Real-time imaging of actin filaments in the zebrafish oocyte and embryo.
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 the analysis of gamete formation and early embryonic development. Here, we report the 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 the 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 the dynamics of actin filaments in oogenesis and early embryogenesis.
Biosystems Science Course, Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
Yumiko
Y
Nukada
Biosystems Science Course, Graduate School of Life Science, Hokkaido University, Sapporo, Japan.
Mayu
M
Horie
Laboratory of Tissue and Polymer Sciences, Faculty of Advanced Life Science, Hokkaido University, Sapporo, Japan.
Akimasa
A
Fukui
Department of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, Japan.
Tomoya
T
Kotani
Department of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, Japan.
Masakane
M
Yamashita
D008841:Actin Cytoskeleton / Q000737:chemistry*; D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D056744:Cyclin B1 / Q000235:genetics; D003571:Cytochalasin B / Q000737:chemistry; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D005260:Female; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008164:Luminescent Proteins; D008297:Male; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D009866:Oogenesis; D011401:Promoter Regions, Genetic; D013737:Testis / Q000378:metabolism; D015027:Zebrafish; D000097573:Red Fluorescent Protein
D000199:Actins; D056744:Cyclin B1; D003571:Cytochalasin B; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008840:Microfilament Proteins; D000097573:Red Fluorescent Protein; C071058:moesin
live imaging; oocyte maturation; oogenesis; transgenic fish; vertebrate