PUBLICATION

Real-time imaging of actin filaments in the zebrafish oocyte and embryo

Authors
Nukada, Y., Horie, M., Fukui, A., Kotani, T., Yamashita, M.
ID
ZDB-PUB-150904-1
Date
2015
Source
Cytoskeleton (Hoboken, N.J.)   72(9): 491-501 (Journal)
Registered Authors
Kotani, Tomoya
Keywords
live imaging, oocyte maturation, oogenesis, transgenic fish, vertebrate
MeSH Terms
  • Actin Cytoskeleton/chemistry*
  • Actins/chemistry*
  • Actins/metabolism
  • Animals
  • Animals, Genetically Modified
  • Cyclin B1/genetics
  • Cytochalasin B/chemistry
  • Cytoplasm/metabolism
  • Cytoskeleton/metabolism
  • Embryo, Nonmammalian/metabolism*
  • Female
  • Green Fluorescent Proteins/metabolism
  • Luminescent Proteins
  • Male
  • Microfilament Proteins/genetics
  • Microfilament Proteins/metabolism
  • Microscopy, Fluorescence
  • Oocytes/cytology
  • Oocytes/metabolism*
  • Oogenesis
  • Promoter Regions, Genetic
  • Testis/metabolism
  • Zebrafish
PubMed
26335601 Full text @ Cytoskeleton
Abstract
Dynamic changes of cytoplasmic and cortical actin filaments drive various cellular and developmental processes. Although real-time imaging of actin filaments in living cells has been developed, imaging of actin filaments in specific cells of living organisms remains limited, particularly for analysis of gamete formation and early embryonic development. Here, we report production of transgenic zebrafish expressing the C-terminus of Moesin, an actin filament-binding protein, fused with green fluorescent protein or red fluorescent protein (GFP/RFP-MoeC), under the control of a cyclin B1 promoter. GFP/RFP-MoeC was expressed maternally, which labels the cortical actin cytoskeleton of blastula-stage cells. High levels of GFP/RFP fluorescence were detected in the adult ovary and testis. In the ovaries, GFP/RFP-MoeC was expressed in oocytes but not in follicle cells, which allows us to clearly visualize the organization of actin filaments in different stages of the oocyte. Using full-grown oocytes, we revealed dynamic changes of actin columns assembled in the cortical cytoplasm during oocyte maturation. The number of columns slightly decreased in the early period before germinal vesicle breakdown (GVBD) and then significantly decreased at GVBD, followed by recovery after GVBD. Our transgenic fish are useful for analyzing dynamics of actin filaments in oogenesis and early embryogenesis. This article is protected by copyright. All rights reserved.
Genes / Markers
Figures
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Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes