Effects of Chilling on Structural Aspects of Early Preantral Mouse Follicles1
Effects of Chilling on Structural Aspects of Early Preantral Mouse Follicles1
复制标题
低温对小鼠早期腔前卵泡结构的影响1
DOI：
10.1095/biolreprod.103.020933
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
J. Smitz
中科院分区：
文献类型：
--
作者：
L. Vanhoutte;R. Cortvrindt;D. Nogueira;J. Smitz
摘要冷冻损伤是影响配子低温保存效果的主要因素之一。本研究的目的是确定潜在的冷冻诱导的几个结构方面的早期腔前小鼠卵泡的损害。将机械分离的完整早期腔前卵泡（3b-4型）暴露于0°C下1、5、10或30 min。对照和冷冻卵泡在微管蛋白、F-肌动蛋白和染色质染色后通过共聚焦显微镜和电子显微镜进行分析。冷冻仅1分钟就足以引起卵母细胞和周围颗粒细胞层中微管的解聚，这一点可以通过抗微管蛋白标记后荧光强度的大幅降低来证明。较长时间的冷却导致卵泡封闭的卵母细胞中微管组织的改变。这些变化包括间期微管的丢失，伴随着核周或皮质微管星状体的形成，有时微管完全消失。微管修饰的程度与冷冻时间有关，但在37°C复温1 h后完全可逆。冷冻对主要位于卵母细胞皮层和跨带投射的含肌动蛋白的元素只有轻微的影响。超微结构分析证实，卵母细胞-体细胞相互作用的存在。对卵泡封闭的卵母细胞内的染色质构型没有影响。这些结果表明，小鼠卵泡对直接冷损伤具有相对耐受性，因此能够承受传统冷冻保存过程中的冷却-升温步骤。
Abstract Chilling injury is one of the major limiting factors for achieving optimal cryopreservation of gametes. This study aimed to determine potential chilling-induced damage on several structural aspects of early preantral mouse follicles. Mechanically isolated intact early preantral follicles (type 3b-4) were exposed to 0°C for 1, 5, 10, or 30 min. Control and chilled follicles were analyzed by confocal microscopy after staining for tubulin, F-actin, and chromatin, and by electron microscopy. Chilling for only 1 min was sufficient to cause depolymerization of microtubules in the oocyte and the surrounding granulosa cell layer as evidenced by a substantial decrease in fluorescence intensity after antitubulin labeling. Cooling for longer periods caused alterations in microtubule organization in the follicle-enclosed oocyte. These alterations included the loss of interphase microtubules, concomitant with the formation of perinuclear or cortical microtubule asters and sometimes a complete disappearance of microtubules. The extent of microtubule modification was related to the time of chilling, but was fully reversible after rewarming follicles at 37°C for 1 h. Chilling had only minor effects on the actin-containing elements located predominantly in the oocyte cortex and the transzonal projections. Ultrastructural analysis confirmed that oocyte-somatic cell interactions were present. There was no influence on the chromatin configuration within the follicle-enclosed oocyte. These results indicate that mouse follicles are relatively tolerant to direct chilling injury and, as a consequence, are able to withstand the cooling-warming steps during conventional cryopreservation procedures.
影响因子：
2.7
作者：
WICKRAMASINGHE, D;EBERT, KM;ALBERTINI, DF
通讯作者：
ALBERTINI, DF
DOI：
10.1016/0165-1110(92)90032-5
发表时间：
1992-12-01
期刊：
MUTATION RESEARCH
影响因子：
--
作者：
ALBERTINI, DF
通讯作者：
ALBERTINI, DF
小鼠卵母细胞减数分裂过程中的中心体和微管动力学。
影响因子：
4
作者：
Messinger,SM;Albertini,DF
通讯作者：
Albertini,DF