Oocyte meiotic-stage-specific differences in spindle depolymerization in response to temperature changes monitored with polarized field microscopy and immunocytochemistry

Gomes, Cristina Marques; Merlini, M.; Konheim, Jeremy; Serafini, P.; Motta, E.L.A.; Baracat, Edmund C.; Smith, Gary D.

doi:10.1016/j.fertnstert.2011.12.018

Cited by 15 publications

(10 citation statements)

References 31 publications

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“…However, the inability of polarized light microscopy to distinguish between spindles with normal (bipolar) and highly disarranged conformation and to predict the degree of microtubule polymerization in metaphase II (MII) spindles of frozen/thawed oocytes make it an inefficient method to assess the MII spindle, especially after cryopreservation [57]. Recently, Gomes et al [58] used polarized field microscopy, a noninvasive imaging method, and immunocytochemistry to compare the polymerization status of mouse oocyte spindles at various stages of meiosis, metaphase I (MI), telophase I (TI), and MII exposed to various temperatures (37°C, room temperature, 4°C, and vitrification) for 0, 10, 30, and 60 min. They found that the temperature-and time-dependent differences in the depolymerization/ repolymerization equilibrium of oocyte spindles are related to the meiotic stage, in which TI shows less depolymerization at room temperature, 4°C, and after vitrification and warming than that of spindles in MI and MII oocytes.…”

Section: Cytoskeletonmentioning

confidence: 99%

“…However, there is disappearance and reappearance of meiotic spindles during MII after vitrification and slow freezing [35,6163], which depends on the time interval after thawing, methods of freezing and thawing, and the species [55,59,60,64]. It has also been proposed that temperature-induced oocyte microtubule depolymerization may be dependent on the nuclear maturation state of oocytes [58,65]. A potential strategy to avoid spindle depolymerization is cryopreservation of oocytes at the germinal vesicle (GV) stage.…”

Section: Cytoskeletonmentioning

confidence: 99%

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Cryopreservation of mammalian oocytes and embryos: current problems and future perspectives

Moussa

Shen

Zhang

et al. 2014

Sci. China Life Sci.

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Cryopreservation techniques for mammalian oocytes and embryos have rapidly progressed during the past two decades, emphasizing their importance in various assisted reproductive technologies. Pregnancies and live births resulting from cryopreserved oocytes and embryos of several species including humans have provided proof of principle and led to the adoption of cryopreservation as an integral part of clinical in vitro fertilization. Considerable progress has been achieved in the development and application of the cryopreservation of mammalian oocytes and embryos, including preservation of the reproductive potential of patients who may become infertile, establishment of cryopreserved oocyte banks, and transport of oocytes and embryos internationally. However, the success rates are still far lower than those obtained with fresh oocytes and embryos, and there are still obstacles that need to be overcome. In this review, we address the major obstacles in the development of effective cryopreservation techniques. Such knowledge may help to eliminate these hurdles by revealing which aspects need improvement. Furthermore, this information may encourage further research by cryobiologists and increase the practical use of cryopreservation as a major part of assisted reproductive technologies for both humans and animal species. cryopreservation, mammals, oocytes, embryos, cryoinjuries Citation:Moussa M, Shu J, Zhang XH, Zeng FY. Cryopreservation of mammalian oocytes and embryos: current problems and future perspectives. Sci China Life Sci, 2014, 57: 903 -914,

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Section: Cytoskeletonmentioning

confidence: 99%

Section: Cytoskeletonmentioning

confidence: 99%

Cryopreservation of mammalian oocytes and embryos: current problems and future perspectives

Moussa

Shen

Zhang

et al. 2014

Sci. China Life Sci.

View full text Add to dashboard Cite

show abstract

“…Thus, it is likely that inclusion of D 2 O and Taxol in the fixative may have caused aberrant polymerization of microtubules during fixation in the previous study [26]. Disappearance of the meiotic spindle during cryopreservation is also consistent with the cold-labile nature of microtubules [43,50], and exposure even to room temperature causes abnormal spindle configuration in mouse and human oocytes [9,14,42]. Importantly, however, the present study showed that disappearance of spindle microtubules in the vitrification solutions are not solely due to exposure to room temperature, as incubation of oocytes in FHM at the ambient temperature for the duration comparable to the V1 to V3 treatments or even for 2 h did not significantly diminish the meiotic spindle.…”

Section: Discussionmentioning

confidence: 62%

“…To examine whether D 2 O and Taxol can cause microtubule polymerization after complete disappearance of the meiotic spindle, we conducted the following experiment. First, MII oocytes were incubated in FHM on ice for 45 min to depolymerize spindle microtubules, as microtubules are highly susceptible to cold temperatures [9,14,[41][42][43]. This treatment completely depolymerized spindle microtubules in all the oocytes examined (n ¼ 23), as shown by immunocytochemistry for b-Tubulin (Fig.…”

Section: Formation Of Spindle-like Microtubule Bundles Induced By Deumentioning

confidence: 87%

“…Some observations were based on polarized light microscopy, which enables visualization of highly ordered molecules, namely, bundles of spindle microtubules, in live oocytes [31,[43][44][45]. Other observations were made by immunostaining, which allows detection of spindle microtubules in much higher resolutions, although it requires fixation of specimens [2,[9][10][11]14].…”

Section: Discussionmentioning

confidence: 99%

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Impact of Vitrification on the Meiotic Spindle and Components of the Microtubule-Organizing Center in Mouse Mature Oocytes1

Tamura

Huang

Marikawa

2013

Biology of Reproduction

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Cryopreservation of oocytes is becoming a valuable method for fertility preservation in women. However, various unphysiological alterations occur in the oocyte during the course of cryopreservation, one of which is the disappearance of the meiotic spindle. Fortunately, the meiotic spindle does regenerate after thawing the frozen oocytes, which enables completion of meiosis and further development after fertilization. Nonetheless, the mechanistic understanding of the meiotic spindle regeneration after cryopreservation is still scarce. Here, to gain insight into the mechanisms of the spindle disappearance and regeneration, we examined the status of spindle microtubules as well as the key components of the microtubule-organizing center (MTOC), specifically gamma-Tubulin, NEDD1, and Pericentrin, in mature (metaphase II) mouse oocytes at different steps of vitrification, a major cryopreservation technique. We found that the configuration of the spindle microtubules dynamically changed during the process of vitrification and that spindle regeneration was preceded by excessive microtubule polymerization, followed by reduction into the normal size and shape. Also, all three MTOC components exhibited disappearance and reappearance during the vitrification process, although Pericentrin appeared to regenerate in earlier steps compared to the other components. Furthermore, we found that the localization of the MTOC components to the spindle poles persisted even after depolymerization of spindle microtubules, suggesting that the MTOC components are impacted by vitrification independently from the integrity of the microtubules. The present study would set the stage for future investigations on the molecular mechanisms of the meiotic spindle regeneration, which may contribute to further improving protocols for oocyte cryopreservation.

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Temperature Control in IVF Culture System

Li,

Gao

2024

Quality Management in the Assisted Reproduction Laboratory

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Oocyte meiotic-stage-specific differences in spindle depolymerization in response to temperature changes monitored with polarized field microscopy and immunocytochemistry

Cited by 15 publications

References 31 publications

Cryopreservation of mammalian oocytes and embryos: current problems and future perspectives

Cryopreservation of mammalian oocytes and embryos: current problems and future perspectives

Impact of Vitrification on the Meiotic Spindle and Components of the Microtubule-Organizing Center in Mouse Mature Oocytes1

Temperature Control in IVF Culture System

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