Dynamic distribution of spindlin in nucleoli, nucleoplasm and spindle from primary oocytes to mature eggs and its critical function for oocyte-to-embryo transition in gibel carp.
Spindlin (Spin) was thought as a maternal-effect factor associated with meiotic spindle. Its role for the oocyte-to-embryo transition was suggested in mouse, but its direct evidence for the function had been not obtained in other vertebrates. In this study, we used the CagSpin-specific antibody to investigate CagSpin expression pattern and distribution during oogenesis of gibel carp (Carassius auratus gibelio). First, the oocyte-specific expression pattern and dynamic distribution was revealed in nucleoli, nucleoplasm, and spindle from primary oocytes to mature eggs by immunofluorescence localization. In primary oocytes and growth stage oocytes, CagSpin accumulates in nucleoli in increasing numbers along with the oocyte growth, and its disassembly occurs in vitellogenic oocytes, which implicates that CagSpin may be a major component of a large number of nucleoli in fish growth oocytes. Then, co-localization of CagSpin and β-tubulin was revealed in meiotic spindle of mature egg, indicating that CagSpin is one spindle-associated factor. Moreover, microinjection of CagSpin-specific antibody into the fertilized eggs blocked the first cleavage, and found that the CagSpin depletion resulted in spindle assembly disturbance. Thereby, our study provided the first direct evidence for the critical oocyte-to-embryo transition function of Spin in vertebrates, and confirmed that Spin is one important maternal-effect factor that participates in oocyte growth, oocyte maturation, and oocyte-to-embryo transition.
Sun
Min
M
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate School of Chinese Academy of Sciences, Wuhan, China.
Li
Zhi
Z
Gui
Jian-Fang
JF
D000818:Animals; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000502:physiology*; D002466:Cell Nucleolus / Q000502:physiology*; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D006054:Goldfish / Q000196:embryology / Q000502:physiology*; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D009865:Oocytes / Q000502:physiology*; D009866:Oogenesis / Q000502:physiology*; D010750:Phosphoproteins / Q000502:physiology*; D008941:Spindle Apparatus / Q000502:physiology; D014404:Tubulin / Q000502:physiology*
D018797:Cell Cycle Proteins; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D014404:Tubulin; C104434:spindlin