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斯皮克斯黄齿豚鼠(Galea spixii Wagler,1831)卵巢腔前卵泡群体的特征分析与冷冻保存

Characterisation and cryopreservation of the ovarian preantral follicle population from Spix's yellow-toothed cavies (Galea spixii Wagler, 1831).

作者信息

Praxedes Érica C G, Lima Gabriela L, Silva Andréia M, Apolinário Carlos A C, Bezerra José A B, Souza Ana L P, Oliveira Moacir F, Rodrigues Ana P R, Silva Alexandre R

机构信息

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido (UFERSA), BR 110, Km 47, Costa e Silva, 59625-900, Mossoró, RN, Brazil.

Laboratory of Manipulation of Oocytes Enclosed in Preantral Follicles (LAMOFOPA), Veterinary Faculty, Universidade Estadual do Ceará (UECE), Paranjana Avenue, 1700, Itaperi, 60740-000, Fortaleza, CE, Brazil.

出版信息

Reprod Fertil Dev. 2017 Mar;29(3):594-602. doi: 10.1071/RD15249.

Abstract

The aim of the present study was to characterise the ovarian preantral follicle (PF) population and to establish a solid surface vitrification (SSV) process using dimethyl sulfoxide (DMSO) as a cryoprotectant for preservation of ovarian tissue from yellow-toothed cavies (Galea spixii). Ovaries were fixed for PF population analysis or were subjected to the SSV process. The mean (± s.e.m.) PF population per ovarian pair was estimated to be 416.0±342.8. There were 140.0±56.0 (63.4%) and 125.0±58.0 (64.0%) primary follicles on the right and left ovaries, respectively. The proportion of this follicle category was significantly greater than that of other follicle categories (P<0.05). The diameter of follicles (123.7±18.3µm), oocytes (50.1±5.0µm) and nuclei (14.27±2.01µm) was larger for secondary ones when compared with other PFs categories. Most PFs were morphologically normal (94.6%), with light microscopy identifying only a few atretic follicles (5.4%). After SSV, there was a reduction in the proportion of morphologically normal PFs compared with the non-vitrified group (69.5% vs 91.2%, respectively). Transmission electron microscopy revealed preservation of oocytes and granulosa cell membranes and the morphological aspect of follicles; the primary change observed in some vitrified PFs was the presence of vacuoles in the oocytes and granulosa cells cytoplasm and turgid mitochondria. In conclusion, the present study provides an estimative and characterization for the PF population in ovaries of G. spixii. Moreover, we report its PFs cryopreservation using an SSV process.

摘要

本研究的目的是对卵巢窦前卵泡(PF)群体进行特征描述,并建立一种使用二甲基亚砜(DMSO)作为冷冻保护剂的实体表面玻璃化(SSV)方法,用于保存黄齿豚鼠(Galea spixii)的卵巢组织。卵巢被固定用于PF群体分析或进行SSV处理。估计每对卵巢的平均(±标准误)PF群体数量为416.0±342.8。右侧和左侧卵巢分别有140.0±56.0(63.4%)和125.0±58.0(64.0%)个初级卵泡。该卵泡类别的比例显著高于其他卵泡类别(P<0.05)。与其他PF类别相比,次级卵泡的卵泡直径(123.7±18.3µm)、卵母细胞直径(50.1±5.0µm)和细胞核直径(14.27±2.01µm)更大。大多数PF在形态上是正常的(94.6%),光学显微镜仅识别出少数闭锁卵泡(5.4%)。SSV处理后,与未玻璃化组相比,形态正常的PF比例有所降低(分别为69.5%和91.2%)。透射电子显微镜显示卵母细胞和颗粒细胞膜以及卵泡的形态得以保存;在一些玻璃化的PF中观察到的主要变化是卵母细胞和颗粒细胞细胞质中存在空泡以及线粒体肿胀。总之,本研究提供了对黄齿豚鼠卵巢中PF群体的估计和特征描述。此外,我们报告了使用SSV方法对其PF进行冷冻保存的情况。

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