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Abstract
Adenosine diphosphate-ribosylation factor 1 (ARF1) is a member of guanosine triphosphate (GTP)-binding proteins family associated with Golgi complexes. ARF1 regulated asymmetric cell division in female meiosis in mouse. However, little isknown about its function in bovine oocyte meiosis. In the present study, we examined the localization, expression and functions of ARF1 during bovine oocyte meiotic maturation. Active form ARF1Q71L-Venus showed that ARF1 co-localized with α-tubulin on the spindle from the M I to the M II stage. Inhibition of ARF1 activity by microinjecting mRNA of a dominant negative mutant form of ARF1 (ARF1T31N ) or ARF1 morpholino (ARF1 MO) into the germinal vesicle (GV) oocytes, and treating GV oocytes with brefeldin A (BFA), an inhibitor of Golgi-based membrane fusion led to abnormal spindle assembly and cytokinesis failure. On the contrary, microinjection of mRNA of a positive mutant form of ARF1 (ARF1Q71L) into GV oocytes had no effect on spindle assembly and the oocytes could undergo normal cytokinesis to generate one large egg with one small polar body. From the above, our results suggest that ARF1 plays an essential role in spindle assembly in bovine oocytes. Key words: Adenosine diphosphate-ribosylation factor 1 (ARF1), spindle assembly, oocytes meiosis cytokinesis, bovine.
Highlights
Female meiotic divisions are asymmetrical and generate a large oocyte and two small polar bodies in mammalians, which is essential to preserve the maternal complement of resources necessary to support subsequent early development and maintain the chromosome number constant (Zheng and Dean, 2009)
By expressing the dominant negative mutant form ARF1T31N or microinjecting of Adenosine diphosphate-ribosylation factor 1 (ARF1) morpholino (ARF1 MO) in germinal vesicle (GV) and treating GV oocytes with brefeldin A (BFA), we found that a large proportion of oocytes could not undergo cytokinesis and the spindles of oocytes were abnormal
To investigate the role of ARF1 in bovine oocyte maturation, reverse transcriptase (RT)-PCR was performed to verify wether ARF1 is expressed in oocyte maturation
Summary
Female meiotic divisions are asymmetrical and generate a large oocyte and two small polar bodies in mammalians, which is essential to preserve the maternal complement of resources necessary to support subsequent early development and maintain the chromosome number constant (Zheng and Dean, 2009). This asymmetry results from the anchoring of the meiotic spindle to the oocyte cortex and subsequent cortical reorganization (Leader et al, 2002; Maro and Verlhac, 2002; Verlhac et al, 2000). ARF1T31N can trigger a brefeldin Alike phenotype resulting in the redistribution of beta coatomer protein (β-COP) from Golgi membranes to the cytosol and the collapse of the Golgi into the ER
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