scholarly journals CEP215 and AURKA regulate spindle pole focusing and aMTOC organization in mouse oocytes

Reproduction ◽  
2020 ◽  
Vol 159 (3) ◽  
pp. 261-274
Author(s):  
Xiaotian Wang ◽  
Claudia Baumann ◽  
Rabindranath De La Fuente ◽  
Maria M Viveiros
Keyword(s):  
Critical Role ◽  
Super Resolution ◽  
Aurora Kinase ◽  
Spindle Pole ◽  
Meiotic Spindle ◽  
Dependent Manner ◽  
Aurora Kinase A ◽  
Mouse Oocytes ◽  
Kinase A

Acentriolar microtubule-organizing centers (aMTOCs) play a critical role in stable meiotic spindle assembly in oocytes, necessary for accurate chromosome segregation. Yet, there is a limited understanding of the essential regulatory components of these unique MTOCs. In somatic cells, CEP215 (Centrosomal Protein 215) serves as an important regulator of centrosome maturation and spindle organization. Here, we assessed whether it has a similar function in mouse oocytes. CEP215 was detected in oocyte lysates and specifically localized to aMTOCs throughout the progression of meiosis in a pericentrin-dependent manner. Super-resolution microscopy revealed CEP215 co-localization with pericentrin and a unique pore/ring-like structural organization of aMTOCs. Interestingly, inhibition of Aurora Kinase A in either MI or MII-stage oocytes resulted in a striking loss of the ring-like aMTOC organization and pronounced CEP215 clustering at spindle poles, as well as shorter spindles with highly focused poles. In vitro siRNA-mediated transcript knockdown effectively reduced CEP215 in approximately 85% of the oocytes. Maturation rates to MII were similar in the Cep215 siRNA and injected controls; however, a high percentage (~40%) of the Cep215-knockdown oocytes showed notable variations in spindle pole focusing. Surprisingly, pericentrin and γ-tubulin localization and fluorescence intensity at aMTOCs were unaltered in knockdown oocytes, contrasting with mitotic cells where CEP215 depletion reduced γ-tubulin at centrosomes. Our results demonstrate that CEP215 is a functional component of oocyte aMTOCs and participates in the regulation of meiotic spindle pole focusing. Moreover, these studies reveal a vital role for Aurora Kinase A activity in the maintenance of aMTOC organization in oocytes.

scholarly journals Mechanisms underlying disruption of oocyte spindle stability by bisphenol compounds

Reproduction ◽  
2020 ◽  
Vol 159 (4) ◽  
pp. 383-396 ◽  
Author(s):  
Luhan Yang ◽  
Claudia Baumann ◽  
Rabindranth De La Fuente ◽  
Maria M Viveiros
Keyword(s):  
Aurora Kinase ◽  
Rapid Onset ◽  
Spindle Pole ◽  
Potential Interaction ◽  
Meiotic Spindle ◽  
Dependent Manner ◽  
Aurora Kinase A ◽  
Multipolar Spindles ◽  
Spindle Stability ◽  
Kinase A

Accurate chromosome segregation relies on correct chromosome-microtubule interactions within a stable bipolar spindle apparatus. Thus, exposure to spindle disrupting compounds can impair meiotic division and genomic stability in oocytes. The endocrine disrupting activity of bisphenols such as bisphenol A (BPA) is well recognized, yet their damaging effects on spindle microtubules (MTs) is poorly understood. Here, we tested the effect(s) of acute exposure to BPA and bisphenol F (BPF) on assembled spindle stability in ovulated oocytes. Brief (4 h) exposure to increasing concentrations (5, 25, and 50 µg/mL) of BPA or BPF disrupted spindle organization in a dose-dependent manner, resulting in significantly shorter spindles with highly unfocused poles and fragmented pericentrin. The chromosomes remained congressed in an abnormally elongated metaphase-like configuration, yet normal end-on chromosome-MT attachments were reduced in BPF-treated oocytes. Live-cell imaging revealed a rapid onset of bisphenol-mediated spindle MT disruption that was reversed upon compound removal. Moreover, MT stability and regrowth were impaired in BPA-exposed oocytes, with few cold-stable MTs and formation of multipolar spindles upon MT regrowth. MT-associated kinesin-14 motor protein (HSET/KIFC1) labeling along the spindle was also lower in BPA-treated oocytes. Conversely, cold stable MTs and HSET labeling persisted after BPF exposure. Notably, inhibition of Aurora Kinase A limited bisphenol-mediated spindle pole widening, revealing a potential interaction. These results demonstrate rapid MT disrupting activity by bisphenols, which is highly detrimental to meiotic spindle stability and organization. Moreover, we identify an important link between these defects and altered distribution of key spindle associated factors as well as Aurora Kinase A activity.

