvra(-/-) mice. Similarly, the genetically encoded calcium indicator NIR-GECO1, which is engineered based on another NIR FP, mIFP, exhibited a marked increase in fluorescence intensity in mouse embryonic fibroblasts derived from Blvra(-/-) mice. We expanded this approach to an NIR light-sensing optogenetic tool, the BphP1-PpsR2 system, which also requires BV as a chromophore. Again, deletion of the Blvra gene markedly enhanced the light response in HeLa cells. These results indicate that the Blvra(-/-) mouse is a versatile tool for the in vivo application of NIR FPs and NIR light-sensing optogenetic tools.Key words: in vivo imaging, near-infrared fluorescent protein, biliverdin, biliverdin reductase, optogenetic tool.
(MeSH Terms)

Contact
Kenta Terai , Graduate School of Medicine, Kyoto University , Department of Pathology and Biology of Diseases
Contributors

OMERO Dataset
OMERO Project
Source