author_facet Xie, Di
Zhang, Juan
Ding, JinLi
Yang, Jing
Zhang, Yan
Xie, Di
Zhang, Juan
Ding, JinLi
Yang, Jing
Zhang, Yan
author Xie, Di
Zhang, Juan
Ding, JinLi
Yang, Jing
Zhang, Yan
spellingShingle Xie, Di
Zhang, Juan
Ding, JinLi
Yang, Jing
Zhang, Yan
PeerJ
OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
General Agricultural and Biological Sciences
General Biochemistry, Genetics and Molecular Biology
General Medicine
General Neuroscience
author_sort xie, di
spelling Xie, Di Zhang, Juan Ding, JinLi Yang, Jing Zhang, Yan 2167-8359 PeerJ General Agricultural and Biological Sciences General Biochemistry, Genetics and Molecular Biology General Medicine General Neuroscience http://dx.doi.org/10.7717/peerj.8180 <jats:sec> <jats:title>Background</jats:title> <jats:p>OLA1 is a member of the GTPase protein family; unlike other members, it possess both GTPase and ATPase activities, and can bind and hydrolyze ATP more efficiently than GTP. OLA1 participates in cell proliferation, oxidative response, protein synthesis and tumorigenesis. However, whether OLA1 is also required for oocyte meiosis is still unknown.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, the localization, expression, and functions of OLA1 in the mouse oocyte meiosis were examined. Immunofluorescent and confocal microscopy were used to explore the location pattern of OLA1 in the mouse oocyte. Moreover, nocodazole treatment was used to confirm the spindle-like location of OLA1 during mouse meiosis. Western blot was used to explore the expression pattern of OLA1 in the mouse oocyte. Microinjection of siRNA was used to explore the OLA1 functions in the mouse oocyte meiosis. In addition, chromosome spreading was used to investigate the spindle assembly checkpoint (SAC) activity.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>Immunofluorescent staining showed that OLA1 evenly distributed in the cytoplasm at germinal vesicle (GV) stage. After meiosis resumption (GVBD), OLA1 co-localized with spindles, which was further identified by nocodazole treatment experiments. Knockdown of OLA1 impaired the germinal vesicle breakdown progression and finally resulted in a lower polar body extrusion rate. Immunofluorescence analysis indicated that knockdown of OLA1 led to abnormal spindle assembly, which was evidenced by multipolar spindles in OLA1-RNAi-oocytes. After 6 h post-GVBD in culture, an increased proportion of oocyte which has precociously entered into anaphase/telephase I (A/TI) was observed in OLA1-knockdown oocytes, suggesting that loss of OLA1 resulted in the premature segregation of homologous chromosomes. In addition, the chromosome spread analysis suggested that OLA1 knockdown induced premature anaphase onset was due to the precocious inactivation of SAC. Taken together, we concluded that OLA1 plays important role in GVBD, spindle assembly and SAC activation maintenance in oocyte meiosis.</jats:p> </jats:sec> OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes PeerJ
doi_str_mv 10.7717/peerj.8180
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title OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_unstemmed OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_full OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_fullStr OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_full_unstemmed OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_short OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_sort ola1 is responsible for normal spindle assembly and sac activation in mouse oocytes
topic General Agricultural and Biological Sciences
General Biochemistry, Genetics and Molecular Biology
General Medicine
General Neuroscience
url http://dx.doi.org/10.7717/peerj.8180
publishDate 2020
physical e8180
description <jats:sec> <jats:title>Background</jats:title> <jats:p>OLA1 is a member of the GTPase protein family; unlike other members, it possess both GTPase and ATPase activities, and can bind and hydrolyze ATP more efficiently than GTP. OLA1 participates in cell proliferation, oxidative response, protein synthesis and tumorigenesis. However, whether OLA1 is also required for oocyte meiosis is still unknown.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, the localization, expression, and functions of OLA1 in the mouse oocyte meiosis were examined. Immunofluorescent and confocal microscopy were used to explore the location pattern of OLA1 in the mouse oocyte. Moreover, nocodazole treatment was used to confirm the spindle-like location of OLA1 during mouse meiosis. Western blot was used to explore the expression pattern of OLA1 in the mouse oocyte. Microinjection of siRNA was used to explore the OLA1 functions in the mouse oocyte meiosis. In addition, chromosome spreading was used to investigate the spindle assembly checkpoint (SAC) activity.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>Immunofluorescent staining showed that OLA1 evenly distributed in the cytoplasm at germinal vesicle (GV) stage. After meiosis resumption (GVBD), OLA1 co-localized with spindles, which was further identified by nocodazole treatment experiments. Knockdown of OLA1 impaired the germinal vesicle breakdown progression and finally resulted in a lower polar body extrusion rate. Immunofluorescence analysis indicated that knockdown of OLA1 led to abnormal spindle assembly, which was evidenced by multipolar spindles in OLA1-RNAi-oocytes. After 6 h post-GVBD in culture, an increased proportion of oocyte which has precociously entered into anaphase/telephase I (A/TI) was observed in OLA1-knockdown oocytes, suggesting that loss of OLA1 resulted in the premature segregation of homologous chromosomes. In addition, the chromosome spread analysis suggested that OLA1 knockdown induced premature anaphase onset was due to the precocious inactivation of SAC. Taken together, we concluded that OLA1 plays important role in GVBD, spindle assembly and SAC activation maintenance in oocyte meiosis.</jats:p> </jats:sec>
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author Xie, Di, Zhang, Juan, Ding, JinLi, Yang, Jing, Zhang, Yan
author_facet Xie, Di, Zhang, Juan, Ding, JinLi, Yang, Jing, Zhang, Yan, Xie, Di, Zhang, Juan, Ding, JinLi, Yang, Jing, Zhang, Yan
author_sort xie, di
container_start_page 0
container_title PeerJ
container_volume 8
description <jats:sec> <jats:title>Background</jats:title> <jats:p>OLA1 is a member of the GTPase protein family; unlike other members, it possess both GTPase and ATPase activities, and can bind and hydrolyze ATP more efficiently than GTP. OLA1 participates in cell proliferation, oxidative response, protein synthesis and tumorigenesis. However, whether OLA1 is also required for oocyte meiosis is still unknown.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, the localization, expression, and functions of OLA1 in the mouse oocyte meiosis were examined. Immunofluorescent and confocal microscopy were used to explore the location pattern of OLA1 in the mouse oocyte. Moreover, nocodazole treatment was used to confirm the spindle-like location of OLA1 during mouse meiosis. Western blot was used to explore the expression pattern of OLA1 in the mouse oocyte. Microinjection of siRNA was used to explore the OLA1 functions in the mouse oocyte meiosis. In addition, chromosome spreading was used to investigate the spindle assembly checkpoint (SAC) activity.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>Immunofluorescent staining showed that OLA1 evenly distributed in the cytoplasm at germinal vesicle (GV) stage. After meiosis resumption (GVBD), OLA1 co-localized with spindles, which was further identified by nocodazole treatment experiments. Knockdown of OLA1 impaired the germinal vesicle breakdown progression and finally resulted in a lower polar body extrusion rate. Immunofluorescence analysis indicated that knockdown of OLA1 led to abnormal spindle assembly, which was evidenced by multipolar spindles in OLA1-RNAi-oocytes. After 6 h post-GVBD in culture, an increased proportion of oocyte which has precociously entered into anaphase/telephase I (A/TI) was observed in OLA1-knockdown oocytes, suggesting that loss of OLA1 resulted in the premature segregation of homologous chromosomes. In addition, the chromosome spread analysis suggested that OLA1 knockdown induced premature anaphase onset was due to the precocious inactivation of SAC. Taken together, we concluded that OLA1 plays important role in GVBD, spindle assembly and SAC activation maintenance in oocyte meiosis.</jats:p> </jats:sec>
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spelling Xie, Di Zhang, Juan Ding, JinLi Yang, Jing Zhang, Yan 2167-8359 PeerJ General Agricultural and Biological Sciences General Biochemistry, Genetics and Molecular Biology General Medicine General Neuroscience http://dx.doi.org/10.7717/peerj.8180 <jats:sec> <jats:title>Background</jats:title> <jats:p>OLA1 is a member of the GTPase protein family; unlike other members, it possess both GTPase and ATPase activities, and can bind and hydrolyze ATP more efficiently than GTP. OLA1 participates in cell proliferation, oxidative response, protein synthesis and tumorigenesis. However, whether OLA1 is also required for oocyte meiosis is still unknown.</jats:p> </jats:sec> <jats:sec> <jats:title>Methods</jats:title> <jats:p>In this study, the localization, expression, and functions of OLA1 in the mouse oocyte meiosis were examined. Immunofluorescent and confocal microscopy were used to explore the location pattern of OLA1 in the mouse oocyte. Moreover, nocodazole treatment was used to confirm the spindle-like location of OLA1 during mouse meiosis. Western blot was used to explore the expression pattern of OLA1 in the mouse oocyte. Microinjection of siRNA was used to explore the OLA1 functions in the mouse oocyte meiosis. In addition, chromosome spreading was used to investigate the spindle assembly checkpoint (SAC) activity.</jats:p> </jats:sec> <jats:sec> <jats:title>Results</jats:title> <jats:p>Immunofluorescent staining showed that OLA1 evenly distributed in the cytoplasm at germinal vesicle (GV) stage. After meiosis resumption (GVBD), OLA1 co-localized with spindles, which was further identified by nocodazole treatment experiments. Knockdown of OLA1 impaired the germinal vesicle breakdown progression and finally resulted in a lower polar body extrusion rate. Immunofluorescence analysis indicated that knockdown of OLA1 led to abnormal spindle assembly, which was evidenced by multipolar spindles in OLA1-RNAi-oocytes. After 6 h post-GVBD in culture, an increased proportion of oocyte which has precociously entered into anaphase/telephase I (A/TI) was observed in OLA1-knockdown oocytes, suggesting that loss of OLA1 resulted in the premature segregation of homologous chromosomes. In addition, the chromosome spread analysis suggested that OLA1 knockdown induced premature anaphase onset was due to the precocious inactivation of SAC. Taken together, we concluded that OLA1 plays important role in GVBD, spindle assembly and SAC activation maintenance in oocyte meiosis.</jats:p> </jats:sec> OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes PeerJ
spellingShingle Xie, Di, Zhang, Juan, Ding, JinLi, Yang, Jing, Zhang, Yan, PeerJ, OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes, General Agricultural and Biological Sciences, General Biochemistry, Genetics and Molecular Biology, General Medicine, General Neuroscience
title OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_full OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_fullStr OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_full_unstemmed OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_short OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
title_sort ola1 is responsible for normal spindle assembly and sac activation in mouse oocytes
title_unstemmed OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes
topic General Agricultural and Biological Sciences, General Biochemistry, Genetics and Molecular Biology, General Medicine, General Neuroscience
url http://dx.doi.org/10.7717/peerj.8180