scholarly journals Aurora kinase A is essential for meiosis in mouse oocytes

2021 ◽  
Author(s):  
Cecilia S. Blengini ◽  
Patricia Ibrahimian ◽  
Michaela Vaskovicova ◽  
David Drutovic ◽  
Petr Solc ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Aurora Kinase ◽  
The Other ◽  
Meiotic Spindle ◽  
Independent Manner ◽  
Aurora Kinase A ◽  
Mouse Oocytes ◽  
Spindle Poles ◽  
Knockout Model ◽  
Kinase A

The Aurora protein kinases are well-established regulators of spindle building and chromosome segregation in mitotic and meiotic cells. In mouse oocytes, there is significant Aurora kinase A (AURKA) compensatory abilities when the other Aurora kinase homologs are deleted. Whether the other homologs, AURKB or AURKC can compensate for loss of AURKA is not known. Using a conditional mouse oocyte knockout model, we demonstrate that this compensation is not reciprocal because female oocyte-specific knockout mice are sterile and their oocytes fail to complete meiosis I. In determining the AURKA-specific functions, we demonstrate that its first meiotic requirement is to activate Polo-like kinase 1 at microtubule organizing centers (MTOCs; meiotic spindle poles). This activation induces fragmentation of the MTOCs, a step essential for building a bipolar spindle. The next step that requires AURKA is building the liquid-like spindle domain that involves TACC3. Finally, we find that AURKA is also required for anaphase I onset to trigger cohesin cleavage in an APC/C independent manner. We conclude that AURKA has multiple functions essential to completing MI that are distinct from AURKB and AURKC.

scholarly journals Aurora kinase A is essential for meiosis in mouse oocytes

PLoS Genetics ◽  
2021 ◽  
Vol 17 (4) ◽  
pp. e1009327
Author(s):  
Cecilia S. Blengini ◽  
Patricia Ibrahimian ◽  
Michaela Vaskovicova ◽  
David Drutovic ◽  
Petr Solc ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Aurora Kinase ◽  
The Other ◽  
Meiotic Spindle ◽  
Multiple Functions ◽  
Aurora Kinase A ◽  
Mouse Oocytes ◽  
Spindle Poles ◽  
Knockout Model ◽  
Kinase A

The Aurora protein kinases are well-established regulators of spindle building and chromosome segregation in mitotic and meiotic cells. In mouse oocytes, there is significant Aurora kinase A (AURKA) compensatory abilities when the other Aurora kinase homologs are deleted. Whether the other homologs, AURKB or AURKC can compensate for loss of AURKA is not known. Using a conditional mouse oocyte knockout model, we demonstrate that this compensation is not reciprocal because female oocyte-specific knockout mice are sterile, and their oocytes fail to complete meiosis I. In determining AURKA-specific functions, we demonstrate that its first meiotic requirement is to activate Polo-like kinase 1 at acentriolar microtubule organizing centers (aMTOCs; meiotic spindle poles). This activation induces fragmentation of the aMTOCs, a step essential for building a bipolar spindle. We also show that AURKA is required for regulating localization of TACC3, another protein required for spindle building. We conclude that AURKA has multiple functions essential to completing MI that are distinct from AURKB and AURKC.

scholarly journals Aurora Kinase A Drives MTOC Biogenesis but Does Not Trigger Resumption of Meiosis in Mouse Oocytes Matured In Vivo1

2012 ◽  
Vol 87 (4) ◽  
Author(s):  
Petr Solc ◽  
Vladimir Baran ◽  
Alexandra Mayer ◽  
Tereza Bohmova ◽  
Gabriela Panenkova-Havlova ◽  
...  
Keyword(s):  
Aurora Kinase ◽  
Aurora Kinase A ◽  
Mouse Oocytes ◽  
Kinase A

