PUBLICATION
Examining rhodopsin retention in endoplasmic reticulum and intracellular localization in vitro and in vivo by using truncated rhodopsin fragments
- Authors
- Chen, Y.F., Wang, I.J., Lin, L.L., and Chen, M.S.
- ID
- ZDB-PUB-101201-15
- Date
- 2011
- Source
- Journal of cellular biochemistry 112(2): 520-530 (Journal)
- Registered Authors
- Wang, I-Jong
- Keywords
- rhodopsin, zebrafish, retina
- MeSH Terms
-
- Animals
- COS Cells
- Chlorocebus aethiops
- Endoplasmic Reticulum/metabolism*
- Humans
- Immunohistochemistry
- Microscopy, Confocal
- Point Mutation
- Retina/metabolism*
- Rhodopsin/genetics
- Rhodopsin/metabolism*
- Zebrafish
- PubMed
- 21268073 Full text @ J. Cell. Biochem.
Citation
Chen, Y.F., Wang, I.J., Lin, L.L., and Chen, M.S. (2011) Examining rhodopsin retention in endoplasmic reticulum and intracellular localization in vitro and in vivo by using truncated rhodopsin fragments. Journal of cellular biochemistry. 112(2):520-530.
Abstract
More than 100 mutations of rhodopsin have been identified to be associated with retinitis pigmentosa (RP), and mostly autosomal dominant RP (ADRP). The majority of rhodopsin-associated ADRP is caused by protein misfolding and ER retention. In this study, we aimed to evaluate rhodopsin folding, exiting the ER and intracellular localization through expression of the rhodopsin fragments in COS-1 cells as well as in the transgenic zebrafish. We cloned human rhodopsin cDNA, which was then divided into the N-terminal domain, the C-terminal domain, and the fragment between the N- and C-terminal domains, and examine their intracellular expression in vitro and in vivo. We introduced a point mutation, either F45L or G51V, into this fragment and observed the intracellular localization of these mutants in COS-1 cells and in the zebrafish. The results revealed all of the truncated rhodopsin fragments except for the C-terminal domain and the full-length rhodopsin which had some plasma membrane localization, formed aggregates nearby or within the ER in COS-1 cells; however, the N-terminally truncated rhodopsin fragment, the C-terminal domain, and the full-length rhodopsin could traffic to the ROS in the zebrafish. Besides, the F45L mutation and the G51Vmutation in the rhodopsin fragment between the N- and C-terminal domains produced different effects on the aggresome formation and the intracellular distribution of the mutants both in vivo and in vitro. This current study provides new information about the mutant rhodopsin as well as in treatment of the RP in humans in the future.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping