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PMID:23152892

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Citation

Xu, XL, Ma, W, Zhu, YB, Wang, C, Wang, BY, An, N, An, L, Liu, Y, Wu, ZH and Tian, JH (2012) The Microtubule-Associated Protein ASPM Regulates Spindle Assembly and Meiotic Progression in Mouse Oocytes. PLoS ONE 7:e49303

Abstract

The microtubule-associated protein ASPM (abnormal spindle-like microcephaly-associated) plays an important role in spindle organization and cell division in mitosis and meiosis in lower animals, but its function in mouse oocyte meiosis has not been investigated. In this study, we characterized the localization and expression dynamics of ASPM during mouse oocyte meiotic maturation and analyzed the effects of the downregulation of ASPM expression on meiotic spindle assembly and meiotic progression. Immunofluorescence analysis showed that ASPM localized to the entire spindle at metaphase I (MI) and metaphase II (MII), colocalizing with the spindle microtubule protein acetylated tubulin (Ac-tubulin). In taxol-treated oocytes, ASPM colocalized with Ac-tubulin on the excessively polymerized microtubule fibers of enlarged spindles and the numerous asters in the cytoplasm. Nocodazole treatment induced the gradual disassembly of microtubule fibers, during which ASPM remained colocalized with the dynamic Ac-tubulin. The downregulation of ASPM expression by a gene-specific morpholino resulted in an abnormal meiotic spindle and inhibited meiotic progression; most of the treated oocytes were blocked in the MI stage with elongated meiotic spindles. Furthermore, coimmunoprecipitation combined with mass spectrometry and western blot analysis revealed that ASPM interacted with calmodulin in MI oocytes and that these proteins colocalized at the spindle. Our results provide strong evidence that ASPM plays a critical role in meiotic spindle assembly and meiotic progression in mouse oocytes.

Links

PubMed Online version:10.1371/journal.pone.0049303

Keywords


Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:ASPM

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ASPM

located_in

GO:0097431: mitotic spindle pole

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ASPM

located_in

GO:0072687: meiotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ASPM

located_in

GO:0005874: microtubule

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(CL:0000023)

Seeded From UniProt

complete

MOUSE:ASPM

acts_upstream_of_or_within

GO:0090306: meiotic spindle assembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

  • occurs_in:(CL:0000023)

Seeded From UniProt

complete

MOUSE:ASPM

GO:0060236: regulation of mitotic spindle organization

ECO:0000314:

P

Figure 4.shows "Downregulation of ASPM expression by morpholinos disrupts meiotic spindle organization."

complete
CACAO 6033

MOUSE:ASPM

GO:0051445: regulation of meiotic cell cycle

ECO:0000315:

P

Table 1 shows meiotic progression.

complete
CACAO 6050

MOUSE:ASPM

enables

GO:0005516: calmodulin binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

MOUSE:ASPM

part_of

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ASPM

part_of

GO:0005874: microtubule

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(CL:0000023)

Seeded From UniProt

complete

MOUSE:ASPM

part_of

GO:0072687: meiotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ASPM

acts_upstream_of_or_within

GO:0090306: spindle assembly involved in meiosis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

occurs_in:(CL:0000023)

Seeded From UniProt

complete

MOUSE:ASPM

part_of

GO:0097431: mitotic spindle pole

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:ASPM

involved_in

GO:0051445: regulation of meiotic cell cycle

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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