A phase I study of Aurora kinase A inhibitor LY3295668 erbumine as a single agent and in combination in patients with relapsed/refractory neuroblastoma.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. TPS10561-TPS10561
Author(s):  
Steven G. DuBois ◽  
Jill D. Kremer ◽  
Bram De Wilde ◽  
Chad Jacobsen ◽  
Isabelle Aerts ◽  
...  
Keyword(s):  
Drug Exposure ◽  
Single Agent ◽  
Unmet Need ◽  
Study Drug ◽  
Aurora Kinase ◽  
Mycn Amplification ◽  
Hematologic Toxicity ◽  
Dependent Manner ◽  
Aurora Kinase A ◽  
Kinase A

TPS10561 Background: Aurora kinase A (AurA) has been implicated in high-risk neuroblastoma, including roles stabilizing and increasing expression of MYCN protein. AurA impacts the function of MYCN in mediating transcription in a cell cycle dependent manner, suggesting that neuroblastoma and other MYC/MYCN-driven tumors may be sensitive to AurA inhibition. LY3295668 is a selective AurA inhibitor. The lack of AurB inhibitory activity is hypothesized to minimize on-target hematologic toxicity associated with AurB inhibition. The molecule’s selectivity is intended to allow for continuous dosing at exposures associated with > 90% target inhibition at trough. In an analysis of LY3295668 antiproliferative effects in 560 cancer cell lines, neuroblastoma was among the most sensitive histologies tested. This screen also separately evaluated genomic predictors of response to LY3295668, with MYC/ MYCN amplification identified as among the strongest predictors of sensitivity to this agent. LY3295668 is currently being evaluated in early phase adult trials. The current trial (J10-MC-JZHD) was uniquely designed to hasten time to first-in-child oncology development for a rare unmet need of relapsed/refractory neuroblastoma patients. Methods: Study J1O-MC-JZHD (NCT04106219) is a multicenter, dual collaboration (NANT and ITCC), randomized, open-label, Phase 1 study of oral LY3295668 in children with relapsed/refractory neuroblastoma. A rolling 6 design will be followed for dose escalation in both a monotherapy cohort and a combination cohort testing LY3295668 together with cyclophosphamide and topotecan. The starting monotherapy dose will be equivalent to 80% of the adult maximum tolerated dose. Key eligibility criteria include recurrent/refractory neuroblastoma not amenable to curative treatment, age 2-21 years, mandatory archival tissue submission, ability to swallow capsules, and adequate hematologic and organ function. LY3295668 is administered in capsule form orally BID continuously. Primary objectives include assessments of safety and tolerability of study drug to identify RP2D as monotherapy and combination, and assess antitumor activity. Secondary objectives include assessment of the pharmacokinetic profile as monotherapy and in combination, and assessment of the relationship between study drug exposure and efficacy. Following determination of the RP2Ds, an expansion phase will randomize patients to monotherapy or to the combination arm. Enrollment began 16 Dec 2019 and is ongoing. Clinical trial information: NCT04106219.

scholarly journals Aurora kinase A controls meiosis I progression in mouse oocytes

Cell Cycle ◽  
2008 ◽  
Vol 7 (15) ◽  
pp. 2368-2376 ◽  
Author(s):  
Adela Saskova ◽  
Petr Solc ◽  
Vladimir Baran ◽  
Michal Kubelka ◽  
Richard M. Schultz ◽  
...  
Keyword(s):  
Aurora Kinase ◽  
Aurora Kinase A ◽  
Mouse Oocytes ◽  
Kinase A ◽  
Meiosis I

scholarly journals Aurora kinase A activates the vacuolar H+-ATPase (V-ATPase) in kidney carcinoma cells

2016 ◽  
Vol 310 (11) ◽  
pp. F1216-F1228 ◽  
Author(s):  
Mohammad M. Al-bataineh ◽  
Rodrigo Alzamora ◽  
Kazuhiro Ohmi ◽  
Pei-Yin Ho ◽  
Allison L. Marciszyn ◽  
...  
Keyword(s):  
Consensus Sequence ◽  
Aurora Kinase ◽  
Carcinoma Cells ◽  
Anacardic Acid ◽  
Transport Systems ◽  
Dependent Manner ◽  
Aurora Kinase A ◽  
Kidney Carcinoma ◽  
A Subunit ◽  
Kinase A

