An Enhanced Mutant of Red Fluorescent Protein DsRed for Double Labeling and Developmental Timer of Neural Fiber Bundle Formation

Vladislav Verkhusha, Hideo Otsuna, Takeshi Awasaki, Hiroki Oda, Shoichiro Tsukita, Kei Ito

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

We developed a new variant of coral-derived red fluorescent protein, DsRed S197Y, which is brighter and essentially free from secondary fluorescence peak. This makes it an ideal reporter for double labeling with green fluorescent protein (GFP). Though purified protein shows only 20% stronger fluorescence emission, culture cells that express DsRed S197Y exhibit a 3-3.5 times higher level of fluorescence than the cells that express wild-type DsRed. The much slower fluorescence maturation of DsRed than that of GFP is a beneficial feature for a fluorescent developmental timer application. When GFP and DsRed S197Y are expressed simultaneously, emissions start at different latency. This provides information about the time after the onset of expression. It reflects the order of cell differentiation if the expression is activated upon differentiation of certain types of cells. We applied this system to the developing brain of Drosophila and visualized, for the first time, the formation order of neural fibers within a large bundle. Our results showed that newly extending fibers of the mushroom body neurons mainly run into the core rather than to the periphery of the existing bundle. DsRed-based timer thus presents an indispensable tool for developmental biology and genetics of model organisms.

Original languageEnglish (US)
Pages (from-to)29621-29624
Number of pages4
JournalJournal of Biological Chemistry
Volume276
Issue number32
DOIs
StatePublished - Aug 10 2001
Externally publishedYes

Fingerprint

Labeling
Fibers
Green Fluorescent Proteins
Fluorescence
Cell Differentiation
Mushroom Bodies
Developmental Biology
Anthozoa
Genetic Models
Cell culture
Drosophila
Neurons
red fluorescent protein
fluorescent protein 583
Brain
Cell Culture Techniques
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

An Enhanced Mutant of Red Fluorescent Protein DsRed for Double Labeling and Developmental Timer of Neural Fiber Bundle Formation. / Verkhusha, Vladislav; Otsuna, Hideo; Awasaki, Takeshi; Oda, Hiroki; Tsukita, Shoichiro; Ito, Kei.

In: Journal of Biological Chemistry, Vol. 276, No. 32, 10.08.2001, p. 29621-29624.

Research output: Contribution to journalArticle

Verkhusha, Vladislav ; Otsuna, Hideo ; Awasaki, Takeshi ; Oda, Hiroki ; Tsukita, Shoichiro ; Ito, Kei. / An Enhanced Mutant of Red Fluorescent Protein DsRed for Double Labeling and Developmental Timer of Neural Fiber Bundle Formation. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 32. pp. 29621-29624.
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