Extracellular proton-secreting transport systems that contribute to extracellular pH include the vacuolar H+-ATPase (V-ATPase). This pump, which mediates ATP-driven transport of H+across membranes, is involved in metastasis. We previously showed (Alzamora R, Thali RF, Gong F, Smolak C, Li H, Baty CJ, Bertrand CA, Auchli Y, Brunisholz RA, Neumann D, Hallows KR, Pastor-Soler NM. J Biol Chem 285: 24676–24685, 2010) that V-ATPase A subunit phosphorylation at Ser-175 is important for PKA-induced V-ATPase activity at the membrane of kidney intercalated cells. However, Ser-175 is also located within a larger phosphorylation consensus sequence for Aurora kinases, which are known to phosphorylate proteins that contribute to the pathogenesis of metastatic carcinomas. We thus hypothesized that Aurora kinase A (AURKA), overexpressed in aggressive carcinomas, regulates the V-ATPase in human kidney carcinoma cells (Caki-2) via Ser-175 phosphorylation. We found that AURKA is abnormally expressed in Caki-2 cells, where it binds the V-ATPase A subunit in an AURKA phosphorylation-dependent manner. Treatment with the AURKA activator anacardic acid increased V-ATPase expression and activity at the plasma membrane of Caki-2 cells. In addition, AURKA phosphorylates the V-ATPase A subunit at Ser-175 in vitro and in Caki-2 cells. Immunolabeling revealed that anacardic acid induced marked membrane accumulation of the V-ATPase A subunit in transfected Caki-2 cells. However, anacardic acid failed to induce membrane accumulation of a phosphorylation-deficient Ser-175-to-Ala (S175A) A subunit mutant. Finally, S175A-expressing cells had decreased migration in a wound-healing assay compared with cells expressing wild-type or a phospho-mimetic Ser-175-to-Asp (S175D) mutant A subunit. We conclude that AURKA activates the V-ATPase in kidney carcinoma cells via phosphorylation of Ser-175 in the V-ATPase A subunit. This regulation contributes to kidney carcinoma V-ATPase-mediated extracellular acidification and cell migration.

scholarly journals Dynamic Changes in pStat3 Are Involved in Meiotic Spindle Assembly in Mouse Oocytes

2020 ◽  
Vol 21 (4) ◽  
pp. 1220
Author(s):  
Seiki Haraguchi ◽  
Mitsumi Ikeda ◽  
Satoshi Akagi ◽  
Yuji Hirao
Keyword(s):  
Oocyte Maturation ◽  
Spindle Assembly ◽  
Meiotic Spindle ◽  
Dual Function ◽  
Dependent Manner ◽  
Vitro Fertilization ◽  
Mouse Oocytes ◽  
Spindle Poles ◽  
Transcription 3

The signal transducer and activator of transcription 3 (Stat3) is activated upon phosphorylation at Y705 (pStat3) and serves the dual function of signal transduction and transcription activation. Our previous study suggested that pStat3 is functional during oocyte maturation when transcription is silenced. Therefore, we speculated that pStat3 serves other functions. Immunocytochemical analysis revealed that pStat3 emerges at microtubule asters and spindle and is subsequently localized at the spindle poles along with pericentrin during mouse oocyte maturation. Both Stat3 and pStat3 proteins were detected in conditionally knocked out Stat3−/− mouse oocytes. pStat3 localization was the same in Stat3+/+ and Stat3−/− oocytes, and oocyte maturation proceeded normally, suggesting that pStat3 was still functional. Furthermore, the treatment of oocytes with the Stat3-specific inhibitors stattic and BP-1-102 or anti-pStat3 antibody led to significantly abnormal spindle assembly and chromosome mislocation in a dose-dependent manner, and pStat3 was either absent or improperly localized in these oocytes. Moreover, the development of pre-implantation stage embryos derived from inhibitor-treated oocytes was significantly hampered following in vitro fertilization. These findings indicate a novel function of pStat3 in spindle assembly.

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